English Hb9210 PDF

Premier Hb9210TM Boronate Affinity | 09-00-0001 | 11-00-0001 | Revision 23
HAEMOGLOBINS SYSTEMS DIVISION
Operator’s Manual

IVD. FOR IN VITRO DIAGNOSTIC USE ONLY
CAUTION: Before using the Premier Hb9210TM, please become familiar with all the
instructions in this manual.
Serial Number
_________________________________________
Proprietary Information
Software programs are protected by copyright. All rights are reserved. This software was
developed solely for use with intended equipment and for “in vitro” diagnostic applications as
specified in the operating instructions. All operating instructions must be followed. Copying or
other reproduction of the program is prohibited without the prior written consent of Trinity
Biotech.
Applications for Assay of variant haemoglobins, glycated haemoglobins and other proteins are
covered by one or more of the following US Patents:
5,843,788 5,719,053 6,020,203 5,801,053 9,164,115
Copyright © 2017
Contact addresses:
Support:
US/Canada/Puerto Rico Customers: Trinity Biotech USA 2823 Girts Road Jamestown, NY 14701 Tel:
1-800-325-3424 www.trinitybiotech.com [email protected]
International Customers: Trinity Biotech IDA Business Park Bray, Co. Wicklow, Ireland Tel: + 353 1 276 9898 Fax:
+ 353 1 276 9888 www.trinitybiotech.com [email protected]

1. Site Preparation
2. Introduction & System Description
3. Principles of Operation
4. Preparation for Operation
5. Hardware Components
6. Software & Operation
7. Results & Interpretation
8. Operator Maintenance
9. Operator Troubleshooting
10. Data Transmission
TLA Appendix

Expanded Table of Contents
Chapter 1: Site Preparation
1.1 Introduction 1 1.2 System Description 1 1.3 System Requirements 1 1.3.1 Environmental
Requirements 1 1.3.2 Electrical Requirements 1 1.4 Installation 2 1.4.1 Components Transport and
Storage Prior to Installation 2 1.4.2 Safe Lifting Technique 3 1.4.3 Instrument Installation Warnings 3 1.5
Important Symbols 4 1.6 Inspection 5 1.7 Important Safety Instructions 5
Chapter 2: Introduction & System Description

2.1 Introduction 1 2.2 Features 2 2.3 Specifications 3 2.4 CPU Specifications 3 2.5 Intended Use 4 2.6
Premier Hb9210TM Certification 4 2.7 510k Listing 4 2.8 CE Mark 4
Chapter3: Principles of Operation

3.1 Clinical Significance of HbA1c 1 3.2 Principles of Analysis 1 3.3 Merits of the Assay 3 3.3.1
Boronate Affinity HbA1c Assay 3 3.3.2 Premier Hb9210TM 3 3.3.3 Glycated Haemoglobin Reporting
Units 3 3.3.4 Expected Values 4 3.3.5 HbA1c Accuracy 4 3.3.6 HbA1c Linearity/Measurable Range 5
3.3.7 HbA1c Analytical Specificity 5 3.3.8 HbA1c Carryover 6 3.3.9 HbA1c Matrix Effects – Collection
Methods and Anticoagulants 7 3.3.10 HbA1c Matrix Effects – Frozen and non-Frozen Samples 7 3.3.11
HbA1c Precision 8 3.4 Chapter References 10 Appendix A - Multicentre Evaluation of the Premier
Hb9210TM HbA1c Analyser 11

Chapter 4: Preparation for Operation
4.1 Test components 1 4.2 Reagent Use and Storage 3 4.3 Preparation of Haemoglobin HbA1c
Calibrators and Controls 4 4.3.1 Trinity Biotech Haemoglobin HbA1c Calibrators 4 4.3.2 Trinity Biotech
Haemoglobin HbA1c Controls 4 4.4 Column Use and Storage 4 4.4.1 General Column Information 4
4.4.2 Column Storage 5 4.4.3 Column Installation 5 4.4.4 Column Use 5 4.4.5 Summary 6 4.5 Specimen
Requirements 6 4.5.1 Collection 6 4.5.2 Patient Sample Storage Stability 6 4.5.3 Premier Hb9210TM
Sample Storage Stability Study 6 4.5.4 Anaemic, Normal, pRBC and Washed RBC Sample Dilutions 7
4.5.5 Sample Racks 8 4.5.6 Common Errors with Sample Tubes 10 4.6 Limitations of the Assay 10
Chapter 5: Hardware Components

5.1 Introduction 1 5.1.1 A note on Using Manufacturer-Approved Replacement Parts 1 5.2 Autosampler
System 2 5.2.1 Sample Transport Module Description (09-30-1300S) 2 5.2.2 Probe Module Description
(09-30-1620S) 3 5.2.3 Injection Module Description (09-30-1900S) 4 5.3 Pump Module (09-30-2000S) 4
5.3.1 System Description 4 5.4 Oven Module (09-30-1800S) 5 5.4.1 System Description 5 5.4.2
Performance Specifications 5 5.5 Detector Module (09-30-1700S) 5 5.5.1 Detector Description 5 5.6 CPU
Module (09-30-1500S) 6 5.6.1 Data Handling Description 6 5.6.2 Physical Characteristics 6 5.7 Barcode
Readers 7 5.7.1 Manual Barcode Reader Gun 7 5.7.2 Automatic Barcode Reader Assembly (on Sample
Transport) 8 5.7.3 Barcode Sticker Application 8 5.8 Automatic Debubblers & Waste Pump 9 5.8.1
Automatic Debubblers 9 5.8.2 Waste Pump 10 5.9 Decontamination Protocol for Service and End of Life
Disposal 11

Chapter 6: Software & Operation
6.1 Introduction & Start Up 1 6.1.1 Definitions 1 6.2 Installation Screens 2 6.3 Preparation for Patient
Sample Runs 13 6.3.1 Activate System 13 6.3.2 System Calibration 15 6.3.3 Baseline Calibration 17 6.3.4
Load Racks 19 6.3.5 Patient Sample Run 19 6.3.6 Automated operator Assistant 21 6.4 Deactivate
System/Exit Application 22 6.5 Other Menus & Software Tools 24 6.5.1 Operation Menu 24 6.5.2 Edit
Menu 27 6.5.3 View Menu 28 6.5.4 Devices Menu 33 6.5.5 Help Menu 38 6.5.6 Exit Application 39 6.6
Directory Structure 40 6.7 Copying from the GHbArch Folder 41
Chapter 7: Results & Interpretation

7.1 Overview 1 7.2 Auto-Verification 1 7.2.1 Auto-Verification Features 2 7.2.2 Auto-Verification
Conclusions 9 7.3 QC Recommendations 10 7.4 Report Interpretation 11 7.4.1 On-Screen Report 11 7.4.2
Printed Report 11 7.4.3 Interpretation of Report 12 7.5 Other Reports 13 7.5.1 Header Page 13 7.5.2
Batch Summary Report (BSR) 16 7.6 Chapter References 17

Chapter 8: Operator Maintenance
8.1 Introduction, Instrument & Component Overview 1 8.1.1 Daily Maintenance 2 8.1.2 Weekly
Maintenance 3 8.1.3 Periodic Maintenance 3 8.1.4 Preventative Maintenance 3 8.2 Changing the
Reagents 4 8.2.1 Priming the Pumps 9 8.2.2 Priming the Piston Rinsing Chamber with Purified Water 9
8.2.3 Priming the Dilutor (Premier Hb9210TM DIL Reagent) 10 8.2.4 Priming the Active Rinse Station
(ARS) 10 8.3 Changing the Column 11 8.3.1 Changing the Frits 16 8.4 Detector Maintenance 20 8.4.1
Removing Air Bubbles from the Flow Cell 20 8.5 Pump Maintenance 21 8.6 Sample Transport, Syringe
Module and Probe Module Maintenance 21
Chapter 9: Operator Troubleshooting

9.1 General Troubleshooting 1 9.1.1 Aligning the Touch Screen Monitor 1 9.1.2 Priming the Manual
Debubblers 3 9.2 Hardware Error Codes 5 9.2.1 Data Handling System 5 9.2.2 Autosampler System 7
9.2.3 Pump Module 11 9.2.4 Oven Module 12 9.2.5 Detector Module 12 9.2.6 Waste Pump 13 9.3 Peak
Analysis & Results Codes 14 9.3.1 Patient Sample Peak Analysis & Results Codes 14 9.3.2 Calibration
Run Peak Analysis & Results Codes 22 9.4 General Chromatography Troubleshooting 31 9.4.1 Normal
Chromatography 31 9.4.2 General Recommendations 32 9.4.3 Hb Variants & Other Potentially
Interfering Substances 32 9.4.4 Air In Pumps 33 9.4.5 Short Sample 33 9.5 Miscellaneous Operational
Suggestions 34 9.5.1 Proper Handling of Biohazardous Waste 34 9.6 Contacting Technical Support 35

Chapter 10: Data Transmission
10.1 Premier Hb9210TM Data Transmission using ASTM E1394-97 Protocol 1 10.1.1 Introduction 1
10.1.2 General 1 10.1.3 Physical Connection & Communication 1 10.2 ASTM E1394-97 Field Usage 2
10.2.1 Header Record 2 10.2.2 Patient Record 3 10.2.3 Order Record 3 10.2.4 Result Number 1 Record ...
Result Number 3 Record 5 10.2.5 Message Terminator 10 10.2.6 Communication to LIS Example 10
10.2.7 Bi-directional Communication between the Premier Hb9210TM and the LIS 10 10.3 Premier
Hb9210TM Data Transmission using Data Dump 11 10.3.1 Introduction 11 10.3.2 General 11 10.3.3
Physical Connection & Communication 11 10.3.4 Data Field Usage 11 10.3.5 Premier Hb9210TM
Communication to LIS Example 12 10.3.6 Uni-directional Communication Between Premier Hb9210TM
and the LIS 12
TLA Appendix
TLA 1 System Overview 1 TLA 2 System Operation 3 TLA 3 TLA LIS Protocol 9

PREMIER Hb9210TM Site
Preparation
CHAPTER CONTENTS
✓ System Requirements ✓ Installation ✓ Safety Precautions

Chapter 1 – Site Preparation
1.1 Introduction
This section defines environmental, physical and electrical requirements of the Premier Hb9210TM
HPLC instrument.
1.2 System Description

The system consists of Premier Hb9210TM Liquid Chromatograph containing integrated HPLC, sample
preparation/transport, and computer workstation.
Instrument Dimensions
Height: 74 cm (29”) Width: 54 cm (21”) Depth: 66 cm (26”) Instrument Weight 62 kg (137 pounds)
1.3 System Requirements

1.3.1 Environmental Requirements
Temperature 10 – 40°C (50-104°F) Humidity 10 – 90% (non-condensing)
Required Bench Space
76 cm (30”) of bench space that is at least 76 cm (30”) deep. Must be capable of supporting 75 Kg
(165lbs) Storage Condition 2 – 40°C Storage Humidity 10-90% (non-condensing)
1.3.2 Electrical Requirements
Line Voltage
110/120 VAC +5%, -10% 220/240 VAC +5%, -10%
1 | Chapter 1 – Site Preparation Rev. 23
Only an authorized Trinity Biotech representative should perform the installation of the Premier
Hb9210TM system. Installation by any other person(s) will void the system warranty. Moving the
instrument within or external to the laboratory also requires the presence of an authorized Trinity Biotech
Representative.

Line Frequency 50 – 60 Hz
Line Disturbance
Surges and sags must not exceed ± 20% of normal line voltage. Voltage must return to normal within 15
msec, line transients >10μsec pulse width and >50% of normal rated may produce instrument
malfunctions. Line Voltage 100 – 240 VAC Power Connection 3-prong ground outlet Power
Consumption 250 watts maximum
Fuse Rating
3.0 Amp, 250VAC, Qty: 2 Type: GMA
1.4 Installation
Installation of the Premier Hb9210TM HPLC instrument is performed only by authorized Trinity Biotech
representatives, for the most up-to-date installation instructions please contact Trinity Biotech or your
local distribution partner. Unauthorized installation may invalidate the instrument warranty.
1.4.1 Component Transport and Storage Prior to Installation
1. Components and system cabinet should be transported in their individual packing containers with
custom
foam inserts present.
2. Palletized entire system can only be stacked 1 unit high.
3. Follow all symbols on packaging, as shown in Figure 2 below.
4. Instrument components need to be shipped and stored in a relatively cool and dry location, 10 - 40°C
(50-
104°F) and 10 - 90% relative humidity (non-condensing).
The power line assigned to this instrument should not be shared with any other equipment due to potential
interference and power problems. To protect the operator, the instrument panels and cabinet must be
grounded. Verify the ground before installing the System.

1.4.2 Safe Lifting Technique
When lifting the Premier Hb9210TM instrument, it is important to practice safe lifting techniques. Safe
lifting instructions are as follows:
1. Stand close to the instrument component.
2. Bend your knees when lifting; do not use your back.
3. Let your legs do the lifting while keeping your back straight.
4. Practice team lifting (get somebody to help you) if necessary.
✓
x
Figure 1: Proper and Improper Lifting Technique
1.4.3 Instrument Installation Warnings
1. The system waste tubing needs to be checked for proper and correct drainage to a clearly marked
biohazard container or sanitary drain system. Incorrect drainage of the system solution(s) could result in a
biohazard to personnel.
2. Disposal of system waste must be in accordance with local regulations.

1.5 Important Symbols
1. 2. 3. 4. 5. 6.
7. 8. 9. 10. 11. 12.
13. 14. 15. 16. 17. 18.
Figure 2: Important Symbols
1. Biohazard 2. Grounding (Earth Grounding) 3. Grounding (Earth Grounding) 4. Warning: Surface Hot
5. Main Power On/Off 6. Transport with care 7. Components fragile 8. Do not get wet 9. Use no hooks
10. Do not expose components to strong magnetic fields 11. Box and components recyclable 12. Do not
stack more than “X” number of units high 13. This end up 14. Pinch point 15. Press/Crush point 16.
Dispose electrical components in accordance with
local regulations 17. Read Instructions Carefully Before Use 18. Device Interface Module
Figure 3 below shows other important symbols listed on the rear of the instrument.
Figure 3: Example of Rear Panel Label

1.6 Inspection
Inspect all cartons delivered. If you find signs of external damage, call Trinity Biotech. Do not unpack
cartons until your installation representative has arrived at your site.
1.7 Important Safety Instructions

✓ Check that the voltage setting matches the supply voltage. ✓ Connection to mains supply: Where
protective grounding is required, plug the equipment into a supply outlet
which has an earth connection. ✓ Do not place the equipment in liquid, nor put it where it could fall
into liquid. ✓ If the equipment becomes wet, unplug it before touching it. ✓ Use the equipment only for
the purpose described in the instructions for use. ✓ Do not use accessories which are not supplied or
recommended by the manufacturer – these will void
warranty, claims and registrations. ✓ Do not use the equipment if it is not working properly, or if it
has suffered any damage.
Misuse of electrical equipment can cause electrocution, burns, fire and other hazards. Basic safety
precautions should always be taken, including all of the following listed below. Read before using the
equipment.

PREMIER Hb9210TM
CHA
PTER CONTENTS
Introduction & System
✓ Introduction ✓ Features ✓ Specifications Description
✓ Intended Use

Chapter 2 – Introduction & System Description
2.1 Introduction
The Premier Hb9210TM System consists of an integrated HPLC (2 Reagent Pumps, Injection Valve,
Oven and Detector), compact sample preparation system (Probe/Injection Port, Syringe Module and
Sample Transport) and the workstation (CPU and Touch Screen Interface) with the Premier Hb9210TM
Application Software.
Figure 1: The Premier Hb9210TM HPLC
The System consists of four main components used to fractionate and quantitate glycated haemoglobins:
1. The Premier Hb9210TM HPLC is composed of a dual high pressure pump system, column oven and
detector.
1 | Chapter 2 – Introduction & System Description Rev.23

2. The integrated autosampler/sample transport features a 210 sample capacity to start a single batch run
with the ability to continuously unload analyzed samples and load new samples. The additional STAT
sample capability allows for the analysis of an additional individual sample during batch analysis.
3. The system is interfaced to a personal computer for system control and data storage.
4. The Windows 7 or XPTM embedded software provides real-time chromatography and system
condition monitoring. Patient sample identification is automated during the sampling process via barcode
module in sample transport area. A barcode gun is supplied for hand-scanning of consumable barcodes,
package inserts, as well as easy inputting of Calibrator and Control material set points.
The fractionation and quantitation of glycated haemoglobin is accomplished using the principles of
boronate affinity high performance liquid chromatography.
2.2 Features
1. Full walk-away capability
2. Auto-verification of patient sample results
3. Windows 7 or XPTM embedded, touch screen, icon-driven, user-friendly software control
4. Small sample volume (0.5mL minimum haemolysate or mixed whole blood)
5. Ability to start a batch of up to 210 samples with the ability to continuously unload analyzed samples
and load
new samples
6. STAT sample capability
7. Automatic sampling and injection system
8. Integrated barcoding-while-sampling scanning of patient sample identification information
9. Individual sample reports with selectable units (mmol HbA1c/mol Hb (IFCC) & %HbA1c (NGSP)),
retention
times and area of peaks
10. Central processing unit (CPU) for automatic and unlimited storage of test results and archived test
results
11. Summary report of test results from each batch run – for on-screen review and/or printing
12. Ability to reprint batch summary reports and chromatograms from computer storage and archive
13. Real-time display allows monitoring of system operation
The software allows the operator to re-use the same sample rack during batch runs when running samples
without barcodes (identified as ‘noID’ samples in software). Please see Chapter 6, section 6.3.5 for more
details.

2.3 Specifications
1. Sample Whole blood, haemolysates or packed red blood cells.1
2. Elements of Analysis HbA1c
3. Principle Boronate affinity high performance liquid chromatography (HPLC)
4. Minimum readable division Area percent = 0.01%
5. Minimum sample requirement ✓ 10μL (whole blood to make 1:150 haemolysate dilution,
0.5mL minimum haemolysate volume) ✓ 5μL (packed red blood cells to make 1:300 dilution, 0.5mL
minimum haemolysate volume) ✓ 0.5mL (mixed whole blood to make direct injection) 6. Sample
capacity 210 samples
7. Throughput 48 samples per hour
8. Analytical Column Boronate bonded to a porous polymer support gel
9. Column temperature 55.0oC ± 0.2oC
10. Autosampler Integrated 210 sample transport
11. Autoinjector Delivers sample volume with <1% error with 5-μL injection volume
12. Detection LED wavelength detector, 413 + 2nm
2.4 CPU Specifications

1. Computer workstation comes pre-configured with Microsoft Windows 7 or XPTM Embedded
operating system. 2. CPU has unlimited storage capacity for test results and archived test results 3.
Hardware and/or software additions to the computer without consent of Trinity Biotech are not allowed.
Any
changes to the computer may interfere with validity of patient results and system integrity. 4. If
Operator’s require a printer to be used with the Premier Hb9210TM, the printer cable for the computer
must
be Windows 7 or XPTM and USB compatible.
1 Refer to section Chapter 4, Section 5 for information on Specimen Requirements

2.5 Intended Use
The intended use statement for the Premier Hb9210TM HbA1c system running the boronate affinity
HbA1c assay is listed below.
The Premier Hb9210 System is intended for the quantitative measurement of haemoglobin A1c
(HbA1c) in human capillary and venous whole blood. HbA1c is used for the monitoring of
long-term glycemic control in individuals with diabetes mellitus. For in vitro diagnostic use only.
2.6 Premier Hb9210TM Certification
In line with the ADA/EASD/IDF Working Group on HbA1c recent guidelines for reporting HbA1c, the
Premier Hb9210TM is certified to report in both IFCC and NGSP units.
• IFCC (mmol HbA1c/mol Hb)
• NGSP (% HbA1c)
For further information please contact Trinity Biotech or your local support representative.
2.7 510(K) Listing

Reference #: K112015
2.8 CE Mark
Trinity Biotech declares that the Premier Hb9210TM, related reagents and control materials conform to
the relevant provisions of the EX Council Directive 98/79/EC and Annex III, except Section 6, of the
IVDD as implemented by the European Union’s Medical Devices Regulations. The Global Medical
Device Nomenclature (GMDN) code is 30838.


PREMIER Hb9210TM
Principles of Operation
CHAPTER CONTENTS
✓ Principles of Analysis ✓ Merits of Assay ✓ Specific Performance ✓ Characteristics

Chapter 3 – Principles of Operation
3.1 Clinical Significance of HbA1c
Glycated haemoglobin (HbA1c) is of particular clinical interest in diabetes mellitus. Measurement of
glycated haemoglobin is a clinically useful means of assessing glycemic control in diabetics.6, 7 HbA1c
values reflect blood glucose levels over the circulatory half-life of the erythrocyte (about 60 days) and
correlate significantly with mean blood glucose levels during that time.4, 8 Therefore, measurement of
glycated haemoglobin provides a means, independent of multiple measurements such as patient records of
self-monitored blood glucose, for assessing the overall efficacy of therapy. Factors such as diet, exercise,
insulin regimen and stress can affect glycemic control, and therefore HbA1c values. In uncontrolled or
poorly-controlled diabetics, glycated haemoglobin values may be two or three times as high as in
non-diabetics, while meticulously controlled diabetics may have HbA1c values near or in the normal
range.9 Uncontrolled or poorly-controlled diabetics brought under better control will exhibit a gradual
drop in HbA1c values, reaching a new equilibrium in approximately eight weeks.1 There is significant
evidence that maintaining good glycemic control has a positive impact on the development of the
long-term complications of diabetes.10, 11, 12, 13, 14
3.2 Principles of Analysis

Glycated proteins (haemoglobins and plasma proteins) differ from non-glycated proteins by the
attachment of a sugar moiety(s) to the former at various binding sites by means of a ketoamine bond.
GHb and GPP thus contain 1,2-cis-diol groups not found in non-glycated proteins. These diol groups
provide the basis for separation of glycated and non-glycated components by boronate affinity
chromatography3,5. In this analytical technique, a boronate such as phenylboronic acid is bonded to the
surface of the column support. When a solution of proteins (haemolysate or diluted plasma) is passed
through the column, the glycated component is retained by the complexing of its diol groups with the
boronate (Figure 1). After the unretained non-glycated component elutes from the column, the glycated
component is eluted from the column with a reagent that displaces it from the boronate.3, 15
Figure 1: Affinity Binding of Glycated Protein
1 | Chapter 3 – Principles of Operation Rev. 23

The Premier Hb9210TM employs the principles of boronate affinity and high-performance liquid
chromatography (HPLC). The pumps transfer reagents through the analytical column which contains
aminophenylboronic acid bonded to a porous polymer support (gel).
Haemolysed samples for HbA1c analysis are automatically injected onto the column during the flow of
Premier Hb9210TM Reagent Buffer A. The glycated component binds to the boronate, while the
non-glycated component passes through the column to the spectrophotometric detector, where it is
detected at 413 + 2 nm.
After the elution of the non-glycated component, the system pumps Premier Hb9210TM Reagent Buffer
B, which displaces the glycated component from the column. The glycated component then passes
through the detector. See Figure 2 for a diagram of the binding of glycated haemoglobin in the Premier
Hb9210TM system.
Figure 2: Affinity Binding Diagram
The compositions of Elution Buffers A & B are designed to exhibit virtually identical absorption in the
413 + 2nm range to ensure a stable baseline. The detector signal is also referenced by the split-beam
technique.
In the final stage of the cycle, the column is re-equilibrated with Elution Reagent A. All reagent selection
occurs in a timed sequence designed to allow complete elution of non-glycated and glycated components.
All functions are controlled by the computer. The computer processes the signal from the
spectrophotometric detector and calculates the concentration of glycated hemoglobin or plasma protein as
a percentage of the total detected. Integration is by peak area in Absorbance Units (AU)-seconds.
The computer produces printed reports by analyzing signal as it is received by the detector. A Batch
Summary Report is printed at the end of the run.

