14.2.

CALIBRATION OF VOLUMETRIC GLASSWARE

Volumetric apparatus should be calibrated by the analyst before use in the laboratory.
Calibration is usually performed by weighing the amount of water delivered by or
contained in the volumetric apparatus. This mass is then converted to the desired volume
using the Density of Water:

Volume = mass / density

There are following types of glassware which are being used in analytical laboratory and
needs calibration.
(i) Volumetric flask
(ii) Pipette one mark /bulb pipette
(iii) Pipette graduated
(iv) Burette.
(v) Graduated measuring cylinders

Tips for Correct Use of Volumetric Glassware

Pipette
The Pipette is used to transfer a volume of solution from one container to another. Most
Volumetric Pipettes are calibrated To-Deliver (TD); with a certain amount of the liquid
remaining in the tip and as a film along the inner barrel after delivery of the liquid. The
liquid in the tip should not be blown-out. Pipettes of the "blow-out" variety will usually
have a ground glass ring at the top. And, drainage rates from the pipette must be carefully
controlled so as to leave a uniform and reproducible film along the inner glass surface.
Measuring Pipettes will be gradated in appropriate units. Once the pipette is cleaned and
ready to use, make sure the outside of the tip is dry. Then rinse the pipette with the
solution to be transferred. Insert the tip into the liquid to be used and draw enough of the
liquid into the pipette to fill a small portion of the bulb. Hold the liquid in the bulb by
placing your fore finger over the end of the stem.

Withdraw the pipette from the liquid and gently rotate it at an angle so as to wet all
portions of the bulb. Drain out and discard the rinsing liquid. Repeat this once more. To
fill the pipette, insert it vertically in the liquid, with the tip near the bottom of the
container. Apply suction to draw the liquid above the graduation mark. Quickly place a
fore finger over the end of the stem. Withdraw the pipette from the liquid and use a dry
paper to wipe off the stem. Now place the tip of the pipette against the container from
which the liquid has been withdrawn and drain the excess liquid such that the meniscus is
at the graduation mark. Move the pipette to the receiving container and allow the liquid to

1
flow out (avoiding splashing) of the pipette freely. When most of the liquid has drained
from the pipette, touch the tip to the wall of the container until the flow stops and for an
additional count of 10.

Burette
The Burette is used to accurately deliver a variable amount of liquid. Fill the burette to
above the zero mark and open the stopcock to fill the tip. Work air bubbles out of the tip
by rapidly squirting the liquid through the tip or tapping the tip while solution is draining.
The initial burette reading is taken a few seconds, ten to twenty, after the drainage of
liquid has ceased. The meniscus can be highlighted by holding a white piece of paper
with a heavy black mark on it behind the burette.

Place the flask into which the liquid is to be drained on a white piece of paper. (This is
done during a titration to help visualize color changes which occur during the titration.)
The flask is swirled with the right-hand while the stopcock is manipulated with the left
hand. The burette should be opened and allowed to drain freely until near the point where
liquid will no longer be added to the flask. Smaller additions are made as the end-point of
the addition is neared. Allow a few seconds after closing the stopcock before making any
readings. At the end-point, read the burette in a manner similar to that above. As with
pipettes, drainage rates must be controlled so as to provide a reproducible liquid film
along the inner barrel of the burette.

2
Volumetric Flasks
The Volumetric Flask is used to prepare Standard Solutions or in diluting a sample. Most
of these flasks are calibrated ‘To-Contain’ (TC) a given volume of liquid. When using a
flask, the solution or solid to be diluted is added and solvent is added until the flask is
about two-thirds full. It is important to rinse down any solid or liquid which has adhered
to the neck. Swirl the solution until it is thoroughly mixed. Now add solvent until the
meniscus is at the calibration mark. If any droplets of solvent adhere to the neck, use a
piece of tissue to blot these out. Stopper the flask securely and invert the flask at least 10
times.

