SYNCHRON System(s) ApoB

Chemistry Information Sheet Apolipoprotein B

© 2020 Beckman Coulter, Inc. All rights reserved.
467905

For In Vitro Diagnostic Use

Rx Only

ANNUAL REVIEW
Reviewed by Date Reviewed by Date

PRINCIPLE
INTENDED USE

ApoB reagent, when used in conjunction with UniCel DxC 600/800 System(s) and SYNCHRON Systems APO Calibrator,
is intended for quantitative determination of apolipoprotein b concentration in human serum or plasma.

CLINICAL SIGNIFICANCE

Apolipoprotein B is the major protein constituent of low-density lipoprotein (LDL) and also constitutes about 40 percent
of the protein moiety of very-low-density lipoprotein (VLDL) and chylomicrons. Apolipoprotein B is responsible for lipid
transport and clearance. Measurement of apolipoprotein B aids in identifying coronary artery disease.1

METHODOLOGY

ApoB reagent is used to measure the apolipoprotein B concentration by turbidimetric method. In the reaction,
apolipoprotein B combines with specific antibody to form insoluble antigen-antibody complex.
The SYNCHRON System(s) automatically dilutes sample and proportions the appropriate sample and reagent volumes
into the cuvette. The ratio used is one part diluted sample to 11.5 parts reagent. The system monitors the change in
absorbance at 340 nanometers. This change in absorbance is directly proportional to the concentration of apolipoprotein
B in the sample and is used by the System to calculate and express apolipoprotein B concentration.

CHEMICAL REACTION SCHEME

In the performance of the ApoB Test, antibody to human apolipoprotein B is brought into contact with human
apolipoprotein B in a sample. The increase in absorbance (as light scatter) resulting from the antigen-antibody reaction
is measured and is a function of the sample apolipoprotein B concentration.2,3 Following calibration, the response for a
particular assay is automatically converted to concentration units by the analyzer.

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SPECIMEN
TYPE OF SPECIMEN

Biological fluid samples should be collected in the same manner routinely used for any laboratory test.4 Freshly drawn
serum or plasma are the preferred specimens. Acceptable anticoagulants are listed in the PROCEDURAL NOTES
section of this chemistry information sheet. Whole blood or urine are not recommended for use as a sample.

SPECIMEN STORAGE AND STABILITY

1. Tubes of blood are to be kept closed at all times and in a vertical position. It is recommended that the serum or
plasma be physically separated from contact with cells within two hours from the time of collection.5
2. Separated serum or plasma should not remain at room temperature longer than 8 hours. If assays are not
completed within 8 hours, serum or plasma should be stored at +2°C to +8°C. If assays are not completed within
48 hours, or the separated sample is to be stored beyond 48 hours, samples should be frozen at -15°C to -20°C.
Frozen samples should be thawed only once. Analyte deterioration may occur in samples that are repeatedly
frozen and thawed.5
Additional specimen storage and stability conditions as designated by this laboratory:

SAMPLE VOLUME

The optimum volume, when using a 0.5 mL sample cup, is 0.3 mL of sample. For optimum primary sample tube volumes
and minimum volumes, refer to the Primary Tube Sample Template for your system.

CRITERIA FOR UNACCEPTABLE SPECIMENS

Refer to the PROCEDURAL NOTES section of this chemistry information sheet for information on unacceptable
specimens.
Criteria for sample rejection as designated by this laboratory:

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PATIENT PREPARATION

Special instructions for patient preparation as designated by this laboratory:

SPECIMEN HANDLING

Special instructions for specimen handling as designated by this laboratory:

REAGENTS
CONTENTS

Each kit contains the following items:

Two ApoB Reagent Cartridges (2 x 100 tests)
One Lot-Specific Parameter Card

VOLUMES PER TEST

Sample Dilution Volumes

Sample Volume 15 µL
Diluent Volume 285 µL
Diluted Sample Volume (1:20 dilution) 20 µL
Total Reagent Volume 230 µL
Cartridge Volumes
A 200 µL
B – –
C 30 µL

REACTIVE INGREDIENTS

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 REAGENT CONSTITUENTS
Reaction Buffer 30 mL
Goat Antibody Monospecific for Human Apolipoprotein B 4.2 mL
Also non-reactive chemicals necessary for optimal system performance.