The software, especially designed for HbA1c analysis, controls the four basic system operation functions
of sample identification, instrument operation, calculation of results, as well as printing and storing
complete reports. Based on computer hard disc capacity, many years worth of data (chromatograms and
batch summary reports) can be automatically archived.
Calculation of the percentage of GHb in the sample is by the following formula, with peak area in
AU/seconds:
(PPPPPPPP 1 AAAAPPPP PPPPPPPP + 2 AAAAPPPP
PPPPPPPP 2 AAAAPPPP)
∗ 100
The final result is obtained by comparison to reference samples using a 2-point calibration. The Premier
Hb9210TM is traceable to the IFCC method and is also NGSP certified in order to ensure A1c levels are
quantitated correctly.
3.3 Merits of the Assay

3.3.1 Boronate Affinity HbA1c Assay
HbA1c measurement by boronate affinity chromatography is free from interferences such as common
haemoglobin variants, non-glycation modifications and storage-related hemichromes.2 Boronate affinity
chromatography requires no sample pre-treatment to remove the labile (aldimine or Schiff base)
component, since only stable (ketoamine-linked) glycated haemoglobins are retained by the boronate.
3.3.2 Premier Hb9210TM
The Premier Hb9210TM combines the advantages of boronate affinity separation with the precision,
convenience and automation of high-performance liquid chromatography. After samples are loaded and
identified, the Premier Hb9210TM permits unattended operation. The Premier Hb9210TM batch runs up
to 210 samples including two dedicated positions for low and high controls, as well as the ability to run
STAT samples.
3.3.3 Glycated Haemoglobin Reporting Units
The Premier Hb9210TM system currently reports in the following units (user selectable):
✓ mmol HbA1c/mol Hb (IFCC) ✓ %HbA1c (NGSP)
Conversion Equations: mmol HbA1c/mol Hb (IFCC) ↔ %HbA1c (NGSP)
mmol HbA1c/mol Hb (IFCC) = (%HbA1c NGSP – 2.15) / 0.091516
%HbA1c NGSP = (0.0915 (mmol HbA1c/mol Hb (IFCC)) + 2.1517
Table 1: Glycated Haemoglobin Interconversions
For the latest interconversion equations between the various measurements of HbA1c reporting units,
please refer to the official NGSP website – www.ngsp.org

3.3.4 Expected Values
The expected mmol HbA1c/mol Hb or %HbA1c value for patients with diabetes will depend on physician
discretion. The American Diabetes Association (ADA)/European Association for the Study of Diabetes
(EASD) specify a treatment goal of 53 mmol/mol (7 %) or less and suggest additional action when the
HbA1c level is above 64 mmol/mol (8%).
3.3.5 HbA1c Accuracy
In accuracy testing of the Trinity Biotech Premier Hb9210TM vs. ultra2, ordinary least squares fit for the
linear regression was used with the correlation coefficient (r2) = 0.9962.
✓ The slope and intercept of the linear regression line is: y = 1.0069x - 0.1214 where the x axis are the
results
from the ultra2 and the y axis are the results from the Premier Hb9210TM.
✓ The Premier Hb9210TM demonstrates linear performance from 3.8% to 18.5% HbA1c.
Premier Hb9210 (y) vs. ultra2 (x)
19.0
c 1 A b H % , Y
17.0 15.0 13.0 11.0 9.0
Identity line A=B
y = 1.0069x - 0.1214
d o
R2 = 0.9962 h t e
7.0
M
5.0 3.0
3.0 5.0 7.0 9.0 11.0 13.0 15.0 17.0 19.0
Method X, %HbA1c
Figure 3: Method Comparison (linearity)
✓ The total number of points used in the regression was 48 per instrument (Premier Hb9210TM single
results,
ultra2 average of 2 analyses per sample).
✓ The bias calculated from the regression line at stated medical decision point of 6.5% to 8.0% HbA1c
was at
or less than 0.3% HbA1c (at generally recognized decision points).

0.6
n e e
0.5 0.4 w t e
0.3
b
s
0.2 0.1
Bias Line = -0.07
e
d o
0.0
%HbA1c c n e r e f f i D
h t e m
-0.1 -0.2 -0.3 -0.4 %
-0.5 -0.6
3.0 5.0 7.0 9.0 11.0 13.0 15.0 17.0 19.0
%HbA1c Level
Figure 4: Method Comparison (bias)
✓ The range of data included in the regression: The lowest value was 4.4% HbA1c and the highest value
was
16.2% HbA1c.
✓ The ultra2 (Method X) was the comparative method used in the regression.
✓ The standard error of estimate of the data was -0.067.
✓ The confidence interval on the overall bias is 0.047 (at the 95% CI).
✓ Ordinary least squares fit for the linear regression was used with the correlation coefficient (R2) =
0.9962.
3.3.6 HbA1c Linearity/Measurable Range
The results of this study will guide laboratory management in establishing acceptable instrument HbA1c
result reporting ranges (minimum and maximum HbA1c results allowable). As such, each lab should
verify each result is within HbA1c result linearity. The software has user-settable flags to allow each lab
to enter their proven minimum/maximum HbA1c result and values beyond this proven range are flagged
and a --- value is applied to this patient to prevent releasing a result beyond proven HbA1c linearity.
In HbA1c linearity testing of the Trinity Biotech Premier Hb9210TM, 7 samples mixed from a low and
high sample were analyzed. The Premier Hb9210TM has been demonstrated to be linear from 18
mmol/mol to 179 mmol/mol (3.8% to 18.5%) HbA1c with a correlation coefficient (r2) = 0.9958.
3.3.7 HbA1c Analytical Specificity
Several studies were performed in which normal and abnormal (>64 mmol/mol, 8% HbA1c) %HbA1c
level blood samples were directly compared to blood containing a specific potentially interfering
substance. Different concentrations of interfering substances were tested at two %HbA1c levels. As a
control method, pooled unaltered blood was analyzed. As an evaluation method, the pooled unaltered
blood was then spiked with a potentially interfering substance. The acceptance criteria for interference
was a variance in the %HbA1c value within the range of ± 5% of the original (interferant free) value. The
data is summarized below.

✓ labile HbA1c – no interference up to 1500 mg/dL
✓ icterus – no interference up to 20 mg/dL
✓ lipemia – no interference up to 3317 mg/dL
✓ carbamylated HbA1c – no interference up to 65 mg/dL
✓ acetaldehyde – no interference up to 25 mg/dL
✓ acetylsalicylic acid – no interference up to 90 mg/dL
✓ sodium heparin – no interference up to 450 USP
✓ sodium fluoride – no interference up to 1157 mg/dL (~3x industry accepted blood collection tube
concentration)
✓ EDTA – no interference up to 1440 mg/dL (~8x industry accepted blood collection tube concentration)
Hemoglobin variant (HbV) interference studies were performed using patient samples containing HbAS,
HbAD, HbAE and HbAC in an evaluation method in direct relation to the comparative method, K891235.
The acceptance criteria were that an HbV is considered to interfere if the % variance from the known
good method is greater than ± %7. No interference was observed for HbAS up to a concentration of
35.8%, HbAD up to a concentration of 28.9%, HbAE up to a concentration of 16.9% or HbAC up to a
concentration of 34.8%.
Regarding elevated levels of HbF, a 2012 publication found that HbA1c results using Trinity Biotech
Boronate Affinity showed no clinically significant interference in patients with HbF levels up to 15%18.
3.3.8 HbA1c Carryover
In linearity testing of the Trinity Biotech Premier Hb9210TM, the method has demonstrated carryover
less than 2.2 mmol/mol (0.2%) HbA1c. 20 runs were performed over 5 days of the 10 sample data set
consisting of low, decision point and high %HbA1c samples. After calculation of the cumulative means,
standard deviations, coefficients of variance, and biases of each of the pooled patient sample levels, the
following determined acceptability:
1. The coefficient of variance of each level does not exceed the acceptable product claim.
2. The standard deviation of the high-low samples should be ≤3x the standard deviation of the low-low
samples.

Cumulative Statistics Mean 5.46 %HbA1c 8.37 %HbA1c 11.23 %HbA1c SD 0.103
0.152 0.182 % CV 1.89 1.81 1.62
Lo-Lo Mean 5.41 %HbA1c Lo-Lo StDev 0.089 Hi-Lo Mean 5.51 %HbA1c Hi-Lo StDev 0.110 Pass? Yes
Table 2: Carryover Testing Data
3.3.9 HbA1c Matrix Effects – Collection Methods and Anticoagulants
Matrix comparison studies were performed to encompass the entire measuring range from 18 mmol/mol
to 179 mmol/mol (3.8 to 18.5 %) HbA1c on the Premier Hb9210TM. Different concentrations of
interfering substances were tested at two HbA1c levels. As a control method, pooled unaltered blood was
analyzed. As an evaluation method, the pooled unaltered blood was then spiked with a potentially
interfering substance. The acceptance criteria for interference was a variance in the %HbA1c value within
the range of ± 5% of the original (interferant free) value. The data are summarized below.
✓ Sodium heparin – no interference up to 450 USP
✓ Sodium fluoride – no interference up to 1157 mg/dL (~3x industry accepted blood collection tube
concentration)
✓ EDTA – no interference up to 1440 mg/dL (~8x industry accepted blood collection tube concentration)
3.3.10 HbA1c Matrix Effects – Frozen and Non-Frozen Samples
To test for matrix effects on freezing of blood sample types (venous blood samples from EDTA-treated
blood collection tubes and their corresponding hemolysate samples) for %HbA1c/glycated hemoglobin
measurements on the Premier Hb9210TM, 20 patient samples were compared as follows:
✓ Frozen hemolysate vs. non-frozen hemolysate
✓ Lysates from frozen whole blood vs. non-frozen hemolysate
✓ Frozen hemolysate vs. lysates from frozen whole blood
Analysis on the Premier Hb9210TM indicated that sample freezing had no affect on sample results. The
bias calculated from the regression line at stated medical decision point of 46 mmol/mol to 64 mmol/mol
(6.5% to 8.0%) HbA1c was at or less than 1.1 mmol/mol (0.1%) HbA1c (at generally recognized decision
points).

3.3.11 HbA1c Precision
Intra-run Precision – All sites – Haemolysate: The between-run precision study consists of three
haemolysate samples representing normal, decision point and abnormal levels of %HbA1c. They were
analyzed on 20 non- consecutive days. One internal and 2 external sites were involved, each site was
provided with the same sample set (aliquotted and frozen whole blood) and directed to perform 2 analyses
of each sample per run with 2 runs per day (a total of 80 replicates of each level per site, 240 replicates
overall. The study lasted 35 calendar days. The estimates of imprecision for each sample result obtained
from all 3 study sites (all results averaged) analysis are given in the table below:
Actual number of days involved in the experiment, and number of sites:
35 days, 3 sites
Actual total number of runs (if applicable): 120 total runs Total number of observations (including
controls): 1200 total observations
Number of instruments/devices used in the evaluation, and how results were pooled:
3 instruments, individual values not pooled
Number of reagent lots: 1 reagent lot number per reagent/column
Number of calibration cycles and calibration lots:
16 total recalibrations once study started, 1 lot of Calibrator used. Table 3: Intra-run Precision - Overview
All Labs Concentrations at which claim is made; %HbA1c 5.76
7.07 10.96 Estimate of repeatability SD, (S
r
) 0.07 0.05 0.09 Repeatability %CV = (S

r
/mean)*100 1.26 0.72 0.85 Estimate of within-device precision SD, (S

T
) 0.09 0.09 0.16 Within-device precision %CV = (S

T
/mean)*100 1.62 1.28 1.50 Table 4: Intra-run Precision – Overview


Intra-run Precision – Individual Sites – Haemolysate:
The between-run precision study data for each individual site is as follows:
Site A: External Concentrations at which claim is made; %HbA1c
5.70 7.06 10.96 Estimate of repeatability SD, (S
r

r

T

T
/mean)*100 1.84 1.58 1.62

Site B: External Concentrations at which claim is made; %HbA1c
r

r

T

T
/mean)*100 1.47 1.20 1.62

Site C: Internal Concentrations at which claim is made; %HbA1c
r

r

T

T
/mean)*100 1.55 1.04 1.27 Table 5: Intra-run Precision - Individual Sites –

Haemolysate

3.4 Chapter References
1. Bunn, H. F. et al. "The biosynthesis of human hemoglobin A1c: slow glycosylation of hemoglobin in vivo." J Clin
Invest
57:1652-1659 (1976) 2. Fluckiger, R. "Glycated Haemoglobins - Review." J Chromatog 428:279-292 (1988) 3.
Mallia, A. K. et al. "Preparation and use of a boronic acid affinity support for separation and quantitation of
glycosylated
hemoglobins." Anal Lett 14:649-661 (1981) 4. Nathan, D. M. et al. "The clinical information value of the
glycosylated hemoglobin assay." N End J Med 310:341-346
(1984) 5. Fairbands, V. F. and Zimmerman, B. R. "Measurement of glycosylated hemoglobin by affinity
chromatography," Mayo
Clin Proc 58:770-773 (1983) 6. Koenig, R. J. et al. "Correlation of glucose regulation and hemoglobin A1c in
diabetes mellitus." N Eng J Med 295:417-
420 (1976) 7. Gabbay, K. H. et al. "Glycosylated hemoglobins and long-term blood glucose control in diabetes
mellitus." J Clin End &
Metab 44:859-864 (1977) 8. Koenig, R. J. et al. "Hemoglobin A1c as an indicator of the degree of glucose
intolerance in diabetes." Diabetes
25:230-232 (1976) 9. Peterson, C. M. et al. "Feasibility of improved blood glucose control in patient with
insulin-dependent diabetes
mellitus." Diabetes Care 2:329-335 (1979) 10. Unger, R. H. "Benefits an risks of meticulous control of
diabetes." Medical Clinics of North America: Symposium on
Diabetes 66(6):1317-1325 (1982) 11. Raskin, P. R. et al. "The effect of diabetic control on the width of skeletal
muscle capillary basement membrane in
patient with Type 1 diabetes mellitus." N End J Med 309:1546-1550 12. Colwell, J. A. et al. "New concepts
about the pathogenesis of atherosclerosis in diabetes mellitus." Am J Med Glipizide
Symposium:81-84 (1983) 13. Brownlee, M. and Cerami, A. "The biochemistry of the complications of diabetes
mellitus." Ann Rev Biochem 50:385-
432 (1981) 14. Brownlee, M. et al. "Advanced glycosylation and products in tissue and the biochemical basis of
diabetic
complications." N Eng J Med 318:1315-1321 (1988) 15. Willey, D. G. et al. "Glycosylated haemoglobin and
plasma glycoprotein assays by affinity chromatography." Diabetologia
27:56-58 (1984) 16. R.R Little et al. “Interlaboratory Standardization of Measurements of Glycohemoglobin”
Clin Chem 38, 2472-2478
(1992) 17. Cas Weykamp et al. “IFCC Reference System for Measurement of Hemoglobin A1c in Human
Blood and the National
Standardization Schemes in the United States, Japan, and Sweden: A Method-Comparison Study” Clinical
Chemistry 50:1, 166-174 (2004) 18. Randie Little et al. “The effect of Increased Fetal hemoglobin on 7 common
HbA1c Assay Methods” Clinical Chemistry;
58:945-948 (2012)

APPENDIX A – EXTERNAL STUDY
Multicentre Evaluation of the Premier Hb9210TM HbA1c Analyser
W. Garry John, Randie Little, David B. Sacks, Cas Weykmap, Erna Lenters-Westra, et al., Clin Chem
DOI 10.1515, (2014)
A.1 Precision & Trueness
This multicentre evaluation was performed across five clinical laboratories located in Europe and USA;
Norfolk and Norwich University Hospital, UK (Centre 1), Diabetes Diagnostic Laboratory, University of
Missouri, USA (Centre 2), Department of Laboratory Medicine, National Institutes of Health, USA
(Centre 3), European Reference Laboratory, located in Queen Beatrix Hospital, The Netherlands (Centre
4) and European Reference Laboratory, located in Isala, The Netherlands (Centre 5).

The findings of the Multicentre Evaluation are outlined below in Table 6 - these results demonstrate the
precision of the Premier Hb9210TM boronate affinity HPLC.
CV
Mean CV for All Centres IFCC units, mmol/mol
Mean CV for all Centres NGSP Units, % Total CV (Low HbA1c) 2.71 Total CV (Low HbA1c) 1.62
Total CV (Mid HbA1c) 2.37 Total CV (Mid HbA1c) 1.59 Total CV (High HbA1c) 2.14 Total CV (High
HbA1c) 1.68
Table 6: Reported CV values reported as part of the Premier Hb9210TM Multicentre Evaluation
A.2 HbA1c Linearity/Measurable Range
The Multicentre Evaluation investigated the Premier Hb9210TM linearity at two centres. The data from
Centre 1 is shown below in Table 7 as a representative example.
SI units, mmol/mol NGSP, % units
Sample
CV
Observed Value
Theoretical Value
% Difference
Observed Value
Theoretical Value
% Difference
1 32.0 32.0 0.0 5.08 5.08 0.0 2 64.3 63.3 1.58 8.03 7.94 1.13 3 79.7 79.0 0.89 9.44 9.38 0.64 4 95.7 94.7
1.06 10.91 10.82 0.83 5 125.7 126.0 -0.24 13.65 13.68 -0.22
Table 7: Results of Multicentre Linearity Study
A.3 Interference Studies
Haemoglobin Variants
As part of the multicentre evaluation, the Premier Hb9210TM was subjected to testing in the presence of
the four most common haemoglobin variants; HbAC, AD, AE and AS. The results of this testing are
outlined below in Table 8. NOTE: NS signifies any interference observed was not significant, as noted by
the authors during the course of the multicentre evaluation.
Haemoglobin Variant
Level at which interference deemed significant
Level of interference observed (compared to secondary method) Haemoglobin AC ±7% bias 0%-0.3%
(NS) Haemoglobin AD ±7% bias -0.2% (NS) Haemoglobin AE ±7% bias -1.6%-1.7% (NS) Haemoglobin
AS ±7% bias 2.6%-2.8% (NS)
Table 8: Effects of Common Hb Variants on HbA1c Measurement on the Premier Hb9210TM

Other Potential Interferents
As well as the common variants highlighted above, further potential interferents were investigated in the
course of the multicentre evaluation. Table 9 below outlines the testing procedure utilised throughout the
course of evaluation.
Potential Interferent Level of interferent used Level of interference observed
Carbamylated Hb 0.2%-12% carbmylated Hb No significant effect
Labile HbA1c 0-120 mmol/L glucose
When glucose is 60-120mmol/L, HbA1c is 2-5 mmol/mol (0.2%-0.5% NGSP) higher. When glucose is
30mmol/L, HbA1c is ~1mmol/mol (<0.1% NGSP) high.
Low haematocrit
0.091-0.387 (Hb levels 69-140 g/L)
HCT values below 0.306 are not reliable & manual dilution is required to obtain HbA1c values Icteric
plasma Total billirubin 356 μmol/L -1.7 mmol/mol HbA1c (NS)
Table 9: Results of Interference Studies on HbA1c Measurement on the Premier Hb9210TM
A.4 HbA1c Carryover
As part of the multicentre evaluation paper the authors reported the following:
✓ “There was no effect of carryover evident from the analysis of alternate high and low samples
of HbA1c. The difference between the means of the high-low and the low-low sequence samples
was 0.6 mmol/mol (0.05% NGSP), which is less than three times the SD of the low-low values
[1.64 mmol/mol, 0.15% NGSP)].”

PREMIER Hb9210TM
Preparation for Operation
CHAPTER CONTENTS
✓ Test Components ✓ Use & Storage ✓ Preparation of Calibrators
& Controls ✓ Specimen Requirements

Chapter 4 – Preparation for Operation
4.1 Test Components
Trinity Biotech provides XL test packs in the following configurations:
XL500 Test Pack – Part Number: 09-03-0008
Component Qty/Pack Premier Hb9210TM Buffer A Reagent, 940 mL 3 Premier Hb9210TM
Buffer B Reagent, 940 mL 4 Premier Hb9210TM Diluent Reagent, 3.8L 2 Premier Hb9210TM
WASH Reagent, 940 mL 3 2 micron Pre-column Frit 1 75 micron Pre-Injection Valve Frit 1
XL500 Column 1
✓ The pack contains one 500 injection (XL500) column
✓ The pack includes sufficient reagent to perform 500 patient sample injections
✓ The configuration is designed for customers running ≤ 25,000 tests per year
Component Qty/Pack Premier Hb9210TM Buffer A Reagent, 940 mL 6 Premier Hb9210TM
Buffer B Reagent, 940 mL 8 Premier Hb9210TM Diluent Reagent, 3.8L 4 Premier Hb9210TM
WASH Reagent, 940 mL 6 2 micron Pre-column Frit 1 75 micron Pre-Injection Valve Frit 1
XL1000 Column 1 ✓ The pack contains one 1000 injection (XL1000) column
✓ The configuration is designed for customers running 25,000 – 50,000 tests per year
1 | Chapter 4 – Preparation for Operation Rev. 23
It is recommended that operators wear the appropriate PPE (personal protective equipment) while
working with potentially biohazardous materials. Furthermore, ensure to dispose of components that have
come in contact with biohazardous materials in an appropriate biohazard waste bin.

Component Qty/Pack Premier Hb9210TM Buffer A Reagent, 3.8L 3 Premier Hb9210TM Buffer
B Reagent, 3.8L 3 Premier Hb9210TM Diluent Reagent, 3.8L 9 Premier Hb9210TM WASH
Reagent, 940 mL 3 2 micron Pre-column Frit 3 75 micron Pre-Injection Valve Frit 3 XL1000
Column 3
✓ The pack contains three 1000 injection (XL1000) columns
✓ The configuration is designed for customers running ≥ 100,000 tests per year
A list of possible components associated with the system are as follows:
Reagents:
✓ Premier Hb9210TM Buffer A Reagent (Elution Reagent #1), 940 mL, (01-03-0095) ✓ Premier
Hb9210TM Buffer B Reagent (Elution Reagent #2), 940 mL, (01-03-0096) ✓ Premier Hb9210TM
Buffer A Reagent (Elution Reagent #1), 3.8 L, (01-03-0080) ✓ Premier Hb9210TM Buffer B Reagent
(Elution Reagent #2), 3.8 L, (01-03-0081) ✓ Premier Hb9210TM DIL Reagent (for Rinse Station), 3.8 L,
(01-03-0097) ✓ Premier Hb9210TM DIL Reagent (for Syringe), 3.8 L, (01-03-0097) ✓ Premier
Hb9210TM WASH Reagent, 940 mL, (01-03-0098) ✓ Purified Water
Calibrators:
✓ Hemoglobin A1c Calibrator Kit, 400μL, (01-04-0018) ✓ Hemoglobin A1c Calibrator Kit, 500μL,
(01-04-0022)
Controls:
✓ Hemoglobin A1c Control Kit, 400μL, (01-04-0015) ✓ Hemoglobin A1c Control Kit, 500μL,
(01-04-0020)
Column:
✓ Premier Hb9210TM XL1000 Boronate Affinity Column (09-06-0046)1 ✓ Premier Hb9210TM
XL500 Boronate Affinity Column (09-06-0050)1
1 Pre-Injection Valve & Pre-Column Frits (1 ea.) included with each column.

Additional items needed (not provided by Trinity Biotech):
✓ Sample vials – 13x75mm untreated test tubes for QC and haemolysate sample preparations
(03-07-0152,
Test Tubes, Vacutainer, Clear, 100/pk). ✓ Adjustable pipettor for sample dilutions – 1-200 μL range
and 1000 μL range ✓ Disposable pipette tips ✓ Purified, filtered, distilled and/or DI water for
recirculating pump head wash lines – changed weekly
Additional items recommended, but not required (not provided by Trinity Biotech):
✓ Reagent Side Trays (Left: 09-31-0010, Right: 09-31-0012) ✓ Autodiluter (capable of sampling 2 -
100μL and dispensing 200 - 1000μL) ✓ Isopropyl Alcohol
4.2 Reagent Use and Storage

1. Reagents are ready to use 2. Store at room temperature 3. Do not use beyond the expiration date printed
on the label 4. Reagents are stable for 30 days at room temperature after opening 5. Keep reagents capped
while in use 6. Reagents should be clear & colourless – do not use if cloudy or discoloured 7. Do not
pipette by mouth 8. Avoid contact with face & eyes
All unopened reagents are stable until the expiration date indicated on the container. Once opened,
Premier Hb9210TM Buffer A Reagent and/or Premier Hb9210TM Buffer B Reagent are stable for 30
days. Do not ‘top-off’ Premier Hb9210TM Buffer A Reagent or Premier Hb9210TM Buffer B Reagent.
The Premier Hb9210TM DIL Reagent and/or Premier Hb9210TM WASH Reagent are stable for 30 days.
Keep reagent bottles capped when on the instrument or in storage.
For detailed reagent installation instructions, see Section 8.2.
Note on Purified Water bottle for the Premier Hb9210TM: The function of this re-circulating liquid is that
it clears out the backside of each pump head of pump seal debris, salts etc. As such, it does not affect the
chemistry in that it never comes into contact with the other reagents or the patient sample or the route the
patient sample takes through the instrument. While the purified water will not have an impact on system
chemistry, this should be changed on a weekly basis in order to prevent re-circulation of salt debris.