Calibration of One Mark/Bulb Pipette

Requirements
• Calibrated balance
• Calibrated thermometer
• Dried beakers which have been previously rinsed with acetone
• Calibrated stop watch
Method:
Determination of volume
a) Clean the pipette thoroughly.
b) Fill up the pipette with distilled water to the mark. Care shall be taken to ensure that
there are no air bubbles inside the pipette.
c) Weigh the dried empty beaker (W1).
d) Deliver the water from the pipette to its maximum in the beaker(W2). The pipette
should be held in vertical direction and its tip should touch the beaker.
e) Weigh the beaker with water (W2).
f) Note the temperature of the distilled water using a calibrated thermometer.
g) Repeat the whole procedure till difference in two subsequent readings (W2 & W1)
differ not more than 0.01g.
h) Volume of the pipette is determined by using the following formula
V=M/D
Where V=Volume of the pipette in cm3
M=Weight of the water contained in the beaker obtained by the difference of the
weights W2 & W1 in gms
D=Density of water at 27oC i.e. 0.9965/gm/cc

Determination of Delivery time

a) Fill the pipette with distilled water to the maximum mark

3
 b) Ensure that there are no air bubbles in the pipette
c) Remove the finger from the jet and deliver the water from the pipette in the beaker.
Start the stopwatch simultaneously at the start of the delivery of water from the
mark on the pipette. Stop the stopwatch as the pipette has emptied at the maximum
delivery.
d) Repeat the procedure 3 to 4 times to determine reproducibility
e) Note the time of delivery in seconds and compare with limits as per nominal
capacity and type as per IS-1117-1975.

Precautions
a) The filled pipette should not contain any air bubbles.
b) The temperature of water should be measured after every reading using a calibrated
thermometer
o
c) Whole procedure of Calibration should be undertaken at 27 C
Reporting
• Report the actual volume of water contained in the pipette.
• Report the delivery time in seconds.

Volumetric Flask
Requirements
• Calibrated balance
• Calibrated thermometer
• Dried beakers which have been previously rinsed with acetone.

Procedure
a) Dry the flask under calibration, which has been previously rinsed with acetone.
b) Clean the flask thoroughly.
c) Weigh the dried flask empty (W1)
d) Fill the flask up to the graduation mark with distilled water.
e) Weigh the water filled flask under the same conditions (W2).
f) Note the temperature of the water using a calibrated thermometer.
g) Repeat the procedure 3 to 4 times to get reproducibility.
h) Volume of the flask is determined by using the following formula
Volume, V=W/ D
3
i) Where V=Volume of the flask in cm
j) W=Weight of the water contained in the flask obtained by difference of the weights
in W2 and W1.
D=Density of water at 27oC i.e. 0.9965/gm/cc
Precaution
a) The filled flask shall not contain any air bubbles.

4
 b) The temperature of water should be measured after every reading using a calibrated
thermometer with trace ability of NABL accredited agency.
c) No droplets of water should adhere to the inner surface of the walls of the flask
above the duration mark.
o
d) Whole procedure of calibration should be undertaken at 27 C.

Reporting
• Report the actual volume of water contained in the flask at 27oC.

Graduated Burette
Requirements
• Calibrated balance
• Calibrated thermometer
• Dried beakers which have been previously rinsed by acetone.
• Calibrated stop watch

Determination of Volume
a) Clean the burette thoroughly.
b) Fill up the burette with distilled water to the mark. Care should be taken to ensure
that there are no air bubbles inside the burette.
c) Weigh the dried empty beaker (W1).
d) Deliver the water from the burette into the weighed beaker (W1) from 0 mark to its
maximum capacity.
e) If required the delivery of water may be regulated at 5 different points, for example
50ml burette, maximum points may be 10ml, 20ml, 30ml, 40ml and 50ml.
f) While delivering the water the burette should be held vertically and tip should touch
the beaker.
g) Weigh the beaker with water (W2).
h) Note the temperature of the water by a calibrated thermometer
i) Repeat the procedure till the difference in two subsequent reading (W2 and W1)
differ not more than 0.01gm.
j) Volume of the burette is determined by using the following formula.
Volume, V=M/D
Where V=Volume of the burette in cm3
M=Weight of the water contained in the beaker obtained by difference of W2
and W1 in grams.
D=Density of water at 27oC i.e. 0.9965/gm/cc
Determination of Delivery Time
a) Fill the burette with distilled water beyond the 0-ml mark.
b) Ensure that there are no air bubbles in the burette

5
 c) Loose the burette knob to its maximum limit and hold the lower tip of the burette
with finger.
d) Remove the finger, the moment lower meniscus reaches 0-ml mark and start the
stopwatch.
e) Stop the stopwatch when the lower meniscus reaches its maximum capacity mark.
f) Note the time of delivery.
g) Repeat the procedure 3 to 4 times to determine its reproducibility.
Precautions
a) The filled burette should not contain any air bubbles
b) Temperature of water should be measured after every reading using a calibrated
thermometer with traceability to an NABL accredited agency.
o
c) Whole procedure of calibration should be undertaken at 27 C.
Reporting
• Report the actual volume of water contained in the burette.
• Report the delivery time in minutes.
Graduated Pipette
Graduated pipette is very similar to graduated burette, both in design and its use. So
the calibration can be done using same method described above.
Graduated measuring cylinder
Requirements
• Calibrated balance
• Calibrated thermometer
• Dried beakers which have been previously rinsed with acetone.