CAUTION
Sodium azide preservative may form explosive compounds in metal drain lines.
See NIOSH Bulletin: Explosive Azide Hazard (8/16/76).
To avoid the possible build-up of azide compounds, flush wastepipes with
water after the disposal of undiluted reagent. Sodium azide disposal must be in
accordance with appropriate local regulations.

GHS HAZARD CLASSIFICATION

Not classified as hazardous

Safety Data Sheet is available at techdocs.beckmancoulter.com

MATERIALS NEEDED BUT NOT SUPPLIED WITH REAGENT KIT

SYNCHRON Systems APO Calibrator

DIL 1 Cartridge
At least two levels of control material

REAGENT PREPARATION

Reagent can be removed from refrigerator and used immediately. Prior to placing reagent cartridge on system, mix
thoroughly by inversion.

ACCEPTABLE REAGENT PERFORMANCE

The acceptability of a reagent is determined by successful calibration and by ensuring that quality control results are
within your facility's acceptance criteria.

REAGENT STORAGE AND STABILITY

ApoB reagent when stored unopened at +2°C to +8°C will obtain the shelf-life indicated on the cartridge label. Once
opened, the reagent is stable for 30 days at +2°C to +8°C unless the expiration date is exceeded. DO NOT FREEZE.
DIL 1 stored unopened at room temperature is stable until the expiration date indicated on each cartridge. Once opened,
DIL 1 is stable for 60 days on instrument or until the expiration date, if sooner.

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Reagent storage location:

CALIBRATION
CALIBRATOR REQUIRED

SYNCHRON Systems APO Calibrator

CALIBRATOR PREPARATION

1. Allow calibrator to come to room temperature prior to use.

2. Mix thoroughly by inversion prior to use.

CALIBRATOR STORAGE AND STABILITY

The SYNCHRON Systems APO Calibrator is stable until the expiration date printed on the calibrator bottle if stored
capped at -15°C to -20°C. Thawed calibrators that are resealed and stored at +2°C to +8°C are stable for 90 days unless
the expiration date is exceeded.

CAUTION
Because this product is of human origin, it should be handled as though capable
of transmitting infectious diseases. Each serum or plasma donor unit used
in the preparation of this material was tested by United States Food and Drug
Administration (FDA) approved methods and found to be negative for antibodies
to HIV and HCV and nonreactive for HbsAg. Because no test method can offer
complete assurance that HIV, hepatitis B virus, and hepatitis C virus or other
infectious agents are absent, this material should be handled as though capable
of transmitting infectious diseases. This product may also contain other human
source material for which there is no approved test. The FDA recommends such
samples to be handled as specified in Centers for Disease Control's Biosafety
Level 2 guidelines.6

Calibrator storage location:

CALIBRATION INFORMATION

1. The system must have lot-specific parameters and a valid calibration adjustment in memory before controls or
patient samples can be run.
2. Under typical operating conditions the ApoB reagent cartridge must be calibrated every 14 days and also with
certain parts replacements or maintenance procedures, as defined in the UniCel DxC 600/800 System Instructions

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 For Use (IFU) manual. This assay has within-lot calibration available. Refer to the UniCel DxC 600/800 System
Instructions For Use (IFU) manual for information on this feature.
3. For detailed calibration instructions, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.
4. The system will automatically perform checks on the calibration and produce data at the end of calibration. In the
event of a failed calibration, the data will be printed with error codes and the system will alert the operator of the
failure. For information on error codes, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

TRACEABILITY

For Traceability information refer to the Calibrator instructions for use.