4.3 Preparation of Haemoglobin HbA1c Calibrators and Controls
4.3.1 Trinity Biotech Haemoglobin HbA1c Calibrators
Directions for reconstitution of calibrators, their corresponding values and stability claims are found in
the Trinity Biotech Haemoglobin HbA1c Calibrator Package Insert.
The system needs to be recalibrated:
1. When the instrument is initially installed 2. When a column is changed 3. If control values are out of
the acceptable range stated on the package insert 4. When reference material changes values or lot
numbers
4.3.2 Trinity Biotech Haemoglobin HbA1c Controls
Controls in the normal and diabetic ranges should be included in each batch run to monitor the
performance of the system. Directions for reconstitution of controls and their corresponding target
values/ranges are found in the Trinity Biotech Haemoglobin HbA1c Control Package Insert.
4.4 Column Use and Storage

1. If a column is not to be immediately installed, it should be stored in a refrigerator at 2° - 8°C. 2. Allow
column to come to room temperature before installing on system. 3. If it is anticipated that the instrument
will not be used for a period of 7 days or more, remove the column after deactivation. Cap each end, using
the blue screw plugs provided with the column, and store at 2° - 8°C until ready for use. 4. Follow
complete instructions found in chapter 6.2 for column installation. See section 4.4.3 below for a
quick reference to column installation.
4.4.1 General Column Information
The Trinity Biotech HbA1c column is a boronate affinity column used on our HPLC analyzers. We have
high standards of Quality Assurance for acceptance and every column lot is individually tested and must
pass rigid criteria prior to shipment.
Reagents and columns are for use with the Premier Hb9210TM only. Do not substitute columns or
reagents from any other source/vendor.

4.4.2 Column Storage
The columns are shipped at room temperature in packaging that gives protection from excessively high or
low temperatures and physical damage. Upon receipt of the column in the laboratory, the columns should
be refrigerated before installation on the Premier Hb9210TM. Column refrigeration is also recommended
when the instrument is not in use for over 7 days. The columns must be protected from freezing. When a
new column is to be installed on an instrument, the old column should be removed and disposed with
biohazard waste.
4.4.3 Column Installation
When a column is changed, or a new column installed, the following quick steps will be displayed on
screen:
Figure 1: Column Change Instructions
4.4.4 Column Use
Proper use and care of the column includes software-guided start-up and shutdown of the instrument with
the column in place. These two processes clean and prepare the column for use. The start-up optimizes the
chemical balance in the column and the shutdown eliminates impurities and salt build-up in the system
and the column. Any shortcut in these processes may reduce the life of a column. Even in the busiest
laboratories, where the instrument is activated (running samples or at standby) for 24 hours per day, there
should be a regular routine to go through start-up and shutdown once per day.
Attach the tubing in a manner so that the end of the tubing is sealed tightly to the column fitting. It is
imperative that there are no void volumes between the end of the tubing and the column inlet or outlet.

4.4.5 Summary
Keeping the instrument well maintained and practicing routine daily maintenance will ensure that the
expected life of the column is met. Laboratory protocols should ensure columns are not changed unless
the full life has been achieved. As always, please contact Trinity Biotech or your local support
representative should you have any questions.
4.5 Specimen Requirements

4.5.1 Collection
Whole blood samples should be aseptically collected by venipuncture or capillary collection. EDTA is the
preferred anticoagulant. While Heparin and fluoride are also acceptable, the use of other anticoagulants
on the Premier Hb9210TM is not recommended – for further information, refer to Chapter 3 of this
manual.
4.5.2 Patient Sample Storage Stability
In a 360 day study performed for the Premier Hb9210TM’s FDA 510k filing, a sample stability study
with Whole blood (and associated haemolysate) specimens at 3 different HbA1c levels were analyzed by
the Premier Hb9210TM boronate affinity assay method on Day 0, then divided into aliquots stored at five
different temperatures (37°C, 20-25°C, 2-8°C, -20°C and -70°C) for analyses on subsequent sampling
intervals. Acceptance limits are considered acceptable if results fall within 5% bias from time zero
%HbA1c result. Any sample falling outside 5% mathematical bias from time zero is considered
unacceptable.
4.5.3 Premier Hb9210TM Sample Storage Stability Study
Whole blood and haemolysate storage stability claims are listed in the table below:
Sample Storage Temperature Sample type -70°C -20°C 2-8°C
20-25°C 37°C Whole blood 360 Days 30 Days 30 Days 6 days 1 day Hemolysate 15 Days 10 Days 30
Days 6 days 1 day
Figure 2: Sample Storage Temperature Claims

Stability Data
Figure 3: Haemolysate Storage Stability
Figure 4: Whole Blood Storage Stability
4.5.4 Anaemic, Normal, pRBC and Washed RBC Sample Dilutions
When compared to a normal haematocrit blood sample, on-line (undiluted blood samples) and off-line
(haemolysate samples) dilutions of samples with low haemoglobin concentration (‘anaemic’ blood
samples) at HbA1c levels of 29mmol/mol, 64 mmol/mol and 80mmol/mol (4.85%, 8.0% and 9.5%) can
be considered equivalent when assayed on the Premier Hb9210TM analyzer, the bias on results from the
same sample at normal haemoglobin concentration was less than 5%.
(haemolysate samples) dilutions of samples with high haemoglobin concentration (pRBC blood samples)
at %HbA1c levels of 29mmol/mol, 64 mmol/mol and 80mmol/mol (4.85%, 8.0% and 9.5%) can be
considered equivalent when assayed on the Premier Hb9210TM analyzer, the bias on results from the
same sample at normal haemoglobin concentration was less than 5%.
(haemolysate samples) dilutions of samples with high haemoglobin concentration that were washed with
saline (washed pRBC blood samples) at %HbA1c levels of 29mmol/mol, 64 mmol/mol and 80mmol/mol
(4.85%, 8.0% and 9.5%) can be considered equivalent when assayed on the Premier Hb9210TM analyzer,
the bias on results from the same sample at normal haemoglobin concentration was < 5%.

4.5.5 Sample Racks
The Premier Hb9210TM sample racks come in 3 bright colours so that they are easily distinguishable
from one another.
Figure 5: Premier Hb9210TM Sample Racks
✓ White – Whole Blood Patient Samples ✓ Pink – Haemolysate Samples ✓ Yellow – Anaemic
Samples
To measure HbA1c, either mixed whole blood or packed red blood cells (pRBC's) may be used for
on-line (instrument prepared) or off-line (hand pipette) dilutions. Packed red blood cells (pRBC’s) are
defined as follows:
✓ Blood samples settled naturally by gravity, not by centrifuge ✓ There is a uniformly clear plasma
layer (golden colour) ✓ The pRBC layer is not brown in colour ✓ There is no evidence of lysed RBC’s
(red colour in the plasma layer) ✓ The sample has a normal haematocrit (approximately half the sample
height in the settled blood tube is
pRBC (packed red blood cells), half the sample height is clear plasma ✓ The sample has no obvious
clots, lipemia or other noticeable abnormalities (extremely lipemic samples can
be washed 5 times with saline)
Minimum Sample Volume: A minimum of 0.5mL haemolysate, 0.5mL packed red blood cells or 1mL
mixed whole blood is typically required for analysis.

The appropriate dilutions to be used with the Premier Hb9210TM are as follows:
Whole Blood Sample Rack Function:
✓ The Premier Hb9210TM will automatically dilute settled whole blood samples (including anaemic
patient samples with a sufficient-to-sample pRBC layer) to a 1:150 dilution when this particular function
is chosen on the instrument.
Haemolysate Sample Rack Dilution Requirements:
Recommended haemolysate dilutions are as follows:
✓ Anaemic Sample Haemolysate Preparation: 1:75 manual (off-line) dilution of a mixed whole blood
tube of an
anaemic sample ✓ Normal Sample Haemolysate Preparation: 1:150 manual (off-line) dilution of a
mixed whole blood tube of an
normal haematocrit sample ✓ pRBC Haemolysate Preparation: 1:300 manual (off-line) dilution of a
non-mixed whole blood tube of an
anaemic sample
Anaemic Sample Rack Function:
Anaemic samples feature lower red blood cell counts than normal haematocrit patients, as such the
Premier Hb9210TM anaemic rack automatically dilutes non-settled anaemic patient blood samples to a
1:75 dilution in order to facilitate a more concentrated sample injection. While most times, an anaemic
sample can be analyzed in the whole blood rack if a sufficient-to-sample pRBC layer exists, there may not
be enough pRBC’s to sample (and the total area of the sample may not be above the minimum acceptable
total area). In this case, the tube can be mixed, placed in the Anaemic rack, then this rack immediately
analyzed. The anaemic rack dilution parameter increases the overall amount of sample taken for analysis.
If mixing the anaemic sample prior to placing in the anaemic rack yields a total area below the minimum,
an off-line dilution of the sample is required. The final dilution can be manually adjusted to yield a total
area within acceptable range of the instrument.

4.5.6 Common Errors with Sample Tubes
Common sample loading and sample type-related errors (and how to avoid them) on the Premier
Hb9210TM are as follows:
1. Whenever possible, place desired sample tube barcode sticker towards the left opening of the rack.
2. Do not place whole blood samples in the H (haemolysate) rack. This will inject a non-diluted sample
into the system. The samples total area will be above maximum acceptance limits. Perform multiple
baseline calibrations to wash system out and ensure a flat baseline.
3. Do not place haemolysate samples in the W (whole blood) or A (anaemic) racks. Placing diluted
samples (haemolysates) in a W (whole blood) or A (anaemic) rack will result in a sample that is too
dilute. Ensure to utilize correct rack for each sample type.
4. Do not place (or allow) more than 3 barcode stickers on a sample tube. Too many stickers on sample
tube can lead to spatial issues when attempting to place in a sample rack. As outlined in point 1, the
barcode sticker should be pre-aligned to the left so no tube rotation is required to read barcode sticker.
5. Remove test tube septa (‘caps’) on the following sample types:
a. Calibrator and Control samples b. Haemolysate samples that will be repeatedly sampled by the system
4.6 Limitations of the Assay

In addition to the interferents listed in Chapter 3, the following are limitations of the HbA1c assay:
✓ Abnormal RBC Survival - Correct HbA1c analysis depends on stable haemoglobin with the typical
lifespan. As with all HbA1c analytical methodologies, patients with shortened RBC life, such as occurs in
haemolytic anaemias, will exhibit decreased HbA1c values, with the decreased life span dependent upon
the severity of the anaemia and/or the presence of unstable haemoglobins like Hb SS, Hb CC and Hb SC.
Patients with extended red cell life, such as occurs in polycythemia or after splenectomy, may exhibit
increased HbA1c values. Iron deficiency anaemia may yield falsely high results. Glycated Plasma Protein
(GPP) and/or fructosamine assays should be considered under such conditions.
✓ Degraded Samples – Do not freeze/thaw samples more than once. Do not keep samples beyond the
sample stability data shown in section 4.5.3 above. Because the potential exists for thawing in a
“frost-free” freezer, when possible, avoid storage of specimens in a frost-free freezer. Frozen samples (-20
to -80° C) should be examined for clarity. Haemolysates that appear cloudy or exhibit precipitation should
be filtered (.45 micron) or centrifuged prior to analysis. This protects the instrument lines and extends the
life of the frit and analytical column. If longer-term storage is required, each laboratory should investigate
sample stability in its own facility.


PREMIER Hb9210TM
Hardware Components
CHAPTER CONTENTS
✓ Introduction ✓ Module Information ✓ Decontamination Protocol
for Service & End of Life

Chapter 5 – Hardware Components
5.1 Introduction
This section defines environmental, physical, electrical requirements, system description and basic
operation of the Premier Hb9210TM System, which is comprised of the Autosampler, Pump, Detector,
and CPU for data handling system. All components of the Premier Hb9210TM are controlled through the
Premier Hb9210TM Application Software. System electrical requirements can be found in Chapter 1, Site
Preparation.
5.1.1 A Note on Using Manufacturer-Approved Replacement Parts
Instruments must be serviced in accordance with the manufacturer’s service schedule and with only
manufacturer approved and supplied replacement and maintenance parts – these are described further in
this chapter. Any changes that are made, or applied, outside of the original manufacturer’s guidance or
service schedule represents off-label use of the product and voids all warranties, voids the product
registration and clearance status and becomes the legal responsibility of the site and persons making the
unauthorized alterations to the medical device product.
Figure 1: Premier Hb9210TM System
1 | Chapter 5 – Hardware Components Rev. 23

5.2 Autosampler System
The Autosampler System consists of a Sample Transport Module, the Probe Module and Injection Valve
Module.
5.2.1 Sample Transport Module Description (09-30-1300S)
The Sample Transport Module is designed to hold a maximum of 21 racks (each linear rack holds 10
sample tubes). There are 3 sample rack types: Whole blood, haemolysate and anaemic. Each is brightly
coloured, has a unique identification letter and number with corresponding barcode.
• W is for whole blood, these racks are white with a Red identification tab.
• H is for haemolysate (1:150 off-line dilution), these racks are coloured pink.
• A is for anaemic (a more concentrated haemoglobin final dilution will be prepared automatically) and
these racks are coloured yellow.
Figure 2 below shows the transport module with the front cover removed
Figure 2: Premier Hb9210TM Sample Transport Module
All racks hold standard 13x75mm sample tubes with septum (BDTM and SarstedtTM standard). Whole
blood samples do not need to be pre-mixed. The QC materials are held in a 5-position QC wheel for fully
customizable within-run, repeated QC sampling of a single haemolysate preparation of each material
Do not utilize any sample racks other than those provided by Trinity Biotech. Failure to do so may cause
sampling errors, sample identification errors, probe errors/damage and/or sample transport errors/damage.

5.2.2 Probe Module Description (09-30-1620S)
The Probe Module (shown in Figure 3) consists of a septum piercing probe and an integrated sample
preparation station (‘diving board’). The probe is a septum piercing probe operating in the z-axis. Under
the probe, the sample preparation station moves in and out, allowing the probe to enter 4 distinct positions
(starting from the back):
1. Active Rinse Station (ARS) – Sample dilutions and probe rinsing occurs here. An integrated Premier
Hb9210TM DIL reagent pump flushes the ARS between samples. 2. Injection Port – Once the diluted
sample is obtained (either whole blood diluted in the rinse station or a manually prepared haemolysate
sample), it is injected into the injection port which is connected to the sample loop. After the injection, the
injection port tubing and the injection port top are rinsed out with Premier Hb9210TM DIL reagent in
preparation for the next sample. 3. Probe guide – Steadies the probe while it samples blood tubes in the
sample transport area and also aids in probe withdrawal from sample septum. 4. STAT Sample – Whole
blood, haemolysate, anaemic whole blood or Sarstedt 13mm tubes, as chosen through the software
interface
Figure 3: Premier Hb9210TM Probe Module
Do not remove the Probe Module from analyzer by pulling on the probe needle column. Utilize the
handles on either side of the probe. Failure to do so may cause sampling errors.

5.2.3 Injection Module Description (09-30-1900S)
The Injection Module consists of a 2-position 6- port injection valve with 5-μl sample loop positioned
vertically (between positions #1 & #4). The sample loop allows the diluted blood sample to be introduced
into a fixed volume sample loop at low pressure. Upon switching the valve, the pump fluid stream is
directed through the loop to push the sample into the column and detector. After several seconds, the
sample has been injected and the valve switches back in order for the sample loop to be cleaned, and the
next sample loaded. The sample is introduced from the upper left side (position #2) and is pre- filtered
before entering the injection valve by a 75μm frit. The injection sample waste is expelled from the lower
left side (position #3), the reagents from the pump are introduced from the lower right side (position #5)
and the sample frit and column pre-heater is connected at the upper right side (position #6).
Figure 4: Injection Module Position Map
5.3 Pump Module (09-30-2000S)

5.3.1 System Description
The pump is a microprocessor controlled binary system capable of delivering gradient reagent profiles.
On analyzers with serial numbers prior to 200734, the system uses a Reagent Manifold to automatically
switch to Wash reagent to flush the pumps, tubing, column, and flowcell when the system performs an
automated shutdown. On analyzers with a serial number of 200734 or later, the gradient is delivered
automatically without a Reagent Manifold.
A wash-channel on the pump head helps keep the pistons clean by flushing them continually as the
system is pumping. A static mixing tee is utilized to provide mixing of the reagents. A Syringe Prime
Valve is installed on the Pump Module in order to prime the system during the activation cycle.
Figure 5: Premier Hb9210TM Pump Module

5.4 Oven Module (09-30-1800S)
5.4.1 System Description
The Oven Module is located above the Injection Valve. The column heater ensures steady boronate
affinity column temperature. At the left of the oven is a cylindrical heat exchanger containing ≈ 0.5 meters
of 0.010”I.D. tubing to pre-heat the incoming reagent. The oven module is shown in Figure 6.
5.4.2 Performance Specifications
✓ Temperature Range: 30-70°C ✓ Temperature Stability: ±0.2°C ✓ Temperature Repeatability: ±0.5°C
✓ Temperature Accuracy: ±0.5°C ✓ Safety Cutout: 70°C
Figure 6: Premier Hb9210TM Oven Module, Closed (left) & Open (right)
5.5 Detector Module (09-30-1700S)

5.5.1 Detector Description
The detector is an LED-based microprocessor controlled wavelength absorbance detector. The LED lamp
is set at the appropriate wavelength for the Premier Hb9210TM System operation (413 ±2nm). The
detector has sensitivity range of 0.001 to 2.0 AUFS (Absorbance Units, Full Scale). The detector is
controlled via the Premier Hb9210TM Application Software.
✓ For detector maintenance steps, refer to
section 8.4 of this manual.
Figure 7: Premier Hb9210TM Detector Module

5.6 CPU Module (09-30-1500S)
5.6.1 Data Handling Description
The data handling system consists of a CPU, colour touch screen monitor, printer, and a handheld barcode
gun combined with the Premier Hb9210TM Application Software. The CPU is running with a Windows 7
or XPTM-embedded operating system. A complete description of the operation of the Software can be
found in Chapter 6 of this manual.
5.6.2 Physical Characteristics Due to the rapidly changing nature of today’s computer, specific parameters
will not be included in this manual. If you require information on your specific computer, please contact
Trinity Biotech or your local support team.
✓ For steps on aligning the touch screen monitor responsiveness, please refer to sections the
Troubleshooting section of this manual.
Figure 8: Premier Hb9210TM Data Handling System (with covers removed to show CPU module)
Do not utilize software that is not supplied by Trinity Biotech on your Premier Hb9210TM system. CPU
resource issues and/or malicious software routines (‘viruses’) can introduce errors in system operation. If
there are any questions, contact Trinity Biotech or your local support representative.

5.7 Barcode Readers
5.7.1 Manual Barcode Reader Gun
The barcode reader gun is used for scanning in of barcoded consumables. This information can also be
manually inputted if required – for further details please refer to the Software section of this manual.
The barcode reader gun can be installed by simply connecting the USB to the 4-port USB hub on the
Premier Hb9210TM Also supplied is an optional stand for the barcode gun.
Part Numbers:
Barcode Scanner – 09-31-0011
Stand for Barcode Reader – 09-31-0013
Figure 9: Barcode Gun

5.7.2 Automatic Barcode Reader Assembly (on Sample Transport)
The barcode reader on the sample transport identifies the sample rack type (H – Haemolysate, W – Whole
Blood or A – Anaemic) and also identifies the sample ID from the barcode label on the sample tube.
Figure 10 shows the correct (and incorrect) alignment of the barcode reader on the sample barcode label
(example A is the only correct read).
Contact your Technical Service Representative to optimize barcode and rack alignment if sample
barcodes are not being correctly identified. When the barcode reader is properly aligned and the test tube
barcode sticker is visible on the left side of the rack, the read rate is typically >95%.
Figure 10: Examples of Correct & Incorrect Barcode Reader Alignment
5.7.3 Barcode Sticker Application
Barcode stickers should be affixed to the sample tube vertically. Diagonal placement or a barcode label
with wrinkles could result in reading errors. It is also important that proper printing specifications be
followed for barcode labels. If you have further questions about printing resolution for your barcode
labels, contact your printer manufacturer.
The barcode label should be placed more than 20mm from the bottom of the tube and with about 5mm of
space above the tube. Barcodes should not exceed 75mm in length. Dimensions of proper barcode sticker
placement are shown in Figure 11.
Figure 11: Barcode Label Placement Dimensions

5.8 Automatic Debubblers & Waste Pump

Instruments from serial number 200734 onwards will have the following hardware additions:
5.8.1 Automatic Debubblers
The Auto-debubblers eliminate the need for manual debubbling as part of periodic operator maintenace.
The manual debubblers are no longer present directly under each Pump Head inlet.
The Auto-debubbling function occurs automatically after the system is primed either during system
activation or manually through the analyser software. The new Auto-debubblers are located under the
Right Side Panel, as shown in Figure 12.
Figure 12: Auto-debubbler Hardware

5.8.2 Waste Pump
The automatic Waste Pump hardware eliminates the need for a gravity-driven waste system. The Waste
Pump automatically triggers on every time the Active Rinse Station has been activated. The new
automatic Waste Pump hardware is located under the Rear Lower Panel, as shown in Figure 13.
Figure 13: Waste Pump Hardware

5.9 Decontamination Protocol for Service and End of Life Disposal
The Premier Hb9210TM system should be decontaminated prior to service, shipping or end of life
disposal. Please complete the decontamination prior to servicing. Unless specified, the Service Engineer
will still need to follow all biohazard precautions, as usual.
The decontamination process is as follows:
1. Prior to cleaning, utilize the appropriate PPE (Personal Protective Equipment), such as gloves, face
shield/goggles and a lab coat. 2. Clean probe and injection port area with 70% (or higher) alcohol
swab. Dispose swab in biohazard collection
container. 3. Wipe down external surfaces with cleaning solution (a 10% solution of household bleach
in warm tap water – made less than 7 days prior) and allow to air dry. When cleaning, paying close
attention to high-touch areas, including:
a. Touch screen b. Keyboard c. Sample transport area
d. External covers e. Barcode scanner f. Sample racks
4. Perform a manual enzyme cleaning routine (if applicable), then send system through a standard
shutdown/deactivation.
For End of Life Disposal, return module(s) and/or instrument to your distributor, according to WEEE
directive 2012/19/EU, if applicable. The Waste Electrical and Electronic Equipment Directive (WEEE
Directive) is the European Community directive 2012/19/EU on waste electrical and electronic equipment
(WEEE) which, together with the RoHS Directive 2012/19/EU, became European Law in August 2012,
setting collection, recycling and recovery targets for all types of electrical goods.
The important guidelines for the WEEE directive are as follows:
1. The requirement not to dispose of WEEE as unsorted municipal waste and to collect such WEEE
separately;
2. That return and collection systems are available
by your Trinity Biotech distributor;
3. That each laboratory has a role in contributing to reuse, recycling and other forms of recovery of
WEEE;
4. That there are potential effects on the environment and human health as a result of the presence of
potentially hazardous substances in electrical and electronic equipment;
5. That the symbol shown below means to not dispose of any parts of this item in unsorted municipal
waste and that this device contains WEEE that should be specially collected.
Figure 14: WEEE Directive Symbol

PREMIER Hb9210TM
Software & Operation
CHAPTER CONTENTS
✓ Start Up ✓ Preparation for
Patient Sample Runs ✓ Menus & Software
Tools ✓ Data Directories

Chapter 6 – Software & Operation
6.1 Introduction & Start Up
The software for the Premier Hb9210TM is controlled through a Microsoft Windows 7 or XPTM
embedded program especially designed for Trinity Biotech HbA1c analysis. The software is installed on
the hard drive of the computer system and all analysis information is also stored there.
6.1.1 Definitions
Click – On the computer screen, go to (or on top of) the subject with the (on-screen mouse pointer) and
click once with the left mouse button or tap once on the touch screen with your finger.
Double-Click – Go to the subject with the (on-screen mouse pointer) and quickly click twice with the left
mouse button or quickly tap twice on the touch screen with your finger.
Right-Click – On the computer screen, go to (or hover over) the subject with the (on-screen mouse
pointer) and click once with the right mouse button. There is no touch-screen function for right click.
After the Premier Hb9210TM is initially installed; a Trinity Biotech or local support technical service
representative will assist with the initial start-up. All of the following ‘edit’ screens are readily available
through the software if any variables need to be modified after installation. They will also be revisited
whenever software updates are installed.
Once the instrument has been powered up, and software installed, the Trinity Biotech or local technical
service representative will open the Premier icon located on the desktop (shown in Figure 1).
Figure 1: Premier Hb9210TM desktop icon
1 | Chapter 6 – Software & Operation Rev. 23
Do not utilize software that is not supplied by Trinity Biotech on your Premier Hb9210TM system. CPU
resource issues and/or malicious software routines (‘viruses’) can introduce errors in system operation. If
there are any questions, contact your Trinity Biotech representative.

The password screen will appear once the program is launched, and will continue to appear any time the
program is opened, or following deactivation. Enter your password and continue to the installation
screens.
✓ Enter your password. If you have not assigned a password yet, the default password is 7235. ✓ Touch
or click the green checkmark on
dialogue below to continue:
Figure 2: Login Prompt
6.2 Installation Screens

When the Premier Hb9210TM software launches for the first time, a number of dialogue boxes will allow
you to edit system variables before analysis of patient samples will be able to occur.
✓ Touch or click the green checkmark on
dialogue below to continue:
Figure 3: New Software Installed Dialogue

✓ Enter the desired Run Parameters in the Run Parameters dialogue box (shown in Figure 4).
✓ Touch or click the green checkmark to
continue.
Figure 4: Run Parameters Dialogue
✓ Enter desired System Parameters in the System Parameters dialogue box (shown in Figure 5).
✓ Touch or click the green
checkmark to continue.
Figure 5: System Parameters Dialogue

✓ In the Lot Numbers dialogue box, using the barcode gun, or by means of manual type- entry, enter
current lot number information.
✓ Each Premier Hb9210TM HbA1c analytical column, Trinity Biotech package insert for Calibrators
and Controls and each Premier Hb9210TM reagent bottle have a unique barcode to facilitate quick and
easy input of set points and lot number information.
Figure 6: Column Lot Number Entry Dialogue

✓ When column barcode is scanned, the quick steps in Figure 7 will be displayed on screen:
Figure 7: Column Change Instructions
✓ Across the top are various tabs for each consumable item with a lot number. Once done entering lot
numbers press the green check mark to save and exit.
Figure 8: Calibrator Lot Number Entry Dialogue

✓ To use the barcode gun, select the barcode icon on- screen and proceed to scan the required barcode.
For customers using third-party HbA1c calibrators and/or controls, values can be entered manually for the
user-determined set points and range.
Figure 9: Control Lot Number Entry Dialogue
✓ The dialog boxes for the lot number screens are shown in Figures 10 to 14.
Figure 10: Buffers A & B Lot Number Entry Dialogue

✓ Upon successful barcode entry of the first reagent, the system will display the following dialog box
Figure 11: Reagent Bottle Change Instructions
Figure 12: Diluent Lot Number Entry Dialogue

Figure 13: Wash Lot Number Entry Dialogue
✓ Lot Numbers Log: A historical list of lot numbers are kept in the Lot Numbers Log.
Figure 14: Lot number Entry Log

✓ In the Control Scheme dialogue box, choose desired method of running in-run Control Material
✓ Touch or click the green
checkmark to continue.
Figure 15: Control Scheme Dialogue
✓ The main Premier Hb9210TM assay screen will appear as soon as all system variables have been
entered. Click
on the Main Menu (User Interface) icon on the toolbar; upper left icon highlighted in Figure 16 below.
Figure 16: Main Assay Screen

✓ From the Main Menu screen,
touch or click the ‘Edit’ button.
Figure 17: Main Menu Selection
✓ From the Edit Menu screen, touch or click on the Peak Analysis & Results Codes icon.
Figure 18: Edit Menu - Peak Analysis & Results Codes Selection

✓ The Peak Analysis & Results Codes dialogues are designed to assist the operator in alerting him/her to
potential issues with both patient sample runs as well as calibration runs. While a number of Peak
Analysis & Results Codes values are user-definable, a number cannot be amended from the default
settings and accordingly will not be amendable. There are two PARC dialogues, Calibration Run and
Samples Run, and these can be used to define parameters for the appropriate analytical runs.
✓ Default values, appropriate for software versions, will be loaded. The icon at the lower left of the menu
(highlighted in Figure 19) can be used to reset defaults if values are incorrectly changed.
Figure 19: Reset to Defaults Icon
✓ Touch or click the green checkmark to continue.
Figure 20: Peak Analysis & Results Codes Dialogue (Patient Samples)

Figure 21: Peak Analysis & Results Codes Dialogue (Calibration Run)
The next sections will aide you through an initial Background & System Calibration. Upon completion of
the Baseline and HbA1c Calibrations, the Premier Hb9210TM will be ready to run analysis of patient
samples.

6.3 Preparation for Patient Sample Runs
6.3.1 Activate System
✓ Return to the Main Menu screen. To do this, touch or click the ‘Return to Main Menu’ icon in the
bottom right of the Edit Menu (shown in Figure 22).
Figure 22: Return to Main Menu Icon
✓ From the Main Menu screen, touch or click on the Operation icon.
Figure 23: Main Menu - Operation Selection
The Trinity Biotech Premier Hb9210TM features an automatic sampling and injection system that allows
full walk-away capability following the commencement of an analysis run.

✓ From the Operation Menu screen, touch or click the Activate System icon.
Figure 24: Operation Menu - Activate System Selection
✓ If the message in Figure 25 appears, scan in appropriate lot numbers. To continue, select the green
checkmark.
Figure 25: Scan Lot Numbers Dialogue
✓ If from the Run Parameters window a Calibration Reminder has been activated, the system will
prompt the operator to run a calibration.
Figure 26: Calibration Reminder Dialogue

6.3.2 System Calibration
✓ Upon completion of the System Activation, the Premier Hb9210TM will be in Standby mode. ✓
Return to the Main Menu by touching or clicking the ‘Show User Interface’ button on the top left of the
main
Premier Hb9210TM assay screen. ✓ From the Main Menu screen, select the ‘Operation’ icon. ✓
From the Operation Menu, select the ‘Run Calibration’ option shown below. This process includes a
baseline noise calibration to cancel out any system noise (e.g. Pumps, other vibrations) from the final
chromatography.
Figure 27: Operation Menu - Run Calibration Selection
✓ The Calibration Run dialogue shown in Figure 28. NOTE: The 1x option has been disabled. Only the
2x and
3x options are available (for improved calibration accuracy).
Calibration of the Premier Hb9210TM is fully automated and calibration can be initiated with the touch of
a button. Once a calibration run has been initiated Calibrators loaded on the QC wheel, as well as
Baseline noise calibrations, are used to establish appropriate analytical conditions for optimal sample
results.

✓ By selecting the Run Controls Only option, the system will simply run Control I, then Control II (and
not the calibrators).
✓ For information on the Peak Analysis & Result Codes that ensure improper calibration is alerted to the
operator, please refer to section 9.3.2.
Figure 28: Run Calibration Confirmation Dialogue

6.3.3 Baseline Calibration
✓ Although baseline noise cancelation is included in each Calibration run, Operators also have the option
to run an independent Baseline Calibration should their local protocol require it, or as part of any
troubleshooting or maintenance activities. ✓ Return to Main Menu by touching / clicking ‘Show User
Interface’ button on top left of the main Premier Hb9210TM assay screen. ✓ From Main Menu screen,
select the ‘Operation’ icon. ✓ From Operation Menu, select the ‘Run Baseline’ option shown in Figure
29.
Figure 29: Operation Menu - Run Baseline Selection
The Baseline Run dialogue is shown in Figure 30.
Figure 30: Run Baseline Confirmation Dialogue

Once the Run Baseline button has been selected, the Premier will perform 3 Re-equilibration injections
and 2 Baseline calibration injections.
See Figure 31 for an example of what a baseline injection will look like on-screen.
Figure 31: On-screen Baseline Report
Do not be alarmed if the baseline is not completely flat. The baseline movement is caused by system noise (eg.
Pumps, other vibrations) and is picked up by the detector. The purpose of running a baseline is to calibrate this noise
out of the final chromatography.

6.3.4 Load Racks
Figure 32: Load Racks Icon
✓ If running more than 11 racks of samples, the Load Racks icon highlighted in Figure 32 can be
selected. This will allow racks loaded in the front to be transported to the back to facilitate further loading
of racks.
Figure 33: Load Racks Dialogue
6.3.5 Patient Sample Run
To start an automated Patient Sample run:
✓ From the Main Menu screen, touch or click the ‘Operation’ icon.
✓ From the Operation Menu, select the ‘Run Samples’ option shown in Figure 34.
Figure 34: Operation Menu - Run Samples Selection
When running samples without barcodes (identified as ‘noID’ samples in software), the end of run (EOR)
tube needs to be inserted after the last sample. If the operator wants to re-use the same racks during a
batch run, no EOR tube is needed. The racks can be refilled with new samples, which will then be
injected onto the column and the instrument will continue to record data.

✓ Prior to commencing a patient sample run, successful baseline calibration and system calibration must
be carried out. Further to this, QC material must be analysed and demonstrated to be within the package
insert values inputted into the system.
✓ When Control Values are acceptable, patient whole blood samples are now ready to be analyzed. The
Run Samples dialogue is shown in Figure 35.
✓ Select an End of Run Status (either Stand by or Shutdown) by selecting either of the radio buttons.
✓ NOTE: the Est. # of injections to run field informs the operator as to how many injections can be run
based on the current estimated reagent levels, i.e. how many can be run before a reagent change is
required.
Figure 35: Run Samples Confirmation Dialogue
Figure 36: Example of Final Chromatogram from the Main Assay Screen
✓ The last chromatogram of the Sample Run is shown on screen, as shown in Figure 36.

6.3.6 Automated Operator Assistant
The Automated Operator Assistant, in the top right corner of the Run Samples dialogue – is designed to
improve laboratory efficiency. The Premier Hb9210TM operator is able to begin their daily routine with
only navigating to one application software menu – Activation, running a System Calibration (which
includes Baseline runs) & beginning a patient sample run can commence with the touch of a button.
When initiating a system run, the Operator can automatically select System Activation, System
Calibration & Begin Sample Run from the options to right of the ‘Run Samples’ dialogue as shown in
Figure 37.
✓ For information on the Peak Analysis & Result Codes that ensure improper sample chromatography is
alerted to the operator, please refer to the Troubleshooting section of this manual.
Figure 37: Automated Operator Assistant Dialogue

6.4 Deactivate System/Exit Application
The system must be INACTIVE to exit application. The end of run status can be set at the beginning of
each run. To deactivate the system from the STANDBY condition:
✓ From the Main Menu screen, touch or click the ‘Operation’ icon.
✓ From the Operation Menu, select the ‘Deactivate System’ option shown below.
Figure 38: Operation Menu - Deactivate System Selection
Figure 39: Auto-Deactivation Specifications
✓ The Premier Hb9210TM can also be automatically deactivated at a specific time of day or after a
specified amount of time in Standby Mode. This can be customized in the Run Parameters dialogue.

✓ Once the system is INACTIVE, to exit the Premier Hb9210TM application, from the Main Menu,
select the ‘Exit Application’ button.
Figure 40: Operation Menu - Exit Application Selection
✓ Click the green checkmark to
exit the application.
Figure 41: Exit Application Dialogue
This concludes the Introduction & Start-Up routine in order to be able to begin to run patient samples.
The next section will outline the remaining sections of the Graphic User Interface.

6.5 Other Menus & Software Tools

6.5.1 Operation Menu
The Operation Menu & all of its corresponding options can be found below.
✓ Activate System – Activates the Premier Hb9210TM Analyzer to prepare for patient sample analysis.
Effectively equilibrates the column to reach optimal conditions for chemistry to occur. Current activation
time is 25 minutes.
✓ Deactivate System – Deactivates the Premier Hb9210TM Analyzer. Pumps Wash Reagent throughout
the system in order to remove any salt residues and to prepare the column for overnight storage. Current
deactivation time is 20 minutes.
Figure 42: Operation Menu
✓ Emergency Stop – An ‘Emergency Stop’ will send the Premier to INACTIVE mode. In order to
perform an Emergency Stop, touch or click the red octagonal button shown below in Figure 44. This
button is only used as emergency stop if the Premier is at Standby. If the Premier is running samples, the
button is primarily used as an ‘Abort Run’ function – the ‘Abort Run’ function will send the system to
Standby mode, from which an ‘Emergency Stop’ can then be performed. Emergency Stop is also
available from the Operation Menu (see Figure 43).
Figure 43: Emergency Stop Button – Operation Menu

Figure 44: Emergency Stop Button - Main Assay Screen Selection
✓ Click OK on dialogue below to have System
Perform an Emergency Stop.
Figure 45: Emergency Stop Confirmation Dialogue
✓ Run Samples – See Section 6.3.5
✓ Run STAT Samples – The STAT sample position is located on the end of the probe module’s moving
arm (also containing the injection port and sample mixing station)
Within a run, the user is able to run a STAT sample. When this is selected, analysis of the current sample
will be completed before the STAT sample is immediately run. Only one patient name can be entered per
STAT sample analysis. To do so, the user needs to select the STAT Sample icon from the main software
screen, or from the Operation Menu. It is notated by an ambulance, as shown in Figure 46. Once selected,
the following screen will appear:
Figure 46: STAT Sample Icon

Figure 47: STAT Sample Dialogue
✓ Abort Run - In order to abort a patient sample run, touch or click the red octagonal button shown
below. The Premier will finish the current sample being analyzed and then go to Standby. Selecting the
red octagonal button twice will result in an ‘Emergency Stop’ (see ‘Emergency Stop’ above).
✓ There is also an Abort Run option in the
Operation menu, as shown in Figure 48.
Figure 48: Abort Run Icon - Operation Menu
Figure 49: Abort Run Button - Main Assay Screen Selection

Figure 50: Abort Run Confirmation Dialogue
✓ Run Calibration – See Section 6.3.2
✓ Run Baseline – See Section 6.3.3
6.5.2 Edit Menu
The Edit Menu & all of its corresponding options can be found below.
✓ Run Parameters – See Section 6.2
✓ System Parameters – See Section 6.2
✓ Lot Numbers – See Section 6.2
✓ Control Scheme – See Section 6.2
✓ Peak Analysis & Results Code - See
Section 6.2
Figure 51: Edit Menu

✓ Inventory Checklist – The Inventory Checklist is designed to aid in keeping a regular tab on your
consumables inventory. Keeping an updated Inventory Checklist will assist in seamless monthly
inventory transitions. Addition of inventory items can be done by touching or clicking the ‘+’ button, and
deletions are done by touching or clicking the ‘-’ button. The Inventory Checklist is printable, and can be
quite helpful in monitoring inventory levels.
✓ By printing an inventory report a column report is generated with information on
Figure 52: Inventory Checklist Dialogue
the number of injection type carried out by the Operator.
6.5.3 View Menu
The View Menu & all of its corresponding options can be found below.
Figure 53: View Menu

✓ Batch – Allows the user to view results using the batch number as their means of reference. While
previous batches cannot be viewed during sample analysis samples from the current batch can be
reviewed while analysis is still ongoing.
Figure 55: Batch Icon - View Menu
Figure 54: View Batch Drop-Down Menu
✓ Search Results – Allows the user to search for results by using either a sample ID or the date in which
the
sample was run. Results can then be viewed, printed, or transmitted to an external device.
Figure 56: Search Results Dialogue

✓ Levey-Jennings Report – The Levey-Jennings Report catalogues your historical QC data for future
review. The Report is helpful in gaining a snapshot of how your instrument has been performing over
specified time periods, or for specific lots of QC Material. The data can be printed, or transferred to an
external device from this screen.
Figure 57: Levey-Jennings Plot

✓ Transmit Data to LIS – This menu allows the user to transmit data to a Laboratory Information
System. The Premier Hb9210TM transmits to LIS unidirectionally and bidirectionally using ASTM
protocols.
Figure 58:Transmit Data to LIS Dialogue
✓ Interface Language - The Operator is able to set up their Premier Hb9210TM in any of the following
languages: English, Brazilian Portuguese, Chinese, Danish, French, German, Italian, Latin-American
Spanish, Russian, Spanish (Spain) & Turkish.
Figure 59: Language Interface Selection Buttons

✓ Copy Files – The Copy Files menu allows the user to copy data to a desired location on the Premier
Hb9210TM PC or to an external device.
Figure 60: Copy Files Dialogue
✓ Activity Log – The Activity Log chronicles all user carried out actions executed on the instrument.
Figure 61: Activity Log

✓ Maintenance Log – The Maintenance Log allows the Operator to view all maintenance activities
logged in the
system via the Maintenance Log input icon in the Devices Menu.
Figure 62: Maintenance Log
6.5.4 Devices Menu
Figure 63: Devices Menu

The Devices Menu is primarily used by Field Service Engineers and should only be utilised by users
experienced in its use, or under the supervision of a Trinity Biotech, or local technical support
representative. The Devices Menu and all of its corresponding options can be found below.
✓ Pumps/Solvent Control – The Pumps button will bring up the Solvent Control screen. The flow rate
and gradient of Buffers A, B, and Wash can be chosen from this screen, or amount of purge time needed.
Figure 64: Devices Menu - Pumps Button
Figure 65: Devices Menu - Pumps Button - Solvent Control Dialogue
✓ Pumps/Purge Pumps – The Purge Pumps dialog box allows air bubbles to be purged from the bottles
through the pump heads. This dialog box appears whenever reagents Buffer A, Buffer B or WASH are
changed.
Figure 66: Devices Menu - Pumps Button - Purge Pumps Dialogue

✓ Oven – The Oven button allows the user to turn
the Oven Module on, off, or to reset the device.
Figure 67: Devices Menu - Oven Button
✓ Detector – The Detector button allows the user to Reset, Zero Output, and Start/Stop Data Acquisition.
Figure 68: Devices Menu - Detector Button

✓ Autosampler – A number of different functions can be performed via the Autosampler button. These
functions are listed below:
Figure 69: Devices Menu - Autosampler Button
- Reset – Resets the Autosampler, and is typically used for technical service purposes - Load Syringe –
Performs 1 Syringe aspiration to allow plunger to be disconnected, then pushed into glass barrel, then
syringe unscrewed at top. Installation performed in reverse, press RESET to complete syringe installation.
- Switch Valve – Switches the Injection Valve from Load to Inject. A troubleshooting utility designed to
aid
in clearing debris from the Injection Valve - Prime Diluter – Primes the dilution chamber where
patient whole blood samples are diluted and lysed for
analysis - Prime Rinse Station – Primes the rinse station - Cycle Racks – Cycles any racks present on
the sample transport through to the front of the Autosampler - Control Wheel Check – Checks to make
sure control wheel is in home position - Waste Pump – Allows Operators to turn on/off the Waste Pump
to purge waste from the system manually

✓ Barcode Reader – Turns the Barcode Reader on
or off.
Figure 70: Devices Menu - Barcode Reader Button
Figure 71: Devices Menu - Commander Button
✓ Maintenance Log – Allows operators log any maintenance activities that have been carried out on the
system.
✓ Commander – Should only be utilised by users experienced in its use, or under the supervision of a
Trinity Biotech, or local technical support representative. The Commander allows users to directly
command instrument to undertake certain tasks without going through the Software Program.

6.5.5 Help Menu
The Help Menu & all of its corresponding options can be found below.
Figure 72: Help Menu
✓ User Manual – Links the user to an electronic copy of the Premier Hb9210TM Operator’s Manual.
✓ Contact Info – Provides general contact info for Trinity Biotech Technical Service Representatives, as
well as empty fields for supplementary entry of helpful phone numbers.
Figure 73: Help Menu - Contact Info Dialogue

✓ About – The About Menu button provides the following information (appropriate for each GUI
software version):
• Software version
• Middleware version
• Column Injection Count
• System Injection Count
Figure 74: Help Menu - About Dialogue
6.5.6 Exit Application
The Exit Application button outputs can be found in Figure 75.
Figure 75: Exit Application

✓ Touching or clicking on the Exit Application button will end the user session of the Premier
Hb9210TM. The below screen will appear if this button is touched or clicked:
Note: The system must be inactive before exiting the program. Figure 76: Exit Application Confirmation
dialogue
6.6 Directory Structure

This software uses two main sub-directories for program locations and data archive,
“C:\PREMIER\GHB\DATA” and “C:\PREMIERARCH”. Programs used for running of this software and
data storage of recent batches of data are located in the “C:\PREMIER\GHB” sub-directory. Long term
archival storage of batch data is located in the “C:\PREMIERARCH\GHB” sub-directory. Note: all
batches except the newest 100 are automatically archived. Breakdown of each sub-directory and the type
of files they contain are as follows:
C:\PREMIER – System drivers and programs and files that are common to many versions of our software
platforms are located in this sub-directory.
C:\PREMIER\GHB – Programs and files that are specific to this version of assay are located in this
sub-directory.
C:\PREMIER\GHB\DATA – Recent data from the analysis of the samples are located in this
sub-directory.
C:\GHBARCH – Only contains sub-directories of the long-term archival storage of batch data.
C:\GHBARCH\2015 – Only contains sub-directories of each year’s batch data. The name of this example
sub- directory represents the 4-character year in which the data is from.
C:\GHBARCH\2015\20150103 through C:\GHBARCH\2015\20150408 – Examples of sub-directories
that contain each days batch data (January 03, 2015 through April 08, 2015). The name of this example
sub- directory represents the 4-character year, 2-character month, and 2-character day in which the data is
from.
The individual batches run on the selected date are contained in the folder.

6.7 Copying from the GHbArch Folder (the archive folder)
In order to be able to reprint or recalculate data older than the last 100 Batch Summary reports, the Batch
Summary Report file or the data files must be copied back into the folder at C:\Premier\GHb\Data.
From the desktop, open the My Computer icon to view the folders on the computer. Select Local Disk
(C:). Double-click on the GHbArch folder. You may need to scroll down the folder listing to find the
GHbArch folder.
To reprint the Batch Summary Report only, open the folder for the year in which the batch was run.
Select the folder with the date on which the batch was started; the first 4 digits represent the year, the next
two are the month, and the last two digits are the day date.
Within the date folder select the Batch Summary Report file (with the .BSR file extension) that
corresponds to the batch number to be reprinted. The first four digits represent the Batch Number. For
example, the file 0029.BSR is the Batch Summary Report for Batch 29. Highlight the BSR file name and
choose ‘Copy this file’ from the File and Folder Tasks listed on the left-hand side of the window. In the
Copy Items window select the folder Premier\GHb\Data and click the Copy button.
To reprint the batch including chromatograms, open the folder for the year in which the batch was run.
Select the folder with the date on which the batch was started; the first 4 digits represent the year, the next
two are the month, and the last two digits are the day date.
Within the date folder select the raw data files (with the .RAW file extension) that correspond to the batch
of chromatograms to be reprinted. The first four digits represent the Batch Number and the last three
digits are the individual chromatograms. For example, the file 0029001.RAW is the second
re-equilibration injection for Batch 29. Highlight the RAW file names for the entire batch and choose
‘Copy this file’ from the File and Folder Tasks listed on the left-hand side of the window. In the Copy
Items window select the folder Premier\GHb\Data and click the Copy button.
Data that is copied back into the Premier\GHb\Data folder is automatically deleted by the software to
maintain only the newest 100 Batch Summary Reports in the Data folder.
Also, if the Batch numbering is reset and batch numbers are being reused, the new chromatograms will
overwrite the older ones in the Data folder.