QUALITY CONTROL
At least two levels of control material should be analyzed daily. In addition, these controls should be run with each new
calibration, with each new reagent cartridge, and after specific maintenance or troubleshooting procedures as detailed
in the appropriate system manual. More frequent use of controls or the use of additional controls is left to the discretion
of the user based on good laboratory practices or laboratory accreditation requirements and applicable laws.
The following controls should be prepared and used in accordance with the package inserts. Discrepant quality control
results should be evaluated by your facility.

Table 1.0 Quality Control Material

CONTROL NAME SAMPLE TYPE STORAGE

TESTING PROCEDURE(S)
1. If necessary, load the reagent onto the system. A lot-specific parameter card must be loaded one time for each lot.
2. After reagent load is completed, calibration may be required.
3. Program samples and controls for analysis.
4. After loading samples and controls onto the system, follow the protocols for system operations.
For detailed testing procedures, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

CALCULATIONS
The SYNCHRON System(s) performs all calculations internally to produce the final reported result. The system will
calculate the final result for sample dilutions made by the operator when the dilution factor is entered into the system
during sample programming.

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REPORTING RESULTS
Equivalency between the SYNCHRON LX and UniCel DxC 600/800 Systems has been established. Chemistry results
between these systems are in agreement and data from representative systems may be shown.

REFERENCE INTERVALS

Each laboratory should establish its own reference intervals based upon its patient population. The following reference
intervals were taken from literature and a study performed on SYNCHRON Systems.7

Table 2.0 Reference intervals

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS
Literature Serum or Plasma (Male) 46 – 174 mg/dL 0.46 – 1.74 g/L
Serum or Plasma (Female) 46 – 142 mg/dL 0.46 – 1.42 g/L
SYNCHRON Serum or Plasma (Male) 51 – 153 mg/dL 0.51 – 1.53 g/L
Serum or Plasma (Female) 51 – 165 mg/dL 0.51 – 1.65 g/L

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS

Laboratory

Refer to References (8,9,10) for guidelines on establishing laboratory-specific reference intervals.

Additional reporting information as designated by this laboratory:

PROCEDURAL NOTES
ANTICOAGULANT TEST RESULTS

1. If plasma is the sample of choice, the following anticoagulants were found to be compatible with this method based
on a study of 20 healthy volunteers:

Table 3.0 Compatible Anticoagulants

LEVEL TESTED FOR IN VITRO AVERAGE PLASMA-SERUM
ANTICOAGULANT INTERFERENCE BIAS (mg/dL)
Ammonium Heparin 14 Units/mL NSIa
Lithium Heparin 14 Units/mL NSI
Sodium Heparin 14 Units/mL NSI
a NSI = No Significant Interference (within ± 10.0 mg/dL or 10%).

2. The following anticoagulants were found to be incompatible with this method:

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Table 4.0 Incompatible Anticoagulants
LEVEL TESTED FOR IN VITRO
ANTICOAGULANT INTERFERENCE PLASMA-SERUM BIAS (mg/dL)a
Potassium Oxalate/Sodium 2.0 / 2.5 mg/mL -30.3
Fluoride
a Bias is based on worst case instead of average. Plus (+) or minus (-) signs in this column signify positive or negative bias.

LIMITATIONS

None identified

INTERFERENCES

1. The following substances were tested for interference with this methodology:

Table 5.0 Interferences

SUBSTANCE SOURCE LEVEL TESTED OBSERVED EFFECTa
Bilirubin (unconjugated) Bovine 30 mg/dL NSIb
Hemoglobin RBC hemolysate 500 mg/dL NSI
Lipemia Intralipidc 400 mg/dL -8 mg/dL
a Plus (+) or minus (-) signs in this column signify positive or negative interference.
b NSI = No Significant Interference (within ± 10.0 mg/dL or 10%).
c Intralipid is a registered trademark of KabiVitrum, Inc., Clayton, NC 27250.