PREMIER Hb9210TM Results
& Interpretation
CHAPTER CONTENTS
✓ Auto-Verification ✓ QC Recommendations ✓ Report Interpretation

1 | Chapter 7 – Results & Interpretation Rev. 23
Chapter 7 – Results & Interpretation

7.1 Overview
At any given time that the system is ‘Running’, it generates two reports for each analysis cycle – an
On-Screen Report & a Printed/PDF Report. This chapter will review how to interpret these reports. All of
the following analysis sub-types will generate reports:
✓ Re-equilibrations ✓ QC/Controls Analysis ✓ Baseline Calibrations
✓ System Calibrations ✓ Patient Sample Analysis
One report can be viewed on-screen while the other is either printed or saved in PDF format (or if
selected, sent to LIS; this report will vary depending on local LIS systems). All data is then stored in the
directory on the C: drive of the Premier CPU Module for the purposes of data backup & storage. This
chapter will provide examples of both on-screen & printed reports, as well as recommendations for
interpretation.
7.2 Auto-Verification
The Premier Hb9210TM is designed with walk-away capability and does not require review of individual
reports – these features are defined as Auto-Verification.
The Auto-Verification feature of the Premier Hb9210TM is designed to assist with automatic review of
chromatography variables that an operator would normally check for, including:
✓ A stable re-equilibration baseline ✓ Acceptable peak area & valley heights ✓ Acceptable
pressure differences
✓ Acceptable retention times ✓ Acceptable Control/QC values
With multiple software checks in place, Auto Verification has been developed based on the following
platform benefits:
...Boronate Affinity Chemistry is a simplified separation method where only 2 peaks need to be
accurately quantified...
...a highly sensitive detector paired with advanced peak analysis software which gives an
accurate analysis of basic chromatogram quality...
Please note: the lab director has the responsibility to select and implement Auto-Verification rules and
processes that are appropriate for his or her lab. To use the Auto-Verification feature it is essential that
GLP recommendations are followed, specifically in regards to frequency of QC sample analysis. (see
Section 7.3 for QC recommendations)

7.2.1 Auto-Verification Features
There are two measures that must be followed if the Auto-Verification feature is to function in the manner
it was designed and in a manner in which it can be most effective to the laboratory.
1. Good (Clinical) Laboratory Practice (GLP/GCLP) QC guidelines are to be followed at all times.
2. Upon instrument installation, the Premier must be set-up by the laboratory administrator and the
appropriate
Trinity Biotech Technical Representative or Applications Specialist.
There are two measures that the Premier takes in order to ensure quality patient sample results.
Measure 1. Stable Baseline Confirmation
Each time a new column is installed on the instrument, a baseline calibration is run as part of the initial
calibration. The baseline ensures that any noise from the instrument is cancelled out and not interpreted as
part of final patient sample chromatography. Figure 1 below shows incorrect noise cancellation and how
it can be identified during the re-equilibration process.
Figure 1: Improper Cancellation of Baseline Noise from Final Chromatography
Noise from the pumps can be falsely interpreted by the detector as shifts in the chromatogram, and can
add unwanted peaks/area to the final result. The system will not allow the Operator to continue if:
1. A new column is installed – the system will not proceed to run the full System Calibration until a stable
baseline is detected.
2. During each patient sample analysis, four re-equilibration injections are performed to equilibrate the
column to analysis (in-run) conditions (temperature, flow, gradient). The fourth re-equilibration also
checks for baseline stability – it should be almost completely flat before injecting the first patient sample
in order to confirm and ensure that there is no unnecessary integration included in a final result
calculation.

Figure 2 provides an example of a baseline calibration. As emphasized by the noise circled on the screen,
it is not perfectly flat. Each baseline corresponds to a specific column. For this reason, an acceptable
baseline must be detected as part of the initial calibration each time a new column is installed. When the
Premier calibrates the baseline, it subtracts any noise from the system and will result in a flat and stable
baseline each time the re- equilibration injections are run.
Figure 2: Baseline Injection - Showing Typical System Noise
After a baseline is accepted, it results in a flat response from the detector when the re-equilibration
injections are run. In many cases, the first two re-equilibrations will exhibit some baseline noise, but this
is normal and due to the column conditioning itself for patient sample analysis.
The system has a built-in alert which checks the third and fourth re-equilibration for sufficient noise
cancellation. This means the following:
Four re-equilibration injections are run before any analysis cycle begins in order to ensure proper
conditions prior to reporting patient sample results.

✓ The system noise is being properly subtracted from any integration corresponding to the final patient
or QC
result ✓ The column is stable because the baseline has not shifted
In the event that the system does detect lingering system noise, or the baseline has shifted, the following
error message will appear and alert the user to re-run a baseline.
Figure 3: Baseline Limit Exceeded During 4th Re-Equilibration Alert Message
✓ An example of a flat fourth re-equilibration injection can be found in Figure 4.
Figure 4: Example of Flat Chromatogram Seen Upon Fourth Re-Equilibration

When viewing final chromatography, the original stored baseline can be seen as the green line on the
screen, while the actual detector response is seen in red.
Figure 5: Sample Run Showing Original Baseline in Green
Note in Figure 5 the original stored baseline in green and the final detector response in red. This green
trace is an indication of the system noise the Premier Hb9210TM is cancelling out to ensure optimal
analytical conditions.

Measure 2. Software Checks
The Premier Application Software has a feature entitled Peak Analysis & Results Codes which checks for
typical inconsistencies or irregularities in the final chromatography. This feature has been designed to
help the operator with interpretation of the chromatograms by ensuring that the correct result is being
generated. If any alarms are triggered, the system will provide an audible response which in turn alerts the
user that something has gone wrong. This feature is utilized with both patient sample runs and calibration
runs and has separate criteria for each scenario.
The Peak Analysis & Results Codes can be personalized to fit the QC policy of any laboratory, and
contain the following flexible preferences:
✓ Modifiable Results Codes can be adjusted within the manufacturer default ranges, not outside ✓
Frequency of occurrence before alert ✓ Whether or not to abort a run after ‘x’ amount of occurrences
The lab director has the ultimate responsibility to select and implement auto-verification rules and
processes that are appropriate for his or her lab.
The Peak Analysis & Results Codes for both Sample Run and Calibration Run are presented in Figure 6
& Figure 7 respectively, and are explained in-depth further on in the reading.
Although the Hb9210TM verification features in the Peak Analysis & Results Codes will sound an alarm
to alert the operator to any irregularities, it is still strongly recommended that the Operator reviews patient
results – even if baseline is stable & controls in-range.
Although the Peak Analysis & Results Codes are customizable, it is recommended that the manufacturer
defaults are only adjusted if advised to do so by a representative of Trinity Biotech or your local support
representative.

Figure 6: Peak Analysis & Results Codes - Patient Sample Runs
The different variables that the Samples Peak Analysis & Results Codes check for are defined as follows:
1. Total Peak Area – Measures the total area absorbed by the detector. 2. Height of Valley – Measures the
height of the valley between the two peaks. 3. Pressure Difference during Run (%) – Measures the
percentage pressure differences within a run. 4. Pressure during Analysis (bar) – Alerts the user of the
system exceeding the minimum or maximum allowable
pressure within a run. 5. First Peak Retention Times (minutes) – Alerts the operator of an
unacceptable first peak retention time. 6. Second Peak Retention Times (minutes) – Alerts the operator of
an unacceptable second peak retention
time. 7. Control I Result (range) – Alerts the operator of an unacceptable Control I result1. 8. Control
II Result (range) – Alerts the operator of an unacceptable Control I result1. 9. Value Result (linearity) –
Alerts the operator of a result detected by the system which falls out of the defined
range. 10. Not Within Normal Patient Range (user defined) – Flags low and high %HbA1c 11. Low
GHb Result (anaemic patients) – Flags a potentially anaemic patient.
1 The control range will shift based on the lot of QC material barcoded into the system.

Figure 7: Peak Analysis & Results Codes - Calibration Runs
The different variables that the Calibration Peak Analysis & Results Codes check for are defined as
follows:
1. Total Peak Area – Measures the total area absorbed by the detector. Area count limits are more
stringent for
Calibrator/QC materials. 2. Height of Valley – Measures the height of the valley between the two
peaks. 3. Pressure Difference during Run (%) – Measures the percentage pressure differences within a
run.. 4. Pressure during Analysis (bar) – Alerts the user of the system exceeding the minimum or
maximum
allowable pressure within a run. 5. First Peak Retention Times (minutes) – Alerts the operator of an
unacceptable first peak retention time. 6. Second Peak Retention Times (minutes) – Alerts the operator of
an unacceptable second peak retention
time. 7. Control I Result (range) – Alerts the operator of an unacceptable Control I result2. 8.
Control II Result (range) – Alerts the operator of an unacceptable Control II result2. 9. Does not apply to
Calibration Run 10. Does not apply to Calibration Run 11. Does not apply to Calibration Run 12.
Unacceptable Area Difference during Calibration (%) – Indicates significant differences in calibrator
values,
which would provide a poor calibration curve.
2 The control range will shift based on the lot of QC material barcoded into the system.

13. Calibration Slope Value – Ensures the calibration slope does not exceed a value of 2 – selecting the
checkbox will void calibrations which exceed this limit.
As is demonstrated in Figure 8 below, there are a number of points where key elements are confirmed by
the Peak Analysis & Results Codes in order for the chromatography to be considered acceptable. These
all help to assist the operator report the correct patient result. Also shown in green on the figure is the
system noise (e.g. from the reagent pumps) that is cancelled out of the final result.
Figure 8: Peak Analysis & Results Codes Points of Confirmation
7.2.2 Auto-Verification Features Conclusions
With multiple software checks in place paired with strict adherence to the QC recommendations made by
Trinity Biotech (based on GCLP & other key opinion leader recommendations), the feature is designed to
assist the operator with additional chromatography reviews. The software will alarm and alert the
Operator in case of any irregularities in the preset Peak Analysis & Results Codes parameters, which will
contribute to increased efficiency around the lab while maintaining confidence that the Premier will give a
precise and accurate result.

7.3 QC Recommendations
As part of good laboratory practices, operators should run controls and ensure values are within the ranges
stated on the package inserts. The Premier Hb9210TM allows for automated QC analysis at set intervals
in a batch, and Peak Analysis & Results Codes will alert operators if values fall outside of the stated
ranges. Frequently running QC samples assures the operator that the correct result is being generated. Of
the two Auto-Verification confirmations the system features, proper frequency of QC analysis is the most
important.
Trinity Biotech recommends that each level of control should be run at the beginning and end of each
analytical run, as defined by the laboratory, before reporting patient results.
Additionally, Trinity Biotech recommends that the chromatography from each QC/Control analysis be
reviewed by the operator for correctness prior to releasing patient results. The Auto-Verification features
have been designed to aid the Operator and help to reduce operator intervention. Auto-Verification acts as
a software-driven simulation of what an operator would normally check for in chromatography, however
it cannot fully replace a highly trained operator in regards to chromatography review and QC/Control
checks.
QC materials should be used in accordance with local, state, federal and accredited organizations. For
more information, please refer to your local guidelines for quality control regulations.

7.4 Report Interpretation
As mentioned, the system will generate 2 reports for each injection the system makes; a printed report &
an on- screen report for review. An example of the on-screen report is provided in Figure 9 while an
example of the printed report can be found in Figure 10.
7.4.1 On-Screen Report
The on-screen report is viewed on the Premier Touch-Screen Monitor each time any type of analysis is
carried out. The current sample being run is found on the top right of the window in real-time while
previous samples are shown on the larger main window for optional review.
Figure 9: On-Screen Report
7.4.2 Printed Report
The printed report can be generated in the PDF FactoryTM software featured in the Premier Application
Software or to a local/network printer. Figure 10 provides an example of a printed report.

Figure 10: Individual Report - Printout
7.4.3 Interpretation of Report
This section defines each element of the printed report (see Figure 10):
✓ “Batch xxx” indicates the batch number that has been assigned to the run by the computer. If the batch
is to
be reprinted, this number will be used to recall the batch from the hard drive.
✓ “Vial x:yyy” indicates which sample was injected. The “x” value tells which type of rack the sample
came from, W for Whole blood sample rack, H for Haemolysate sample rack and A for Anemic sample
rack. The “yyy” value tells which vial in the rack was injected.
✓ “abcdewxay” after the vial number is the patient ID. ID’s can be entered in three ways.
·
Automatically entered via the automated barcode reader
·
Key in names – the name will appear in the ID field.
·
Manually scan in Bar Code – the bar code will appear in the ID field.

✓ Date and Time the sample was injected.
✓ “Pressure = xx bar” – displays the pressure of the system at the point of injection. The “(xx to yy)”
numbers
show the extremes in the pressure readings during the sample run.
✓ Depending on reporting units selected, the values mMol HbA1c/mol Hb = yy, and/or HbA1c = x.x% –
are the
percentage of the glycated components in the sample.
✓ The chromatogram is a plot of the signal output from the detector. The first peak is the non-glycated
peak
and the second peak is the glycated peak.
✓ Retention time (RT) – the elapsed time between the start of the run and the apex of the peak.
✓ Total Area – the combined area of both peaks
✓ [FD4873564F865674] – this is a unique file identifier where the run data is stored.
✓ CONC = x.xx% is the area percent of the peak.
7.5 Other Reports

7.5.1 Header Page
The Operator has an option to print a Header Page at the beginning of each patient sample batch. The
option to print the Header Page can be enabled in the Run Parameters menu, as can be referenced in
Figure 11.
The Header Page contains very useful information, especially for Technical Support Teams in instances
of troubleshooting. The following information is contained in the Header Page:
✓ General Instrument ID ✓ Type of Analysis ✓ Operator Name ✓ Customer ✓ Department ✓
Address ✓ Location ✓ Date ✓ Batch # ✓ Lot Numbers (Reagents) ✓ Serial Numbers (Consumables)
✓ Calibration Details ✓ Date Report Printed

Figure 11: Print Header Page Option (Run Parameters Menu)
Most of the above information can be edited in the System Parameters menu, as shown in Error!
Reference source not found.. An example of a Premier Header Page can be found in Figure 13.
Figure 12: Header Page Editable Fields

Figure 13: Example Header Page

7.5.2 Batch Summary Report (BSR)
The Batch Summary Profile summarizes information from the individual reports in a convenient form for
review. The Batch Summary Profile is automatically generated at the finish of each batch run.
Dividing lines, or batch separators, can be added to the BSR in desired multiples to allow the Operator the
option of a more convenient method of counting down the BSR page if they need to find a result further
down in the report. To add them, tick the selection button in the Run Parameters dialogue.
Figure 14: Batch Summary Report (BSR)
Figure 15: BSR Separator Option

7.6 Chapter References
1. Ezzelle, J. et.al. Guidelines on Good Clinical Laboratory Practice.. J. Pharm Biomed Anal. 2008 January 7; 46(1):
18-
29. 2. College of American Pathologists (CAP). Laboratory General Checklist. 2007. 3. Clinical Laboratory
Improvement Amendments (CLIA) Equivalent Quality Control Procedures Brochure #4. 4. Statistical Quality
Control for Quantitative Measurement procedures: Principles and Definiftions; Approved Guideline 3rd
Edition C24-A3 Vol.26 No.25, 2006 5. ISO 15189 2012 Medical Laboratories, Requirements for Quality and
Competence 6. Basic QC Practices 3rd Edition, James O. Wetgard, 2010 7. Tietz Textbook of Clinical Chemistry
and Molecular Diagnostics, 2012

PREMIER Hb9210TM
Operator Maintenance
CHAPTER CONTENTS
✓ Changing Reagents ✓ Changing a Column ✓ Module Maintenance

Chapter 8 – Operator Maintenance
8.1 Introduction, Instrument & Component Overview
This chapter will contain an overview of routine maintenance procedures that an instrument Operator will
be expected to perform.
Maintenance checklists, that include suggested daily, weekly, and general procedures are included, and
may be incorporated into a laboratory logbook.
Each procedure will list required tools, required materials (if applicable), required time, and a procedure.
All of the tools that are required for the procedures in Chapter 8 are included with the Premier
Hb9210TM Startup Kit.
Figure 1: Premier Hb9210TM Operator Changing a Reagent Bottle
1 | Chapter 8 – Operator Maintenance Rev. 23

1. Do not activate the system if covers or access panels have been removed or damaged.
2. If moving parts are unavoidably exposed during normal use, be aware that any and all instructions
could result in injury, even if they are carried out by trained personnel. Only operators who have been
warned of the potential hazards and have received adequate training in carrying out the procedures in the
safest possible manner should attempt any/all procedures. Contact Trinity Biotech or your local support
representative for further information.
3. System pressures on an instrument can be up to 40 bar (580 psi) under rare circumstances. Due to the
very low volumes of reagent in the system, it is minimally dangerous. Contact Trinity Biotech or your
local support representative for further information.
8.1.1 Daily Maintenance
The Premier requires very little daily operator attention. The following are what Trinity Biotech
recommend be carried out on a daily basis:
✓ Performing regular activation & deactivations (Time Required: Automated)
- During activation, ensure that Buffer A & Buffer B debubblers are full (for serial numbers 200733 or
earlier
– 200734 and later feature Auto-debubblers) (Time Required: 1 minute) ✓ Wiping down
instrument surfaces & making sure that lab area is clean & tidy (Time Required: 1 minute) ✓ Ensuring
that reference material is stored appropriately when QC dilutions are prepared (Time Required: 1
minute)
Before carrying out any maintenance activities, please make note of the points highlighted below.

8.1.2 Weekly Maintenance
✓ Changing of the Pump Module recirculating purified water
shown in Figure 2 (Time Required: 5 minutes) ✓ Powering off system over weekends or when
instrument
will go unused for longer periods of time (2-7 days) (Time Required: 1 minute) ✓ Cleaning the probe
needle guide black plastic insert
8.1.3 Periodic Maintenance ✓ Following column change instructions (on-screen)
- Changing Frits (Time Required: 5 minutes) - Disposing of old column in biohazardous waste bin
(Time Required: 1 minute) ✓ Following reagent change instructions (on screen) (Time
Required: Variable)
Figure 2: Pump Module Recirculating
Water ✓ Proper instrument care if going unused for longer
than a week - Perform a deactivation (Time Required:
Automated) - Remove column, replace end-caps & place in a 2- 8°C refrigerator – install Column
Placeholder, 09- 43-1804, shown in Figure 3 (Time Required: 5 minutes)
8.1.4 Preventative Maintenance
✓ Scheduled by technical service representatives
Figure 3: Column Placeholder

8.2 Changing the Reagents
Required Tools: None
Required Materials: Applicable (empty) bottle of Premier reagent
Required Time: 5 minutes (2 minutes to change reagent bottle, 3 minutes to prime system)
Reagent levels can be checked from the Main Assay screen, or before running any type of analysis.
Figure 4: Reagent Levels Quick Touch Button
Note: There are 3 different operator notifications in place on the Premier which alert the operator of
reagent levels and of the relative need to change the appropriate bottle(s) at a given time in the near
future.
The first, as shown in Figure 5, is the estimated number of injections to run count in the level sense
dialogue. This provides an estimate of remaining injections given current reagent levels in place on the
Premier Hb9210TM. This is a reflection of the reagent with the lowest remaining volume and is not a
reflection of column injections remaining.
Figure 5: Reagent Levels

Reminder 1. The reagent level will turn Yellow. Approximately 50 patient samples remain before the
system will deactivate. The system will audibly notify the operator – the operator will need to manually
deactivate the audible alarm.
Figure 6: Reminder 1 - Audible Alarm - Reagent Levels Low
Reminder 2. The reagent level will turn Red. The system will not allow any further samples to be run. The
status bar will reveal a 15 minute timer until the system automatically deactivates. The system will
audibly notify the operator. The operator must change the reagent.
Figure 7: Reminder 2 (Final Reminder) - Audible Alarm - Reagent Levels Low
As reagent levels decrease through use, Operators will be presented with reminders to alert them of the
levels remaining in the bottles. This ensures reagents are changed in a timely manner so as to avoid the
introduction of air into the system via pumping from empty reagent bottles.

At this point, the Premier will display an on-screen message to change the Reagent bottle. The on-screen
message reads as follows:
Premier Hb9210TM Reagent Bottle Change Instructions:
1. With the system flow at 0.0 mL/minute1 (at either Standby or Inactive) and after the new reagent bottle
barcode has been scanned, locate the empty bottle to be removed from the analyzer.
2. Unscrew reagent cap and remove lines from bottle. CAUTION – Do not pull on reagent tubes – this
can
cause the reagent liquid level sensing system to be inaccurate and/or damage the tubing.
3. With clean tissue, wipe the bottle tubes dry and prevent tubing tips from touching the reagent tray or
other
surfaces.
4. Remove empty bottle from system.
5. Remove cap from the new bottle of reagent. Remove foil seal from bottle. Place bottle in reagent tray.
6. Insert reagent lines into new bottle and tighten reagent cap. Be sure the tubing lines are not twisted or
crossed from the instrument to the bottle top.
7. Repeat steps 1-6 for any additional reagents to be replaced.
8. Allow system to Auto-Prime any/all reagents that were changed, according to dialog box(es).
9. When controls are within range, sample analysis can proceed. You do not need to calibrate unless the
controls are out.
1 The Flow can be set to 0.0 mL/min by pushing Ctrl + S from the Main Assay Screen, or from Devices Menu →
Pumps → Solvent System.

Procedure:
1. Obtain new bottle of reagent in need of changing. From the Lot Numbers Dialogue, scan in the new
reagent barcode using the barcode gun.
Figure 8: Buffer Barcode Entry (from Lot Numbers Dialogue)
Figure 9: Purge Pumps Control Dialogue
2. Follow the on-screen instructions to change the reagent bottle.
3. The system will automatically alert the Operator to prime. Prime the system for three minutes.
4. Lift the Pump Door & manually check to assure that the debubblers are primed.2
5. Begin to run samples once
the system is done priming.
The locations of the reagents on the system can be found in Figure 10 below.
2 To prime the manual debubblers, loosen the top valve and allow reagent level to fill chamber. Only applicable for
serial numbers 200733 and earlier.

Figure 10: Physical Locations of the Premier Hb9210 Reagents (XL500 and XL1000 test packs)
Reagent Side Trays can also be fitted to your Premier Hb9210TM Analyzer for use with the XL3000 test
packs. Please contact Trinity Biotech or your local distributor how to acquire the Side Trays.
Figure 11: Diluent Left Reagent Side Tray (Left), Buffer Right Reagent Side Tray (Right)

8.2.1 Priming the Pumps
It will be necessary to prime the buffers any time the solvent flow path is opened up in the pump (i.e. to
replace pump seals or check valve), if bubbles are present in the solvent lines, or if the pressure
fluctuation is unacceptable. This can all be done through the software by following this simple procedure:
Required Materials: On-board reagents
Required Tools: None
Required Time: 5 minutes
Procedure:
1. From the Main Menu screen, select Devices, then Pumps, followed by Purge Pumps.
2. Set the purge time to at least 2 minutes.
3. Once complete, check system for presence of air bubbles or unprimed pump.
8.2.2 Priming the Piston Rinsing Chamber with Purified Water
The piston wash is Purified Water (purified, filtered, DI, and/or distilled) that is used to continually rinse
the piston when the pump is running. This purified water recycles through both pumps and should be
changed weekly. Prime (lack of large air bubbles) of the Purified Water lines to the pump heads should be
checked on a daily basis. NOTE: It is natural for some small air bubbles to move in the lines.
Materials Required: Purified Water (purified, filtered, DI, and/or distilled)
Tools Required: Priming Syringe
Procedure:
1. Ensure system status is in standby.
2. In the Application Software, select Devices and then Pumps/Solvent Control. Set the flow to 1.00
ml/min,
50% Pump B. Press set.
3. Place the priming syringe in the priming tee connection located to the right of the pump on one of the 2
recirculating pump head rinse lines. Turn the arm of the priming tee down (away from bottle) and pull
back on the plunger of the syringe. Turn arm of priming tee towards syringe connection point. Remove
syringe
Be sure that debubblers are fully primed as per daily maintenance requirements; both during and after
purging. See Section 9.1.2 for more specific instructions on how to prime the manual debubblers.

and expel purified water. Repeat until all of the air has been removed from the solvent line between the
Purified Water bottle and the priming valve.
4. Place the priming syringe in the same priming tee, but turn the arm of the priming tee up. Pull back on
the plunger of the syringe. Turn the arm of the priming tee towards the bottle. Press on syringe to push
purified water through the pump heads. Repeat until all of the air has been removed from pump head
wash lines and back to the bottle.
8.2.3 Priming the Dilutor (Premier Hb9210TM DIL Reagent)
The Premier Hb9210TM DIL Reagent is used by the syringe module to aspirate the samples as well as to
help clean the septum-piercing probe. Air in the line between the Premier Hb9210TM DIL Reagent bottle
and the septum piercing probe will cause inaccurate dilutions. Priming the Premier Hb9210TM DIL
Reagent is accomplished completely through software.
Materials Required: Premier Hb9210TM DIL Reagent, 3.8L, 01-03-0097
Tools Required: None
Procedure:
1. Confirm the inlet line from the tan valve is completely submerged in the Premier Hb9210TM DIL
Reagent.
2. Confirm both ends of the Dilutor syringe are tightened securely by hand.
3. Confirm the inlet and transfer tubing is securely attached to the syringe manifold (tan block above the
syringe).
4. From the Application Software go to Devices, Autosampler and click on Prime Dilutor.
5. A Dialog Box will appear, the Autosampler will perform a homing sequence and the septum-piercing
probe
will move over the rinse station. The syringe will then begin to cycle at 100% of its capacity.
6. When all air bubbles have been removed from the inlet and transfer lines click on the OK button to stop
the
cycling of the syringe.
8.2.4 Priming the Active Rinse Station (ARS)
Premier Hb9210TM DIL Reagent is used for the Active Rinse Station (ARS) reagent. The ARS is used by
the probe module to facilitate dilution of the whole blood samples and to help clean the rinse station and
septum- piercing probe. If the ARS is not pumping the Premier Hb9210TM DIL Reagent, the result could
be over concentrated whole blood injections and/or carryover. Priming the ARS is normally accomplished
through the software.
Materials Required: Premier Hb9210TM DIL Reagent, 01-03-0097

Procedure:
1. Confirm the clear tubing from the ARS is completely submerged into the bottle of Premier Hb9210TM
DIL
Reagent.
2. From the Application Software go to Devices, Injector and click on Prime Rinse Station Pump.
3. A Dialog Box will appear and the ARS pump will turn on. Leave pump on until a small amount of
Premier
Hb9210TM DIL Reagent appears to flow from the top of the tan insert of the rinse station.
4. Click on OK in the dialog box to turn the pump off.
8.3 Changing the Column

Materials Required: Column, Pre-Injection Valve Frit, Pre-Column Frit
Tools Required: no tools are required
Required Time: 40 minutes (< 5 minutes required for Operator)
Note: There are 3 different operator notifications in place on the Premier which alert the operator how
many injections are remaining on the column.
Reminder 1. The Run Baseline, Run Calibration & Run Samples windows will all have an ‘injections
remaining’ notification at the top left of the dialogue.
Be very careful when changing the column. If the system hasn’t undergone a full deactivation before
changing the column, the oven and column will be very hot to the touch.

Figure 12: Reminder 1 - Column Injections Remaining (from Analysis Confirmation Dialogues)
Reminder 2. An Audible Alarm will sound until manually overridden by the operator.
Figure 13: Reminder 2 - Audible Alarm - 25 Column Injections Remaining
Reminder 3. When there are 0 injections remaining on the column, a new column will need to be
installed.

Figure 14: Reminder 3 (Final Reminder) - Audible Alarm - No Further Column Injections Remaining
Procedure: The following procedure requires additional requirements. The Laboratory Administrator will
be fully trained on the procedure upon instrument installation.
Additional Requirements. In order to change the column, the operator will need to do the following:
✓ Scan in column lot number ✓ Load QC & Calibrators ✓ Change Frits
✓ Activate System ✓ Run Baseline ✓ Run Calibration
1. Scan in new column lot number. To do this, navigate to the Lot Numbers window. From the Main
Menu, select the Edit Menu.
Figure 15: Edit Menu Button (from the Main Menu)

2. From the Edit Menu, select the Lot
Numbers button.
Figure 16: Lot Numbers Button (from the Edit Menu)
Figure 17: Lot Numbers Entry Dialogue - Column Tab
3. Select the ‘Column’ tab on the top of the dialogue. Touch the Scan Barcode button and scan in the
column barcode using the handheld barcode scanning device.
4. Once the new barcode has been scanned into the system, all of the physical changes can be made. The
physical column change instructions are provided in an on-screen message:

Figure 18: On-Screen column Change Instructions
5. After changing the column, be sure to change the pre-column & pre-injection valve frits before
activating the
system. Instructions for this can be found in section 8.3.1 below.
6. When the column has been physically installed & the frits have been successfully changed, the operator
will now need to Activate & Run Calibration before running patient samples. This can all be done
automatically through the Automated Operator Assistant.
7. Select the Run Samples dialogue from the Main Assay Screen or the Operation Menu. In the top right
of the Run Samples dialogue, enable the boxes in the Automated Operator Assistant to Activate & Run
Calibration (shown in Figure 19).
8. Load the QC material before enabling the Automated Operator Assistant. Begin the Run. The system
will automatically Activate, Run Baseline, Run Calibration & Run Samples.
Figure 19: Automated Operator Assistant Selection Menu

8.3.1 Changing the Frits
When changing the column, it is required to change the Pre-Injection Valve & Pre-Column frits. One of
each frit is included with each column. Procedures for how to change the frits are as follows.
Changing the 75μm Pre-Injection Valve Frit 09-71-1905
Materials Required: 75μm Frit – Part Number: 09-71-1905
Required Time: 2 Minutes
Procedure:
1. Locate the Injection Port-to-Injection Valve tubing (09-41-1621) – it is the orange PEEK tubing going
from
the Probe Module to Position 2 of the Injection Valve Module (not the clear waste tubing).
Figure 20: Locate the Pre-Injection Valve Frit Housing
The Premier Hb9210TM utilizes two types of Frit/Sample Pre-filter – 2μm Pre- Column and 75μm
Pre-Injection Valve

2. Loosen & remove the white fitting from sample pre-filter housing, which holds the 75-μm sample
pre-filter.
The housing is the tan fitting attached at Position 2 of the injection valve.
Figure 21: Loosen the White Frit Housing Fitting
3. Remove the tan frit housing/fitting from the Injection Valve. The 75 μm frit will still be in the tan
finger-tight
housing.
Figure 22: Remove the Tan Frit Housing from the Injection Valve

4. Remove the 75μm frit from the tan housing. Do this by drying the inside of the housing with a soft
tissue
and gently tapping the fitting on a hard surface until the frit falls out.
5. Discard old frit in proper biohazard disposal container. Install a new 75μm frit.
Figure 23: Newly Installed Pre-Injection Valve Frit (Colourless side Facing Upwards)
Reinstall Frit Housing Fitting, & Injection Port-to-Injection Valve Tubing in reverse order.
There are two sides to the 75μm frit – one is white, the other is colourless. Be sure to have the white side
of the 75μm frit facing downward, and the colourless side facing up. Failure to position correctly will
reduce filtering efficiency and can affect sample injection volumes.
✓ When re-installing the orange tubing, first loosely install the orange tubing
& white finger-tight back into the tan finger-tight frit housing. ✓ Do not tighten down – only loosely
install. ✓ Next, tighten down the tan finger-tight housing into Position 1 of the
injection valve. The orange tubing & white finger-tight was not tightened down so as to allow it the
ability to turn freely as the tan finger-tight was installed. ✓ Once the tan finger-tight is installed back into
Position 1 of the Injection Valve, the orange tubing & white finger-tight fitting can be tightened into
place.

Changing the 2μm (Pre-Column) Frit (03-11-0056)
Required Materials: 2μm Frit – Part Number: 03-11-0056
Required Tools: 2 Wrenches (capable of 7/16” and/or 11mm) – unless using 2μm Finger Tight Frit
Housing, PN 09-33-3205 – in which case no tools are required
Required Time: 2 Minutes
Procedure:The column pre-filter is located on the tubing between Position 6 of the Injection Valve
Module and the column pre-heating coil tubing of the Oven Module. It is set in a stainless steel frit
housing, as shown in Figure 24.
Figure 24: 2μm Pre-Column Frit Housing Orientation
Note: To change the 2μm Frit if using the Finger Tight Frit Housing, simply loosen the two aluminium
handles by hand as demonstrated in Figure 25.
Figure 25: Opening the 2μm Finger Tight Frit Housing

Remove and replace the 2μm Frit. Tighten by hand until you feel that the surfaces of each side of the
housing have met.
Hand-tighten two ‘notches’ further on the housing using the reference ‘notch’ on the handle as your
guide. See Figure 26 below for an example of tightening ‘two notches further’.
Figure 26: Instructions for Changing the 2μm Frit
8.4 Detector Maintenance

8.4.1 Removing Air Bubbles from the Flow Cell
Air bubbles can become trapped in the flow cell causing poor baseline and erratic results. Once the
system has been running it is unlikely that an air bubble will get trapped unless the system has lost prime
or the flow system has been opened up. The following is a procedure for removing trapped air bubbles
from the flow cell.
Materials Required: None
Tools Required: Column Bypass, Part Number 09-41-1704
Procedure:
1. In the Application Software, select Devices and then Pumps/Solvent System.
2. Set the flow to zero and allow the pressure to come down to less than 10 Bar.
Two ‘Notches’
Reference ‘Notch’

3. Remove the column from the system and install a column bypass union (this is a metal zero dead
volume
union that was included with the tool kit)
4. In the Solvent Control dialog box set the flow to 100% A and 3.0 ml/min and press Set. This will allow
a
high reagent flow through the flow cell and may flush any air bubbles from the flow cell.
5. Allow this flow to pass through the flow cell for at least 5 minutes.
6. In the Application Software, select Devices and then Detector and select Zero analog output.
7. Then select Devices, Detector and select Acquire data.
8. The output from the detector will be monitored for 50-seconds and is represented with the horizontal
blue
line on the screen. The baseline should be flat.
9. If the baseline appears to be flat set the flow to zero and reinstall the column.
8.5 Pump Maintenance

There are no components of the Pump that are user replaceable. Wear items are replaced on a periodic
cycle during preventive maintenance visits by Trinity Biotech Technical Service Representatives or your
local distributor.
8.6 Sample Transport, Syringe Module and Probe Module Maintenance

There are no components of the Sample Transport and the Probe Module that are user replaceable. Wear
items are replaced on a periodic cycle during preventive maintenance visits by Trinity Biotech Technical
Service Representatives or your local distributor.
Many of the components on the Sample Transport and the Probe Module injector come in direct contact
with the blood samples. The Premier Hb9210TM should be run through a shutdown cycle and wiped
down with disinfectant before removing or working on either of these units. Proper personal protective
equipment must be worn and proper precautions should be taken when working on either of these units.

PREMIER Hb9210TM
Operator Troubleshooting
CHAPTER CONTENTS
✓ Hardware Error Codes ✓ Peak Analysis & Result
Codes ✓ General Chromatography
Troubleshooting ✓ Contacting Tech Support

Chapter 9 – Operator Troubleshooting
9.1 General Troubleshooting
The Premier Hb9210TM auto-verification features have been designed to alert operators with alarm and
error codes to any potentially poor chromatography. This chapter outlines problems that can lead to poor
chromatography and the suggested remedies that may assist in correcting such problems.
If a resolution is not obtained within a reasonable amount of time, call your Technical Support
Representative. Basic preventative maintenance and care is the key to a system that runs well.
Contact your Trinity Biotech Technical Representative:

US/Canada/Puerto Rico Customers: Trinity Biotech USA 2823 Girts Road Jamestown, NY
14701 Tel: 1-800-325-3424 [email protected]
International Customers: Trinity Biotech IDA Business Park Bray, Co. Wicklow, Ireland Tel: +
353 1 276 9898 Fax: + 353 1 276 9888 [email protected]
www.trinitybiotech.com
9.1.1 Aligning the Touch Screen Monitor
Purpose: If the Touch Screen Monitor becomes unresponsive or imprecise to touch, it is occasionally
required to calibrate its alignment.
Time Required: 1 minute
1 | Chapter 9 – Operator Troubleshooting Rev. 23

Procedure:
1. Select the ELO icon from the Windows Taskbar
Menu, as highlighted in Figure 1.
Figure 1: ELO Touch Screen Alignment Icon in the Windows Taskbar
2. Select the Align button from the ELO dialogue,
as shown in Figure 2.
Figure 2: Alignment Button Selection

3. Touch the targets positioned
around the screen.
Figure 3: Positioning the Touch Screen
4. Drag your finger around the Touch Screen and ensure that the cursor follows your finger. If the cursor
follows your finger, select the green check mark to complete the alignment. If the cursor does not follow
your finger, select the blue return arrow on the right to return to Step 3.
Figure 4: Alignment Confirmation
9.1.2 Priming the Manual Debubblers
Purpose: If a system is being newly installed, maintenance performed, the system loses prime, reagent
bottles are changed, or any air is somehow being introduced into the system upstream from the Pump
Heads, the debubblers will trap air before it reaches the Pump Heads and will need to be primed.
Time Required: 1 minute
Manual debubblers are only applicable for Premier Hb9210TM serial numbers 200733 and earlier,
instruments with serial number 200734 and higher feature automatic debubblers

Procedure:
1. Lift the Pump Door.
2. Locate the Debubblers.
3. Turn the top valve counter-clockwise to purge. The reagent level will fill to the top. Turn clockwise to
tighten and close valve.
Figure 5: Priming Debubblers
If the debubblers are losing prime on a regular basis, check upstream from the debubblers for possible
points of entry of air/leaks.

9.2 Hardware Error Codes
This section will guide operators through the troubleshooting steps to be taken if any hardware error
codes or issues are observed. Following the flow charts from left to right the steps are as follows:
1. Error Code
2. Potential Cause
3. Resolution
If these steps do not resolve the error, or you observe any other hardware error codes, you should contact
a member of the Trinity Biotech Technical Team, or your local distributor immediately.
9.2.1 Data Handling System
The data handling system consists of a CPU, colour touch screen monitor, printer, and a handheld barcode
gun combined with the Premier Hb9210TM Application Software. The steps below can help operators
resolve some issues that may arise in the course of daily use. If you require information on your specific
computer, please contact Trinity Biotech or your local support representative.
Figure 6: Premier Hb9210TM Data Handling System (with covers removed to show CPU module)

AP3 Cannot Communicate with Middleware
“Out of Range” Error on the Monitor
Blue Screen on Startup, won’t boot windows.
Touch screen Unresponsive/Lost Calibration

9.2.2 Autosampler System
The Autosampler System consists of a Sample Transport Module, the Probe Module and Injection Valve
Module.
Faulty Keyboard Track-Pad
Device plugged into USB adapter hub not working properly
AP38 Run stopped because system timed-out looking for tubes or racks

AP114 Run aborted. Zero calibration result
AP139 Rub aborted. Consecutive duplicate vial barcodes detected
AS120-AS124 Yokes not in position to run conveyor

The error codes noted above share a common resolution pathway as shown in the flow chart below:
AS134 Indexing Error while moving the yoke
AS125 Cal Wheel not in position, AS126 Probe homing error, IV122 Assay time exceeded, IV119
Position error unable to obtain commanded position, IV165 Module uninitialized

SY2 Syringe in/out mismatch
Syringe module making clicking sound
Fluid dripping under the injection valve

9.2.3 Pump Module
Probe actuator making excessive noise
AP120 Communication Error
AP130 Communication Timeout

9.2.4 Oven Module
Premier Hb9210TM Oven Module performance specifications:
✓ Temperature Range: 30-70°C ✓ Temperature Stability: ±0.2°C ✓ Temperature Repeatability: ±0.5°C
✓ Temperature Accuracy: ±0.5°C ✓ Safety Cutout: 70°C
AP11 Temperature limit exceeded
9.2.5 Detector Module
AP138 Baseline out of range. Re-run baseline

9.2.6 Waste Pump
The automatic Waste Pump hardware is located under the Rear Lower Panel of the Premier Hb9210TM.
System waste leaking under the instrument

9.3 Peak Analysis & Results Codes
The Premier Hb9210TM has several safety codes – both for patient sample runs and for calibration runs –
designed to alert the operator of abnormalities within the reported chromatography results. However, not
all abnormalities can be quantitatively assessed by the software due to the wide range of patient results
experienced in a normal laboratory environment, so operators must be able to visually tell the difference
between acceptable and abnormal chromatograms (refer to section 9.4).
9.3.1 Patient Sample Peak Analysis & Results Codes
Figure 7: Patient Sample Peak Analysis & Results Codes
Although the Peak Analysis & Results Codes are customizable, it is recommended that the manufacturer
defaults are only adjusted if advised to do so by a representative of Trinity Biotech or your local support
representative.

Code 1: Unacceptable Total Peak Area
- Less than 250,000 total peak area
1. No obvious instrumentation problems: samples possibly too dilute – either being over-diluted by the
analyzer or
not enough sample in the patient sample tube. a. Check sample volume. Must have a minimum of 0.5mL
of hemolysate or 0.5mL of packed red blood cells
to sample correctly. b. Check the dilution if making hemolysate and verify that it is the proper
dilution ratio (1:150). 2. Air leaks: system erratically sampling due to air leaking into the sampling circuit.
If this is happening, air
bubbles should be visible in the tube connecting the syringe module to the probe. a. Tighten fittings on
Syringe Valve Manifold. b. Tighten Dilutor syringe (top and bottom). c. Tighten fitting at probe top. 3.
Diluent low: check to ensure that physical reagent levels match software reagent levels. 4. Plugged
Pre-Injection Valve Frit: 75μm frit on the injection circuit is plugged.
a. Replace the 75μm frit
- Greater than 2M total peak area
1. Whole Blood sample placed in an ‘H’ (hemolysate) rack.
a. If there was accidentally a whole blood sample placed on a hemolysate rack, stop analysis immediately.
Run the system at 50% Reagent A/50% Reagent B for 30 minutes at 1.00 mL/min to flush protein off the
column. Run baselines or blank injections after 30 minutes to test for residual material left over on the
column. 2. Sample dilution parameters adjusted incorrectly.
a. Contact Trinity Biotech technical service representative. Sample dilution parameters can only be
adjusted
by a trained Field Service Engineer.
Although the Hb9210TM has auto-verification features built into the Peak Analysis & Results Codes, it is
still strongly recommended that the Operator review each patient result.


Code 2: Unacceptable Height of Valley
- Valley height greater than 1,000
1. Loss of Prime: Air in the HPLC system.
a. Check for leaks. 2. Void Volume in the HPLC system. Void volume present in one of the critical
junctions in the HPLC system.
a. Check 2μm frit housing to confirm that it is tightened properly. b. Check column to make sure it is
tightened properly
Code 3: Unacceptable Pressure Difference During Sample Analysis (%)
- Unacceptable maximum pressure difference between pumps A & B during analysis
a. Check physical reagent levels, confirm that bottles have enough reagent and tubing is reaching the
reagent
levels. b. Check for leaks. c. Check debubblers, make sure the air is bled out of them. Remember to
check for air in debubblers with every change of reagent A or B and during activation as per Operator
Daily Maintenance recommendations. d. From Main Menu, go to Devices → Pumps → Purge Pump.
Turn the Purge Pump on for 2 minutes. Confirm
that the A and B reagent supply lines are primed completely. e. Remove the column and install bypass
union. From Devices → Pumps → Solvent Control, run both A and B
pumps at 50% each with reagents A and B selected (not wash) for 5 minutes. f. Remove bypass
union, install analytical column and activate the system. g. Run a Baseline and confirm that there is no
significant pressure drop within the run. NOTE: The first 1/3 of the screen is when the A pump is
pumping, the middle third of the screen is where the B pump is pumping and the last third of the screen is
when the A pump is pumping again. Lower pressure at the beginning, middle or end of the chromatogram
can indicate which pump has lost prime.

Code 4: Unacceptable Total Pressure during Run (bar)
- Unacceptable minimum system pressure during analysis - Unacceptable minimum-maximum pressure
difference between pumps A & B during analysis
a. Check reagent level, confirm that bottles have enough reagent and tubing is reaching the reagent levels.
b. Check debubblers, make sure the air is bled out of them. Remember to check for air in debubblers with
every change of reagent A or B. c. From Main Menu, go to Devices → Pumps → Purge Pump. Turn
the Purge Pump on for 2 minutes. Confirm
that the A and B reagent supply lines are primed completely. d. Remove the column and install
bypass union. From Devices → Pumps → Solvent Control, run both A and B
pumps at 50% each, 2ml/min, with reagents A and B selected (not wash) for 5 minutes. e. Remove
bypass union, install analytical column and activate the system. f. Run a Baseline and confirm that there is
no significant pressure drop within the run. NOTE: The first 1/3 of the screen is when the A pump is
when the A pump is pumping again. Lower pressure at the beginning, middle or end of the chromatogram
can indicate which pump has lost prime. 2. Leak in System:
a. Check tubing connections for leaks and tighten. 3. System pressure too high: plugged 2μm frit or
column
a. Change 2μm frit with every column change.

Code 5: Unacceptable First Peak Retention Time (minutes)
- The non-glycated peak did not elute in the acceptable peak window
a. Check reagent level, confirm that bottles have enough reagent. b. Check for leaks. c. Check debubblers,
make sure the air is bled out of them. Remember to check for air in debubblers with
every change of reagent A or B. d. From Main Menu, go to Devices → Pumps → Purge Pump. Turn
pumps at 50% each, 2ml/min, with reagents A and B selected (not wash) for 5 minutes. f. Remove
bypass union, install analytical column and activate the system. g. Run a Baseline and confirm that there
is no significant pressure drop within the run. 2. Void Volume in the HPLC system. Void volume present
in one of the critical junctions in the HPLC system.
tightened properly
Code 6: Unacceptable Second Peak Retention Time (minutes)
- The glycated peak did not elute in the acceptable peak window
every change of reagent A or B. d. From Main Menu, go to Devices, Pumps, Purge Pump. Turn the
Purge Pump on for 2 minutes. Confirm
tightened properly

Code 7: Unacceptable Control I Result
- Control level I material is not within the range stated on the package insert.
Note: If the chromatography is abnormal, refer to section 9.4 before performing the following steps.
1. Calibrate system. See Operator’s Manual section 6.3.2. 2. Incorrect dilution. Prepare new control
samples with the correct dilution ratio listed on the pack insert. 3. Reagent debubblers full of air. Bleed
the air out of the debubblers. 4. Control material. For lyophilized QC material, reconstituted QC material
and haemolysate dilutions of reconstituted QC material, ensure the correct storage, reconstitution and
expiration date, according to package insert(s).
Code 8: Unacceptable Control II Result
- Control level II material is not within the range stated on the package insert.
Refer to Code 7: Unacceptable Control I Result for appropriate Code 8 troubleshooting steps.

Code 9: Unacceptable Value Result
- Result of analysis is beyond the system linearity range
1. Rerun Sample. 2. Calibrate system. See sections 6.3.2. 3. Perform Linearity Evaluation.
Code 10: Not within Normal Patient Range
- Result of analysis is outside of the operator-defined normal range
Note: This code is not a system error. This code simply reminds the operator that the result is outside of
the normal range (for the reporting units the system was calibrated in). For mMol HbA1c/mol Hb
reporting, the IFCC has established 30-40 mMol HbA1c/mol Hb as their normal range. For %HbA1c
reporting, the NGSP has established 4.0-6.0%HbA1c as their normal range.
Code 11: Low Glycohaemoglobin Result
- Result of analysis is below the operator-defined limit for normal HbA1c levels.
Note: This code is not a system error. This code reminds the operator that the result is below the
established limit (by the manufacturer or the laboratory administration) for patients with normal %HbA1c
results. Values below normal %HbA1c limits could indicate abnormal haematology.