2. Refer to References (11,12,13) for other interferences caused by drugs, disease and preanalytical variables.

PERFORMANCE CHARACTERISTICS
ANALYTIC RANGE

The SYNCHRON System(s) method for the determination of this analyte provides the following analytical ranges:

Table 6.0 Analytical Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS
Serum or Plasma 35 – 300 mg/dL 0.35 – 3.00 g/L
Samples with concentrations exceeding the high end of the analytical range should be diluted with saline and reanalyzed.

REPORTABLE RANGE (AS DETERMINED ON SITE):

Table 7.0 Reportable Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS

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SENSITIVITY

Sensitivity is defined as the lowest measurable concentration which can be distinguished from zero with 95% confidence.
Sensitivity for ApoB determination is 35 mg/dL (0.35 g/L).

EQUIVALENCY

Equivalency was assessed by Deming regression analysis of patient samples to accepted clinical methods.

Serum or Plasma (in the range of 40.6 to 220.0 mg/dL):

Y (SYNCHRON LX Systems) = 0.943X + 7.6
N = 73
MEAN (SYNCHRON LX Systems) = 99.3
MEAN (Array) = 97.2
CORRELATION COEFFICIENT (r) = 0.9970

Serum or Plasma (in the range of 59 to 168 mg/dL):

Y (SYNCHRON LX Systems) = 1.006X + 1.7
N = 54
MEAN (SYNCHRON LX Systems) = 105.5
MEAN (NWLRL)a = 103.2
CORRELATION COEFFICIENT (r) = 0.9912
a Northwest Lipid Research Laboratories, Seattle, Washington
Refer to References (14) for guidelines on performing equivalency testing.

PRECISION

A properly operating SYNCHRON System(s) should exhibit precision values less than or equal to the following:

Table 8.0 Precision Values

1 SD CHANGEOVER VALUEa
TYPE OF
PRECISION SAMPLE TYPE mg/dL g/L mg/dL g/L % CV
Within-run Serum or Plasma 5.0 0.05 100.0 1.0 5.0
Total Serum or Plasma 7.5 0.08 100.0 1.0 7.5
a When the mean of the test precision data is less than or equal to the changeover value, compare the test SD to the SD guideline given above to
determine the acceptability of the precision testing. When the mean of the test precision data is greater than the changeover value, compare the
test % CV to the guideline given above to determine acceptability. Changeover value = (SD guideline/CV guideline) x 100.
Comparative performance data for a SYNCHRON LX System evaluated using the NCCLS Proposed Guideline EP5-T2
appears in the table below.15 Each laboratory should characterize their own instrument performance for comparison
purposes.

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Table 9.0 NCCLS EP5-T2 Precision Estimate Method
EP5-T2 Calculated
Test Mean
Point Estimates
TYPE OF No. No. Data Value
IMPRECISION SAMPLE TYPE Systems Pointsa (mg/dL) SD %CV
Within-run Serum Control 1 1 80 59.6 0.98 1.65
Serum Control 2 1 80 96.2 1.94 2.02
Serum Control 3 1 80 132.8 1.77 1.33
Total Serum Control 1 1 80 59.6 1.58 2.64
Serum Control 2 1 80 96.2 2.44 2.54
Serum Control 3 1 80 132.8 2.73 2.05
a The point estimate is based on the pooled data from one system, run for twenty days, two runs per day, two observations per run on an instrument
operated and maintained according to the manufacturer's instructions.

NOTICE
These degrees of precision and equivalency were obtained in typical testing procedures
on a SYNCHRON LX System and are not intended to represent the performance
specifications for this reagent.

ADDITIONAL INFORMATION
For more detailed information on UniCel DxC Systems, refer to the appropriate system manual.
Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are
trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries.
May be covered by one or more pat. -see www.beckmancoulter.com/patents.

SHIPPING DAMAGE

If damaged product is received, notify your Beckman Coulter Clinical Support Center.

REVISION HISTORY

Revision AF
Revised Quality Control section

Revision AG
Updated corporate address; removed EDTA as an Acceptable Anticoagulant claim.

Revision AH
Added Reagent Preparation visual aid to the Reagent Preparation section.