9.3.2 Calibration Run Peak Analysis & Results Codes
Figure 8: Calibration Peak Analysis & Results Codes
- Less than 380,000 total peak area
1. Cal/QC samples possibly too dilute.
a. Check sample volume. Must have a minimum of 0.5mL of Cal/QC material in the vial. b. Check the
dilution and verify that it is the proper dilution ratio specified in the Operator’s Manual and
Cal/QC Package Insert. 2. Air leaks: system erratically sampling due to air leaking into the
sampling circuit. If this is happening, air
bubbles should be visible in the tube connecting the syringe module to the probe. a. Tighten fittings on
Syringe Valve Manifold. b. Tighten Dilutor syringe (top and bottom). c. Tighten fitting at probe top. d.
Diluent low – check to ensure that physical reagent levels match software reagent levels. 3. Plugged
Pre-Injection Valve Frit: 75μm frit on the injection circuit is plugged.
a. Replace the 75μm frit

- Greater than 1.5M total peak area
1. Cal/QC re-constituted improperly
a. Re-constitute new Cal/QC material as per the Package Insert. 2. Cal/QC diluted improperly
a. Prepare new dilutions of Cal/QC material. Re-run calibration.
Code 2: Unacceptable Height of Valley
- Valley height greater than 1,000
a. Check for leaks. 2. Void Volume in the HPLC system. Void volume present in one of the critical
junctions in the HPLC system.
tightened properly

Code 3: Unacceptable Pressure Difference during Sample Analysis (%)
- Unacceptable maximum pressure difference between pumps A & B during analysis
a. Check physical reagent levels, confirm that bottles have enough reagent and tubing is reaching the
reagent
levels. b. Check for leaks. c. Check debubblers, make sure the air is bled out of them. Remember to
check for air in debubblers with every change of reagent A or B and during activation as per Operator
Daily Maintenance recommendations. d. From Main Menu, go to Devices → Pumps → Purge Pump.
Turn the Purge Pump on for 2 minutes. Confirm
pumps at 50% each, with reagents A and B selected (not wash) for 5 minutes. f. Remove bypass
union, install analytical column and activate the system. g. Run a Baseline and confirm that there is no
significant pressure drop within the run. NOTE: The first 1/3 of the screen is when the A pump is
when the A pump is pumping. Lower pressure at the beginning, middle or end of the chromatogram can
indicate which pump has lost prime.

Code 4: Unacceptable Total Pressure during Run (bar)
- Unacceptable minimum system pressure during analysis - Unacceptable minimum-maximum pressure
difference between pumps A & B during analysis
a. Check reagent level, confirm that bottles have enough reagent and tubing is reaching the reagent levels.
b. Check debubblers, make sure the air is bled out of them. Remember to check for air in debubblers with
every change of reagent A or B. c. From Main Menu, go to Devices → Pumps → Purge Pump. Turn
that the A and B reagent supply lines are primed completely. d. Remove the column and install
bypass union. From Devices → Pumps → Solvent Control, run both A and B
pumps at 50% each, 2ml/min, with reagents A and B selected (not wash) for 5 minutes. e. Remove
bypass union, install analytical column and activate the system. f. Run a Baseline and confirm that there is
no significant pressure drop within the run. NOTE: The first 1/3 of the screen is when the A pump is
when the A pump is pumping. Lower pressure at the beginning, middle or end of the chromatogram can
indicate which pump has lost prime. 2. Leak in System:
a. Check tubing connections for leaks and tighten. 3. System pressure too high: plugged 2μm frit or
column
a. Change 2μm frit with every column change.

Code 5: Unacceptable First Peak Retention Time (minutes)
- The non-glycated peak did not elute in the acceptable peak window
every change of reagent A or B. d. From Main Menu, go to Devices → Pumps → Purge Pump. Turn
tightened properly

Code 6: Unacceptable Second Peak Retention Time (minutes)
- The glycated peak did not elute in the acceptable peak window
tightened properly
Code 7: Unacceptable Control I Result
- Control level I material is not within the range stated on the package insert.
Note: If the chromatography is abnormal, refer to section 9.4 before performing the following steps.
1. Calibrate system. See Operator’s Manual section 6.3.2. 2. Incorrect dilution. Prepare new control
samples with the correct dilution ratio listed on the pack insert. 3. Reagent debubblers full of air. Bleed
the air out of the debubblers. 4. Control material. For lyophilized QC material, reconstituted QC material
and haemolysate dilutions of reconstituted QC material, ensure the correct storage, reconstitution and
expiration date, according to package insert(s).

Code 8: Unacceptable Control II Result
- Control level II material is not within the range stated on the package insert.
Refer to Code 7: Unacceptable Control I Result for appropriate Code 8 troubleshooting steps.
Code 9: Not Applicable for Calibration Run
Code 10: Not Applicable for Calibration Run
Code 11: Not applicable for Calibration Run
Code 12: Unacceptable Result Difference During Calibration (%)
- Replicate calibration injections demonstrate a % area difference of greater than 5%.
1. If the chromatography is abnormal, refer to section 9.4 before performing the following steps. 2. This
code indicates there is more than 5% mathematical difference in the result of the replicate calibrator
injections. This could indicate reproducibility issues during the system calibration. 3. Loss of Prime:
Air in the HPLC system.
a. Check physical reagent levels, confirm that bottles have enough reagent.

b. Check for leaks. c. Check debubblers, make sure the air is bled out of them. Remember to check for air
in debubblers with
is no significant pressure drop within the run. 4. Leak in System:
a. Check tubing connections for leaks and tighten.

Code 13: Calibration Slope Value
- Calibration slope greater than 2.0
1. Incorrect dilution. Prepare new calibration samples with the correct dilution ratio listed on the pack
insert. 2. Calibration material. For lyophilized QC material, reconstituted QC material and hemolysate
dilutions of reconstituted QC material, ensure the correct storage, reconstitution and expiration date,
according to package insert(s).

9.4 General Chromatography Troubleshooting
9.4.1 Normal Chromatography
Figure 9: Normal Chromatography Explained
The following explanations will give further detail of what to look for in the chromatography:
1. Baseline: Indicated by the green trace, this should be flat, however not completely flat. There will
always be some noise on the baseline when the pumps switch from A to B and back to A, as is evident
from the bumps present. This green trace displays the system noise being cancelled out from the final
result, and should not cause any alarm.
2. Pressure: It is paramount that the pressure remains steady throughout the run in order to achieve
optimal analysis conditions. Pressure is measured in units of Bar. Pressure fluctuations typically do not
exceed 1- 2 bar when system pressure is 10-25 Bar, and typically do not exceed 2-3 Bar when system
pressure is >25 bar. Within-run Δp of less than 20% will not be flagged with a Code 3 Peak Analysis &
Results Code and in the case of the Premier should be considered normal as long as QC values are within
range.
3. Peak Shape: The peaks should be sharp and resolute. The top of the first peak should not be much
wider than the second hash mark in the chromatography window. The second peak should return to
baseline and should not shoulder in any way, which would indicate full and efficient elution.

4. Valley: The valley should always return to baseline. Sometimes it is difficult for elevated HbA1c
samples to achieve a full return to the baseline, but it should be very close. Elevated baselines are
indicators of columns with high injection counts.
5. Tail: The second peak should return to baseline and should have plenty of time to allow the system to
re- equilibrate and elute all biological material off of the column before the next injection so as to avoid
carryover.
6. Retention Times: The retention times should remain consistent for each column. There may be minor
differences from column-to-column, but these will be ≈1% fluctuations at most.
7. Area counts: The system area count default settings will provide results from 250,000-2M. Ideal total
peak area counts are typically from 800,000-1,000,000.
8. Digital Pressure Monitor: This is the digital system pressure. For ideal pressure situations, see number
2
above.
9.4.2 General Recommendations
There are a number of key system factors that can affect chromatography, the below items are general
guidelines to aid in achieving the best system performance:
1. Ensure preventative maintenance activities are up to date.
2. Ensuring the activation settings are programmed as per user requirements will protect the system from
unnecessary activation/deactivation cycles.
3. It is recommended that the Premier Hb9210TM be deactivated daily in order to help eliminate
impurities
and salt build-up in the system.
4. Shorter standby times will aid in maintaining system performance by avoiding holding the analytical
column at 45oC in the column oven. Where possible, try to batch samples for analysis together.
5. As when storing columns before use, if the column will go unused for a period greater than 7 days it is
recommended to remove and store the column at 2-8oC.
6. The introduction of air into the system can cause the resin to dry out. In order to avoid this, ensure
debubblers are primed following changing of reagents.
9.4.3 Hb Variants & Other Potentially Interfering Substances
The presence of haemoglobin variants and other potentially interfering substances can impact upon some
HbA1c tests. Details of the robustness of the Premier Hb9210TM in the presence of common variants and
other potentially interfering substances are discussed in chapter 3 of this manual.

9.4.4 Air in the Pumps
Note the fluctuating pressure in the charted pressure reading (blue line). Possible Peak Analysis code
numbers: 2, 3, 5, 6, 7 & 8.
Figure 10: Chromatography with Air Bubble in B Pump
9.4.5 Short sample
Possible Peak Analysis code numbers: 1, 6, 7, 8 & 9.
Figure 11: Short Sample

9.5 Miscellaneous Operational Suggestions
The below points are not measures of periodic maintenance; they are precautionary measures that can be
taken, as needed.
9.5.1 Proper Handling of Biohazardous Waste
The Trinity Biotech Premier Hb9210TM boronate affinity glycated haemoglobin analyzer utilizes
reagents with 5% alcohol and analyzes very small volumes of blood samples. As such, it is not likely that
any infectious agents could reside in the liquid waste of the system. However, because of the possible
ecological risks of the instrument, operators must be trained in bio-hazardous waste training in order to
effectively reduce risk of contamination
Because of the ecological risks of medical devices, the decontamination and disposal procedures
associated with the instrument should be handled in accordance with the local regulations.
General recommendations are as follows:
✓ Appropriate decontamination is carried out if hazardous material comes in contact with the instrument.
✓ No hazardous decontamination or cleaning agents should be used as a result of a reaction with parts of
the
equipment or with material contained on it.
✓ Trinity Biotech or the applicable distribution partner should be immediately consulted if there is any
doubt about the compatibility of decontamination or cleaning agents with parts of the equipment or with
material contained on it.
When dumping the liquid waste, the use of universal protective precautions should be taken, including the
use of:
1. Protective goggles
2. Lab coat
3. Disposable gloves
The waste liquid should be dumped down a sink that goes to a sanitary sewer system. It should be flushed
with cold water so as to fully dilute. The waste tubing from the instrument should be wiped down with a
freshly prepared 10% bleach solution to decontaminate them while changing the waste container.
Household bleach is typically a 5.25% Sodium Hypochlorite solution. Spills on the external surfaces (of
the waste container or sink) should be wiped down with this bleach solution. After removal of safety gear,
hands should be washed thoroughly with soap and water.

9.6 Contacting Technical Support
If the troubleshooting steps outlined in this chapter fail to resolve an issue, please contact your technical
support team immediately. When consulting with service representatives, provide as much information as
possible in order to facilitate the most efficient and effective support possible.
Below is a non-exhaustive list of key information to provide when requesting support:
• Instrument Details
- Serial number - Module serial number (if applicable) - Instrument software version
• Description of the Issue
- A description of the issue being observed - Note of all error codes observed - Details of any
troubleshooting activities carried out to date - Preventative maintenance activities - Typical daily
instrument workload - Column injection count
• Chromatography
- Example chromatography that clearly shows the issue described - Lot number information for all
columns, reagents and consumables

PREMIER Hb9210TM Data
Transmission
CHAPTER CONTENTS
✓ ASTM E1394-97
Protocol ✓ Data Transmission using
Data Dump

Chapter 10 - Data Transmission/LIS Protocol
10.1 Premier Hb9210TM Data Transmission using ASTM E1394-97 Protocol
10.1.1 Introduction
Trinity Biotech’s HbA1c analyzer, the Premier Hb9210TM, has the ability to transmit data to a
Laboratory Information System, (LIS). There are two modes of transmitting data to an LIS, ASCII data
dump out of a “dumb” communication port and one that uses the ASTM E1394-97 and ASTM E1381-95
protocols.
10.1.2 General
Trinity Biotech’s Premier Hb9210TM HbA1c software is written specifically for HbA1c analysis on
Trinity Biotech’s analyzer, the Premier Hb9210TM. The laboratory runs any number of whole blood and
hemolysate samples in a batch mode. After analysis of the samples in the batch, all sample identifications
and results are sent to a host computer or a LIS in a batch mode, immediately after each sample, or both.
Note: all control data is transmitted, but calibrator data is not.
During analysis of the batch of samples, each sample will yield a chromatogram, or graph, of the
separated glycated and non-glycated hemoglobin proteins. The percentage area of the glycated
hemoglobin protein peak is calculated and results of %HbA1c and millimoles HbA1c/mol Hb (mM A1c)
are given. Laboratory (Operator) review of patient result chromatograms and QC recovery verification is
required for quality assurance as required under the Laboratory’s quality and GLP programs. The Premier
Hb9210TM system software is sufficiently sophisticated to facilitate this process by detection of common
“peak code” sample and system management errors and is designed to aid the operator in the detection
and prompt resolution of basic operational needs and adjustments. If peak codes are received during the
run and cause the run to stop after x occurrences, LIS transmission will stop immediately. A dash symbol
(-) and no result will be transmitted to the LIS in the event of detected sample and system management
errors.
Communication between the HbA1c software and the LIS is bi-directional, as prescribed by the ASTM
E1394-97 protocol, meaning that acknowledgement and ready signals are transmitted between the HbA1c
software and the LIS. But communication is uni-directional as to general data transfer, meaning that
results are transmitted to the LIS and run lists, test orders, and patient information are not downloaded to
the HbA1c software from the LIS.
Each patient set of results sent to the LIS will have a Header Record, a Patient Record, an Order Record,
five (5) Result Records, and a Message Terminator Record.
10.1.3 Physical Connection & Communication
On the front panel of the CPU Module on the Premier Hb9210TM is an RS-232-C, male, 9-pin connector
for interfacing the analyzer with a LIS; generally com port 2. A null modem cable is used to connect to
this port to a LIS. Communication is user selectable baud rate, number of data bits, parity, and stop bits.
The default is 9600 baud rate, 8 data bits, no parity, 1 stop bit.
1 | Chapter 10 – Data Transmission Rev. 23

10.2 ASTM E1394-97 Field Usage
10.2.1 Header Record
The Header Record starts with the “H” character (ASCII 72) and is followed by the characters used for
the Field Delimiter, “|” (ASCII 124); Repeat Delimiter, “\” (ASCII 92); Component Delimiter, “^”
(ASCII 94) and the Escape Character, “&” (ASCII 38). This first series of 5 characters constitute the 1st
field of the Header Record and as all fields, is followed by the Field Delimiter, “|”. The rest of the Header
Record fields are explained below:
# Field Value or Use 1 Record type ID H|\^&
(Stands for Header record and consists of 5 characters) 2 Control ID Not used. 3 Access password Not
used. 4 Instrument
Identity
ID of instrument in the format: “PREMIER^XXXXXX” (Quotation marks are not sent.) Component 1,
PREMIER, is the instrument type; component 2, XXXXXX, is the instrument serial number. 5 Sender
name / ID Not used. 6 Sender address Not used. 7 Reserved Not used. 8 Sender telephone
#
Not used.
9 Sender
characteristics
Not used.
10 Receiver Identity ASTM RECVR 11 Receiver ID Not used. 12 Comment /
instructions
Not used.
13 Processing ID P
(stands for Production) 14 Specification
version number
E 1394-97
15 Time stamp for message generation
Current date and time in the format: YYYYMMDDHHMMSS
As prescribed in the ASTM E1394-97 document that describes the protocol and fields of this usage, the
fields are separated with a defined Field Delimiter, “|”; empty or Not used fields will only contain the
Field

Delimiter. The final field may or may not end with the Field Delimiter. The end of the Header Record
will be indicated with a Carriage Return character (ASCII 13). Any ASTM E1394-97 prescribed fields
following the Carriage Return character will not be used and therefore not sent.
10.2.2 Patient Record
The Patient Record is used with minimal information. In this situation, the Patient Record is used only to
signal that new patient results are forthcoming. The Patient Record fields are explained below:
# Field Value or Use 1 Record type ID P
(Stands for Patient type record) 2 Sequence number 1 through 7, then
repeats.
The fields are separated with a defined Field Delimiter, “|”. The final field may or may not end with the
Field Delimiter. The end of the Patient Record will be indicated with a Carriage Return character (ASCII
13). Any ASTM E1394-97 prescribed fields following the Carriage Return character will not be used and
therefore not sent.
10.2.3 Order Record
The Order Record is also used with minimal information, as the HbA1c software does not accept order or
assay commands from the LIS. The Order Record fields are explained below:
# Field Value or Use 1 Record type ID O
(Stands for Order type record) 2 Sequence number 1
(Only one Order per patient.) 3 LIS assigned Specimen ID
Not used. 4 Instrument assigned Specimen
ID field
User entered (may be by barcode scanner) identification or accession number of patient sample. If no ID
or sample name is entered a single space, (ASCII 32), will be the entry. 5 Universal Test ID and
Manufacturer's Code
^^^PREMIER HBA1C Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component
Delimiters are given for them. The last component of this field

is PREMIER HBA1C, which is the Manufacturer's Code. 6 Priority R for Routine samples and S for
Stat samples. 7 Requested / Ordered Priority Not used. 8
Specimen Collection Date &
Time
Not used.
9 Collection End Time Not used. 10 Collection Volume Not used. 11 Collector ID Not used. 12 Action
Code Not used. 13 Danger Code Not used. 14 Relevant Clinical Info Not used. 15 Date / Time Specimen
Received Not used. 16 Specimen Descriptor Not used. 17 Ordering Physician Not used. 18 Physician's
Tel # Not used. 19 User Field #1 Control Lot Number will be
transmitted if “Transmit Control Lot Number” checkbox is selected. 20 User Field #2 Not used. 21 Lab
Field #1 Not used. 22 Lab Field #2 Not used. 23 Date/Time Results Reported Not used. 24 Instrument
Charge (billing) Not used. 25 Instrument Section ID R^VVV
Component 1 is the rack number of the sample, a number from 1 to 4; and component 2 is the vial
position number of the sample, a number from 001 to 010. Controls will use 0000^000 for this field
value. 26 Report Type F
(Stands for Final.)
The fields are separated with a defined Field Delimiter, “|”; empty or Not used fields will only contain the
Field Delimiter. The final field may or may not end with the Field Delimiter. The end of the Order Record
will be indicated with a Carriage Return character (ASCII 13). Any ASTM E1394-97 prescribed fields

10.2.4 Result Number 1 Record ... Result Number 5 Record
The Result Records 1 through 5 are the actual results of the assay of each patient sample. The HbA1c
software provides 2 results for each assay.
The Result Record 1 fields are explained below (legacy reporting units of %GHb):
# Field Value or Use 1 Record type ID R
(Stands for Results type record) 2 Sequence number 1
(Test result 1 of 5.) 3 Universal Test ID and
Manufacturer's Code
^^^GHb Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component Delimiters are
given for them. The last component of this field is GHb, which is the Manufacturer's Code. 4 Data Value
--- 5 Units of data %
The percent character, (ASCII 37). 6 Reference Ranges Not used. 7
Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
(Stands for Final) 10 Date of Change in
Instrument Normative Values field
Not used.
11 Operator Identification
field
Not used.
12 Date/Time Test Started
field
Not used.
13 Date/Time Test
Completed
Date and time in the format: YYYYMMDDHHMMSS 14 Instrument
Identification
Not used.
Field Delimiter. The final field may or may not end with the Field Delimiter. The end of the Results
Record will be indicated with a Carriage Return character (ASCII 13). Any ASTM E1394-97 prescribed
fields following the Carriage Return character will not be used and therefore not sent.

The Result Record 2 fields are explained below:
Manufacturer's Code
^^^HbA1c Part 1 of the Universal Test ID are blank and thus 1 Component Delimiter are given.. The last
component of this field is HbA1c, which is the Manufacturer's Code. 4 Data Value Value of 0.0 through
100.0 or 0.00
through 100.00, with one or two fixed decimal places. 5 Units of data %
The percent character, (ASCII 37). 6 Reference Ranges Not
used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality Testing Not used. 9 Status F
(Stands for Final) 10 Date of Change in Instrument
Normative Values field
Not used.
11 Operator Identification field Not used. 12 Date/Time Test Started field Not used. 13 Date/Time Test
Completed Date and time in the format:
YYYYMMDDHHMMSS 14 Instrument Identification Not
used.
Field Delimiter. The end of the Results Record will be indicated with a Carriage Return character (ASCII
therefore not sent.

The Result Record 3 fields are explained below (legacy reporting units of AG):
Manufacturer's Code
^^ÂG Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component Delimiters are
given for them. The last component of this field is AG, which is the Manufacturer's Code. 4 Data Value
--- 5 Units of data mg/dl 6 Reference Ranges Not used. 7 Result Abnormal Flags Not used. 8 Nature of
Abnormality Testing Not used. 9 Status F
Not used.
used.

Manufacturer's Code
^^^mMA1c Part 3 of the Universal Test ID is blank and thus a Component Delimiter is given for that
place. Components of this field is mM A1c, which is the Manufacturer's Code. 4 Data Value Value of 0.0
through 999.9, with one
fixed decimal place or no decimal places. 5 Units of data mMolHbA1c/MolHb 6 Reference Ranges Not
used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
YYYYMMDDHHMMSS 14 Instrument Identification Not used.
The fields are separated with the Primus defined Field Delimiter, “|”; empty or Not used fields will only
contain the Field Delimiter. The end of the Results Record will be indicated with a Carriage Return
character (ASCII 13). Any ASTM E1394-97 prescribed fields following the Carriage Return character
will not be used and therefore not sent.

Manufacturer's Code
^^^Code Part 3 of the Universal Test ID is blank and thus a Component Delimiter is given for that place.
Components of this field is Code, which is the Manufacturer's Code. 4 Data Value Value of 1 through 11,
more than one
code could be displayed. 5 Units of data Not used. 6 Reference
Ranges Not used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
therefore not sent.

10.2.5 Message Terminator
The Message Terminator is sent once all the results have been sent to the LIS. The Message Terminator
fields are explained below:
# Field Value or Use 1 Record type ID L
(Stands for Message Terminator record) 2 Sequence number 1
Field Delimiter. The end of the Message Terminator Record will be indicated with a Carriage Return
10.2.6 Communication to LIS Example
The following is an example of communication to a Laboratory Information System (LIS) and the
computer on the Premier Hb9210TM analyzing hemoglobin A1c samples using the software.
10.2.7 Bi-directional Communication between the Premier Hb9210TM and the LIS
computer on the Premier Hb9210TM analyzing glycated hemoglobin samples using the software from
system 100051.
RX: H|\^&|||PREMIER^100051||||||ASTM RECVR|||P|
E 1394-97|20090519153949<0D> RX: P|1<0D> RX: O|1||Sam Smith|^^^PREMIER
HBA1C|R|||||||||||||||||||WX61^001|F<0D> RX: R|1|^^^GHb|---|%||||F||||20090519153950||<0D> RX:
R|2|^^^HbA1c|9.0|%||||F||||20090519153950|||<0D> RX:
R|3|^^ÂG|---|mg/dl||||F||||20090519153950|||<0D> RX:
R|4|^^^mMA1c|73.2|mMolHbA1c/MolHb||||F||||20090519153950|||<0D> RX: R|5|^^^Code|1 8 12
|||||F||||20090519153950||<0D>
RX: P|2<0D> ...... ...... RX: L|1<0D>
Where: RX is Premier transmitted records (received by LIS)
<0D> is a Carriage Return

10.3 Premier Hb9210TM Data Transmission using Data Dump
10.3.1 Introduction
Trinity Biotech’s HbA1c analyzer, the Premier Hb9210TM, has the ability to transmit data to a
Laboratory Information System, (LIS). There are two modes of transmitting data to an LIS, ASCII data
dump out of a “dumb” communication port and one that uses the ASTM E1394-97 and ASTM E1381-95
protocols.
10.3.2 General
Trinity Biotech’s Premier Hb9210TM HbA1c software is written specifically for HbA1c analysis on
Trinity Biotech’s analyzer, the Premier Hb9210TM.
The laboratory runs any number of whole blood and haemolysate samples in a batch mode. After analysis
of the samples in the batch, the sample identifications and results are sent to a host computer or a LIS in a
batch mode, immediately after each sample, or both. During analysis of the batch of samples, each sample
will yield a chromatogram, or graph, of the separated glycated and non-glycated hemoglobin proteins.
The percentage area of the glycated hemoglobin protein peak is calculated and results of %HbA1c and
millimoles HbA1c/mol Hb (mM A1c) are given. Please note that analysis is transmitted to a LIS, each
chromatogram should have a trained user quickly look at each chromatogram for acceptability. The
results are then transmitted to the LIS.
Communication between the HbA1c software and the LIS is uni-directional. This option of data transfer
does not use any acknowledgment of data sent; that is to say it is a “dumb” data dump.
10.3.3 Physical Connection & Communication
On the front panel of the CPU Module on the Premier Hb9210TM is an RS-232-C, male, 9-pin pin
connector for interfacing the analyzer with a LIS; generally com port 2. A null modem cable is used to
connect to this port to a LIS. Communication is user selectable baud rate, number of data bits, parity, and
stop bits. The default is 9600 baud rate, 8 data bits, no parity, 1 stop bit.
10.3.4 Data Field Usage
A record will start with “/TRINITY BIOTECH PREMIER”. Then identified by what type of result record,
“(Individual)” or “(Batch)” and followed with the Field Delimiter, “|” (ASCII 124). Following is the name
of the system, batch number, rack position, and sample ID. Then the results are given with the proper
units. Each of the fields are separated by the Field Delimiter, “|” (ASCII 124). The record ends with a
carriage return and a line feed (ASCII 13 and ASCII 10).
Each record will be treated as having a varying number of fields with the following conditions...
✓ Each field is delimited with a “|” character.