Revision AJ
Added Revision History

Revision AK
Added new language requirement: Czech, and Korean.

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Revision AL
Removed references to CX and LX systems as they are discontinued effective 12/2013.
Added Beckman Coulter trademark statement and disclaimer.

Revision AM
Added GHS Classification information

Revision AN
Added new language requirement: Romanian

Revision AP
Updates to comply with requirements per Beckman Coulter Global Labeling Policy.

Revision AR
Additional changes to comply with requirements per Beckman Coulter Global Labeling Policy.

Revision AT
Added new language requirement: Bulgarian, Serbian, and Vietnamese. Additional changes to comply with
requirements per Beckman Coulter Global Labeling Policy.

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SYMBOLS KEY

Table 10.0

Catalogue Number In Vitro Diagnostic

Contents Temperature limit

Manufacturer Expiration Date

Batch code Safety Data Sheet

CE Mark Consult Instructions for Use

Authorized Representative in the Date of Manufacture

European Community
Caution Biological risks

Reagent Cartridges Bar Code Card

Do not reuse

Made in USA of US and Foreign Components

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REFERENCES
1. Albers, J. J., Marcovina, S. M., "Standardization of Apolipoprotein B and A-1 Measurements", Clin. Chem., Vol.
35, No. 7 (1989).

2. Sternberg, J. C., "A Rate Nephelometer for Measuring Specific Proteins by Immunoprecipitin Reactions", Clin.
Chem., 23:1456 (1977).

3. Marrack, J. R., Richards, C. B., "Light-Scattering of the Formation of Aggregates in Mixtures of Antigen and
Antibody", Immunology, 20:1019 1040 (1971).

4. Tietz, N. W., "Specimen Collection and Processing; Sources of Biological Variation", Textbook of Clinical
Chemistry, 5th Edition, W. B. Saunders, Philadelphia, PA (2005).

5. National Committee for Clinical Laboratory Standards, Procedures for the Handling and Processing of Blood
Specimens Approved Guideline, NCCLS publication H18-A, Villanova, PA (1990).

6. CDC-NIH, Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, (Washington, D.C.: U.S.
Government Printing Office, 2009). (CDC 21-1112)

7. Tietz, N. W., Clinical Guide to Laboratory Tests, 3rd Edition, W. B. Saunders Company, Philadelphia, PA (1995).

8. National Committee for Clinical Laboratory Standards, How to Define, Determine, and Utilize Reference Intervals
in the Clinical Laboratory Approved Guideline, NCCLS publication C28-A, Villanova, PA (1995).

9. Tietz, N. W., ed., Fundamentals of Clinical Chemistry, 6th Edition, W. B. Saunders, Philadelphia, PA (2007).

10. Henry, J. B., Clinical Diagnosis and Management by Laboratory Methods, 22nd Edition, W. B. Saunders Company,
Philadelphia, PA (2006).

11. Young, D. S., Effects of Drugs on Clinical Laboratory Tests, 5th Edition, AACC Press, Washington, D. C. (2000).

12. Friedman, R. B., Young, D. S.,Effects of Disease on Clinical Laboratory Tests, 4th Edition, AACC Press,
Washington, D.C. (2001).

13. Young, D. S., Effects of Preanalytical Variables on Clinical Laboratory Tests, 3rd Edition, AACC Press,
Washington, D. C. (2007).

14. National Committee for Clinical Laboratory Standards, Method Comparison and Bias Estimation Using Patient
Samples Approved Guideline, NCCLS publication EP9-A, Villanova, PA (1995).

15. National Committee for Clinical Laboratory Standards, Precision Performance of Clinical Chemistry Devices
Tentative Guideline, 2nd Edition, NCCLS publication EP5-T2, Villanova, PA (1992).

Beckman Coulter Eurocenter S.A., 22, rue Juste-Olivier. Case Postale 1044, CH - 1260 Nyon 1, Switzerland
Tel: +41 (0)22 365 36 11

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