✓ The first field will start with a “/TRINITY BIOTECH PREMIER”. Then identified by what type of
result record,
“(Individual)” or “(Batch)”. The first field will be consistent with each version of software.
✓ Subsequent fields may be in any order. These fields will have the general form:
“Fieldname=Value&Units”
1. System Serial#= 2. Batch= 3. Position=Rack:Vial 4. ID= 5. GHb=--- 6. HbA1c=xx.x% or xx.xx% 7.
AG=--- 8. mMA1c=xxx mMolHbA1c/MolHb or xxx.x mMolHbA1c/MolHb 9. Code=xx (if enabled) 10.
Data Points=x,x,x,x,x... (if enabled)
10.3.5 Premier Hb9210TM Communication to LIS Example
computer on the Premier Hb9210TM analyzing glycohaemoglobin samples using the software from
system 100051, batch 3129.
10.3.6 Uni-directional Communication Between Premier Hb9210TM and the LIS
RX: /TRINITY BIOTECH PREMIER (Batch)|System Serial#=100051 |Batch=3129|
Position=WX6I:001|ID=Sam Smith |GHb=---|HbA1c=8.9%| AG=---|mMA1c=72.5
mMolHbA1c/MolHb|Code=3 4 10|Data Points=1,1,-1,-3,-2,-1,-
1,0,1,39,442,3113,9440,13952,12531,8385,4742,2455,1209,632,379,257,194,158,133,116,102,9
2,83,77,71,65,64,64,62,65,111,295,446,427,351,294,261,243,233,222,206,184,157,131,109,87,6
9,54,44,37,34,33,30,26 <0D><0A>
Where: RX is Premier transmitted records (received by LIS)
<0D> is a Carriage Return <0A> is a Line Feed


PREMIER Hb9210TM TLA Appendix
CHAPTER CONTENTS
✓ TLA Premier Connection ✓ TLA Premier Operation ✓ TLA LIS Protocol

TLA Appendix – TLA Premier Hb9210TM
TLA 1. System Overview
This section provides a physical overview of the TLA Premier Hb9210TM.
Physical TLA Connection Overview
The TLA Premier Hb9210TM is connected to the Inpeco track system via an Interface Module. The
Interface Module transports sample tubes towards the analyser, whereby the analyser can aspirate the
sample via the Probe Needle. During sampling, the sample tube is held in place by the Tube Locker. Once
the sampling has completed, the Tube Locker releases the sample and it is routed back onto the main
track system.
Figure 1 below is a plan view of the main track, outlined in orange, connected to the Interface Module,
outlined in green. The TLA Premier Hb9210TM sits on top of the Interface Module and is outlined in a
dashed blue line. A reference to each important stage in the sampling process is listed below Figure 1.
Figure 1: Plan View - TLA Premier Hb9210TM Interface Module & Main Track
1 | TLA Appendix Rev. 22

1. The location of the TLA Premier Hb9210TM Analyser.
2. The location where the sample tube is diverted from the main lane to the pit lane of the TLA Premier
Hb9210TM .
3. The location where the sample tube leaves the main track and enters the inward belt on the Interface
Module.
4. The location were the sample tubes stop and wait while there is a current sample tube under the Probe.
5. The transport grip that moves the sample tube towards and away from the Probe.
6. The location where the sample tube is under the Probe during sampling.
7. The location where the sample tube enters the outward belt on the Interface Module.
8. The location where the sample tube returns back onto the main track.
Figure 2 below highlights the items the Operator will interact with on TLA Interface Module, namely the
External and Internal TLA Buttons and the Carousel Wheel. Please note that the Premier Hb9210TM
instrument has been removed from this image to give a clearer view of the TLA Interface Module.
Figure 2: TLA Buttons & Carousel Wheel
1. External TLA Button – depending on system status can light up green, red, yellow or white.
2. Internal TLA Button – lights up blue when active.
3. Carousel Wheel – the wheel has 10 positions, first 4 positions are used for calibrators and controls, the
remaining 6 positions are used for Priority Samples.
3
1
2

TLA 2. System Operation
The sections below provide instructions on how to enable TLA connection, how to calibrate, run samples
and set the Control Scheme on the TLA Premier Hb9210TM.
Enable TLA System Connection
✓ From the Main Menu, select Edit → System
Parameters icon (see Figure 3).
Figure 3: System Parameters
✓ In the System Parameters window, ensure that ‘Enable connection of Inpeco TLA System’ is checked
(see Figure 4).
Figure 4: System Parameters Window
✓ LIS Communication parameters should be chosen based upon individual customer requirements (see
Figure 5).
Figure 5: LIS Communication

TLA System Calibration
Prior to running a system calibration, the system must have completed steps 1 & 2:
1. Activation – see section 6.3.1
2. Baseline – see section 6.3.3
Then continue with the following steps:
3. Return to the Main Menu by touching or clicking the ‘Show User Interface’ button on the top left of the
main Premier Hb9210TM assay screen.
4. From the Main Menu screen, select the ‘Operation’ icon.
5. From the Operation Menu, select the ‘Run Calibration’ option shown below. This process includes a
baseline noise calibration to cancel out any system noise (e.g. Pumps, other vibrations) from the final
chromatography.
Figure 6: Operation Menu - Run Calibration Selection
The Calibration Run dialogue shown in Figure 7. NOTE: The 1x option has been disabled. Only the 2x
and 3x options are available (for improved calibration accuracy).
6. By selecting the Run Controls Only option, the system will simply run Control I, then Control II (and
not the calibrators).
7. Select the Run button, circled in Red in Figure 7. This will initiate the Calibration Run but the run will
not begin until the Calibrators and controls are placed on the TLA Carousel Wheel and the External TLA
button is pressed.
Figure 7: Run Calibration Confirmation Dialogue

8. Press the External TLA button (Figure 2 Item 1). The button should turn solid red. This allows access
to the
Carousel Wheel.
9. The Internal TLA button located beside the Carousel Wheel will now light up solid blue (Figure 2 Item
2). Open
the Carousel Access Cover and load the Calibrators and Controls on the Carousel Wheel (Figure 2 Item
3).
10. Pressing the Internal TLA button will rotate the TLA Carousel Wheel one position at a time. The
Calibrators and
Controls should be placed on the wheel in the following order and locations:
✓ Position 1: Calibrator 1
✓ Position 2: Calibrator 2
✓ Position 3: Control I
✓ Position 4: Control II
Figure 8: Calibrator and Control Locations
11. Once the Calibrators and Controls are loaded, close the Carousel Access Cover.
12. Press the External TLA button. Once pressed, the button will flash yellow. It will then turn solid
yellow and the
Calibration will begin.
When the Carousel Access Cover is opened, Position 1 will be first position presented. There are 10 positions in
total. The first 4 positions are for the Calibrators and Controls. The remaining 6 positions are for Priority Samples.
Pressing the Internal TLA button will move to the next position. When position 10 is reached, pressing the button
one more time will revert the wheel back to position 1.

Running Priority Samples
Prior to running Priority Samples, the system must be Activated and Calibrated.
1. From the Main Menu, select Operation then
select Priority Samples (see Figure 9).
Figure 9: Priority Samples
2. In the Priority Samples menu, check the required priority samples to run and press the Run icon. Figure
10
illustrates 3 priority samples to be run in positions 5, 6 and 7.
Figure 10: Priority Sample Run Menu
There are 6 positions for Priority Samples. They are positions 5, 6, 7, 8, 9 and 10 on the Carousel Wheel. If the
system is currently processing samples from the track, and the Priority Sample Run button is pressed in the SW, the
system will finish the current sample and then go to stand by. It will then process all priority samples. All remaining
samples from the track will then be processed after the Priority Samples.

to the
Carousel Wheel (Figure 2 Item 3).
2). Open
the Carousel Access Cover and load the number Priority Samples that were selected in Figure 10.
5. Once the Priority Samples are loaded, close the Carousel Access Cover.
6. Press the External TLA button. Once pressed, the button will flash yellow. It will then turn solid yellow
and the
Priority Sample run will begin.
Accessing the TLA Carousel Wheel
The TLA Carousel Wheel can be accessed when the system is not running, to remove sample tubes, by
following the steps below:
to the
Carousel Wheel (Figure 2 Item 3).
2). Open
the Carousel Access Cover and remove the desired tubes.
Control Scheme
The Control Scheme can be customised to run Controls at specific intervals throughout a batch run. The
instructions below detail how to set the Control Scheme:
1. From the Main Menu, select Edit then select
Control Scheme icon (see Figure 11).
Figure 11: Control Scheme Icon
2. In the Control Scheme window, it is possible to:
✓ Alternate Control I & II every interval – selecting this option and setting the interval to 0, no controls
will run. By setting a desired interval number, Control I will run after the desired number of samples
followed by Control II at the next interval of samples. This pattern will repeat until the batch run
completes. See Figure 12.
✓ Run Control I & II every interval – selecting this option and setting the interval to 0, no controls will
run. By setting a desired interval number, Control I & II will run after the desired number of samples.
This pattern will repeat until the batch run completes. See Figure 12.

Figure 12: Control Scheme Options

TLA 3. TLA LIS Protocol
LIS Transmission (ASTM E1394)
Each patient set of results sent to the LIS will have:
✓ Header Record – only sent once at beginning of transmission. ✓ Patient Record ✓ Order Record ✓
Six Result Records ✓ Message Terminator Record – only sent once at end of transmission.
Header Record
The Header Record starts with the “H” character (ASCII 72) and is followed by the characters used for
the Field Delimiter, “|” (ASCII 124); Repeat Delimiter, “\” (ASCII 92); Component Delimiter, “^”
(ASCII 94) and the Escape Character, “&” (ASCII 38). This first series of 5 characters constitute the 1st
field of the Header Record and as all fields, is followed by the Field Delimiter, “|”. The rest of the Header
Record fields are explained below:
# Field Value or Use 1 Record type ID H|\^&
(Stands for Header record and consists of 5 characters) 2 Control ID Not used. 3 Access password Not
used. 4 Instrument
Identity
ID of instrument in the format: “PREMIER^XXXXXX” (Quotation marks are not sent.) Component 1,
PREMIER, is the instrument type; component 2, XXXXXX, is the instrument serial number. 5 Sender
name / ID Not used. 6 Sender address Not used. 7 Reserved Not used. 8 Sender telephone
#
Not used.
9 Sender
characteristics
Not used.
10 Receiver Identity ASTM RECVR 11 Receiver ID Not used. 12 Comment /
instructions
Not used.
13 Processing ID P
(stands for Production)

14 Specification
version number
E 1394-97
15 Time stamp for message generation
Current date and time in the format: YYYYMMDDHHMMSS
As prescribed in the ASTM E1394-97 document that describes the protocol and fields of this usage, the
fields are separated with a defined Field Delimiter, “|”; empty or Not used fields will only contain the
Field Delimiter. The final field may or may not end with the Field Delimiter. The end of the Header
Patient Record
The Patient Record is used with minimal information. In this situation, the Patient Record is used only to
signal that new patient results are forthcoming. The Patient Record fields are explained below:
# Field Value or Use 1 Record type ID P
(Stands for Patient type record) 2 Sequence number 1 through 7, then
repeats.
Field Delimiter. The end of the Patient Record will be indicated with a Carriage Return character (ASCII
therefore not sent.
Order Record
The Order Record is also used with minimal information, as the HbA1c software does not accept order or
assay commands from the LIS. The Order Record fields are explained below:
# Field Value or Use 1 Record type ID O
(Stands for Order type record) 2 Sequence number 1
(Only one Order per patient.) 3 LIS assigned Specimen ID
Not used. 4 Instrument assigned Specimen
ID field
User entered (may be by barcode scanner) identification or accession number of patient sample. If no ID
or sample

name is entered a single space, (ASCII 32), will be the entry. 5 Universal Test ID and
Manufacturer's Code
^^^PREMIER HBA1C Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component
Delimiters are given for them. The last component of this field is PREMIER HBA1C, which is the
Manufacturer's Code. 6 Priority R for Routine samples and S for
Stat samples. 7 Requested / Ordered Priority Not used. 8
Specimen Collection Date &
Time
Not used.
9 Collection End Time Not used. 10 Collection Volume Not used. 11 Collector ID Not used. 12 Action
Code Q for Control samples otherwise
empty 13 Danger Code Not used. 14 Relevant Clinical Info
Not used. 15 Date / Time Specimen Received Not used. 16 Specimen Descriptor Blood type. H for
haemolysate
samples, A for anaemic samples and empty for whole blood samples. 17 Ordering Physician Not used. 18
Physician's Tel # Not used. 19 User Field #1 Not used. 20 User Field #2 Not used. 21 Lab Field #1 Not
used. 22 Lab Field #2 Not used. 23 Date/Time Results Reported Not used. 24 Instrument Charge (billing)
Not used. 25 Instrument Section ID TLA^000
TLA^001 for STAT samples only 26 Report Type F
(Stands for Final.)
Field Delimiter. The final field may or may not end with the Field Delimiter. The end of the Order Record
will

be indicated with a Carriage Return character (ASCII 13). Any ASTM E1394-97 prescribed fields
The Premier HbA1c software provides 4 results for each assay. The Result Records 1 through 4 are the
actual results of the assay of each patient sample.
Manufacturer's Code
^^^GHb Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component Delimiters are
given for them. The last component of this field is GHb, which is the Manufacturer's Code. 4 Data Value
--- 5 Units of data %
The percent character, (ASCII 37). 6 Reference Ranges Not used. 7
Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
11 Operator Identification
field
Not used.
12 Date/Time Test Started
field
Not used.
13 Date/Time Test
Completed
Date and time in the format: YYYYMMDDHHMMSS 14 Instrument
Identification
Not used.

Manufacturer's Code
^^^HbA1c Part 1 of the Universal Test ID are blank and thus 1 Component Delimiter are given. The last
component of this field is HbA1c, which is the Manufacturer's Code. 4 Data Value Value of 0.0 through
100.0 or 0.00
through 100.00, with one or two fixed decimal places. 5 Units of data %
The percent character, (ASCII 37). 6 Reference Ranges Not
used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality Testing Not used. 9 Status F
Not used.
used.
therefore not sent.
The Result Record 3 fields are explained below (legacy reporting units of AG):
(Stands for Results type record)

2 Sequence number 3
Manufacturer's Code
^^ÂG Parts 1 through 3 of the Universal Test ID are blank and thus only 3 Component Delimiters are
given for them. The last component of this field is AG, which is the Manufacturer's Code. 4 Data Value
--- 5 Units of data mg/dl or mmol/l
User selectable. For milligrams per deciliter or millimoles per litre. 6 Reference Ranges Not used. 7
Result Abnormal Flags Not used. 8 Nature of Abnormality Testing Not used. 9 Status F
Not used.
used.

Manufacturer's Code
^^^mMA1c Part 3 of the Universal Test ID is blank and thus a Component Delimiter is given for that
place. Components of this field is mMA1c, which is the Manufacturer's Code. 4 Data Value Value of 0.0
through 999.9, with one
fixed decimal place or no decimal places. 5 Units of data mMolHbA1c/MolHb 6 Reference Ranges Not
used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
The fields are separated with the Primus defined Field Delimiter, “|”; empty or Not used fields will only
contain the Field Delimiter. The end of the Results Record will be indicated with a Carriage Return

The Result Record 5 fields are explained below (if transmitting codes is disabled, this record will be
deleted):
Manufacturer's Code
^^^Code Part 3 of the Universal Test ID is blank and thus a Component Delimiter is given for that place.
Components of this field is Code, which is the Manufacturer's Code. 4 Data Value Value of 1 through 12,
more than one
code could be displayed. 5 Units of data Not used. 6 Reference
Ranges Not used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
therefore not sent.

The Result Record 6 fields are explained below (if chromatogram points is disabled, this record will be
deleted):
Manufacturer's Code
^^^Data Points Part 3 of the Universal Test ID is blank and thus a Component Delimiter is given for that
place. Components of this field is Data Points, which is the Manufacturer's Code. 4 Data Value Value of
integers. Total of 60 averaged data points (sampled from all the data points) are displayed. 5 Units of data
Not used. 6 Reference Ranges Not used. 7 Result Abnormal Flags Not used. 8 Nature of Abnormality
Testing
Not used.
9 Status F
Not used.
therefore not sent.

Message Terminator Record
The Message Terminator is sent once all the results have been sent to the LIS. The Message Terminator
fields are explained below:
# Field Value or Use 1 Record type ID L
(Stands for Message Terminator record) 2 Sequence number 1
Field Delimiter. The end of the Message Terminator Record will be indicated with a Carriage Return
Example of ASTM E1394 Transmission between Premier and the LIS
Records Received by LIS:
RX: H|\^&|||PREMIER^XXXXXX||||||ASTM RECVR|||P| E 1394-97|20090519153949<0D> RX:
P|1<0D> RX: O|1||Jon Davis|^^^PREMIER HBA1C|R|||||||||||||||||||TLA^000|F<0D> RX:
R|1|^^^GHb|---|%||||F||||20090519153950||<0D> RX: R|2|^^^HbA1c|9.0|%||||F||||20090519153950||<0D>
RX: R|3|^^ÂG|---|mg/dl||||F||||20090519153950||<0D> RX:
R|4|^^^mMA1c|73.2|mMolHbA1c/MolHb||||F||||20090519153950||<0D> RX: R|5|^^^Code|1 8 12
|||||F||||20090519153950||<0D> (if transmitting codes is disabled, this record will be deleted) RX:
R|6|^^^Data Points|1,1,-1,-3,-2,-1,-
1,0,1,39,442,3113,9440,13952,12531,8385,4742,2455,1209,632,379,257,194,158,133,
116,102,92,83,77,71,65,64,64,62,65,111,295,446,427,351,294,261,243,233,222,206,18
4,157,131,109,87,69,54,44,37,34,33,30,26 |||||F||||20090519153950||<0D> (if transmitting chromatogram
points is disabled, this record will be deleted) RX: P|2<0D> ...... ...... RX: L|1<0D> Where: RX is Premier
transmitted records (received by LIS) <0D> is a Carriage Return

Host Query Message Format
1. Query message
This section describes the query message structure and each record.
Query message structure (Premier Instrument -> PC (LIS))
Query header record H Query Record Q Message End Record L
Query record is composed of 13 fields.
# Field Description Remark (Example) 1 Record Type Set Character “Q” 2 Sequence No. Set Digit “1” 3
Patient ID Sample ID (*1,*2,*3) “123456789012345” 13 Query Code Indicates the content of query
(Request an order)
“O”
End of record End of record <CR> (0x)D)
Example 1: Q|1|123456789012345||||||||||O<CR>
Example 2: Q|1|12345678901234567890123||||||||||O<CR>
(*1) Sample ID consists of one component only. (*2) Sample ID has no length restriction. (*3) Sample ID
is not padded with any character. The ‘|’ character should be used for splitting.
2. Response Message
Response message structure (PC (LIS) -> Premier Instrument)
Message header record H Patient information record P Test order record O Message end record L
✓ Patient information record is composed of 2 fields.
# Field Description Remark (Example) 1 Record Type Set Character “P”

2 Sequence No. Sequence number starting with “1” “1”
End of record End of record <CR> (0x0D) Example: P|1<CR>
✓ Test Order Record
Test order record is composed of 16 fields, which are different from those of the measurement result
message.
# Field Description Remark (Example) 1 Record Type Set Character “O” 2 Sequence No. Set digit “1” 3
Patient ID Sample ID (*1,*2,*3) “123456789012345” 5 Measurement
Item
Measurement item name (either of below)
1. Measure (*4) 2. No measure
1. “^^^HbA1c” 2. NULL (No output)
6 Priority Priority Code (Routine Sample) “R” 16 Sample type Sample type (either of below)
1. Whole blood Sample (default) 2. Haemolysate Sample 3. Anemic Sample
1. NULL 2. “H” 3. “A”
End of record End of record <CR> (0x0D)
Example 1: O|1|123456789012345||^^^HbA1c|R||||||||||H<CR>
Example 2: O|1|12345678901234567890123||^^^HbA1c|R||||||||||H<CR>
(*1) Sample ID consists of one component only. (*2) Sample ID has no length restriction. (*3) Sample ID
is not padded with any character. The ‘|’ character should be used for splitting. (*4) Measurement item
consists of four components but the first three are not used.


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Premier Hb9210TM Boronate Affinity | 09-00-0001 | 11-00-0001 | Revision 23

HAEMOGLOBINS SYSTEMS DIVISION

Chapter 2: Introduction & System Description

Chapter3: Principles of Operation

Chapter 5: Hardware Components

Chapter 7: Results & Interpretation

Chapter 9: Operator Troubleshooting

1.2 System Description

1.3 System Requirements

1.7 Important Safety Instructions

2.4 CPU Specifications

2.7 510(K) Listing

3.2 Principles of Analysis

3.3 Merits of the Assay

) 0.07 0.05 0.09 Repeatability %CV = (S

/mean)*100 1.26 0.72 0.85 Estimate of within-device precision SD, (S

) 0.09 0.09 0.16 Within-device precision %CV = (S

/mean)*100 1.62 1.28 1.50 Table 4: Intra-run Precision – Overview

) 0.07 0.06 0.09 Repeatability %CV = (S

/mean)*100 1.24 0.79 0.78 Estimate of within-device precision SD, (S

) 0.10 0.11 0.18 Within-device precision %CV = (S

/mean)*100 1.84 1.58 1.62

) 0.05 0.06 0.09 Repeatability %CV = (S

/mean)*100 0.85 0.79 0.85 Estimate of within-device precision SD, (S

) 0.08 0.09 0.18 Within-device precision %CV = (S

/mean)*100 1.47 1.20 1.62

) 0.10 0.04 0.10 Repeatability %CV = (S

/mean)*100 1.68 0.57 0.92 Estimate of within-device precision SD, (S

) 0.09 0.07 0.14 Within-device precision %CV = (S

/mean)*100 1.55 1.04 1.27 Table 5: Intra-run Precision - Individual Sites –

4.2 Reagent Use and Storage

4.4 Column Use and Storage

4.5 Specimen Requirements

4.6 Limitations of the Assay

5.3 Pump Module (09-30-2000S)

5.5 Detector Module (09-30-1700S)

5.8 Automatic Debubblers & Waste Pump

6.2 Installation Screens

6.5 Other Menus & Software Tools

6.6 Directory Structure

Chapter 7 – Results & Interpretation

7.5 Other Reports

8.3 Changing the Column

8.4 Detector Maintenance

8.5 Pump Maintenance

8.6 Sample Transport, Syringe Module and Probe Module Maintenance

Contact your Trinity Biotech Technical Representative:

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