Procedure
a) Dry the measuring cylinder under calibration, which has been previously rinsed with
acetone.
b) Clean the measuring cylinder thoroughly.
c) Weigh the measuring cylinder empty (W1).
d) Fill the measuring cylinder with distilled water up to the maximum graduation
mark*.
e) Weigh the water filled measuring cylinder under the same condition (W2).
f) Note the temperature of the water using a calibrated thermometer.
g) Repeat the procedure 3 to 4 times to get reproducibility
h) Volume of the measuring cylinder is determined by using the formula:
Volume, V=W/D
Where V = Volume of the measuring cylinder in cm3
W = Weight of the water contained in the measuring cylinder obtained by
difference of the weights W2 and W1.
D = Density of water at 27oC i.e. 0.9965/gm/cc

6
Precautions
a) The filled cylinder shall not contain any air bubbles.
b) The temperature of water should be measurement after every reading using a
calibrated thermometer with trace ability of NABL accredited agency.
c) No droplets of water should be adhering to the inner surface of the walls of the
cylinder above the graduation mark.

Reporting
• Report the actual volume of water contained in the measuring cylinder at 27oC.

*Note: More points should be selected to cover full range of the cylinder depending on its
use, at least 3-5 points. For example a 50ml may be calibrated at 10ml, 20ml, 30ml, 40ml
and 50ml points.

References:

¾ Published specification in accordance with which the tests are performed : BIS
Standards
¾ IS: 915-1997, one mark volumetric flasks
¾ IS: 1117-1975, one mark pipettes
¾ IS: 4162-(part I & II)-1985, Graduated pipettes & graduated burette
¾ IS: 875-1975, Graduated measuring cylinders
¾ IS: 9440-1979, Laboratory balances
¾ IS: 9865-1981, Laboratory standard weights
¾ IS: 14012-(Part I)-1993 Quality Assurance requirements for measuring equipments.
¾ IS 8897 –1978 Tables for calibration & method of verification of volumetric
glassware

Author: Dr. Robin Kumar,

Senior Scientific Officer,
Indian Pharmacopeia Commission,
Ghaziabad.
Email: [email protected]

-------------------------------

7
 15. GENERAL REQUIREMENTS FOR THE COMPETENCE OF
TESTING AND CALIBRATION LABORATORIES
Note: Compliance of GLP norms is a statutory requirement for approved drugs testing
laboratories and the in-house laboratories of manufacturing units. Compliance of GLP
norms could enable a laboratory for NABL accreditation or other international
accreditations specified hereunder also with such additional requirements as may be
needed.The requirements for such accreditations are given hereunder for the guidance of
the laboratories.
Foreword:

ISO: International Organization for Standardization

IEC: International Electrochemical Commission
CASCO: ISO Committee on conformity Assessment Responsible for the development of
international standards and guides.
ISO/IEC 17025 was prepared by CASCO and through Circulated for voting to the
national bodies of both ISO and IEC and approved by both organizations.

Introduction:
The laboratories (Testing/Calibration) have to meet all the requirements of this standard
in case they wish to demonstrate that:
• They are operating under a sound management system
• They are technically competent
• They are able to generate technically valid data and results
• Accreditation bodies such as NABL are using this international standard as a basis
for their accreditation to recognize the competence of testing and calibration
laboratories
• The testing and calibration results are accepted between the countries which have
entered into a mutual recognition agreement with equivalent bodies through
APLAC/ILAC
Scope:
• Specifies the general requirements only
• Covers testing and calibration performed by using standard/non-standard and
laboratory developed methods
• Applicable to all organizations performing tests / calibrations/sampling

8
 • Includes laboratories where testing or calibration forms part of inspection or
product certification
• Applicable to all laboratories regardless to the number of personnel's or extent of
scope.
• Requirement of clauses are applicable as per the activities covered.
• Applicable to first, second or third party testing
• May be used by customers, regulatory and accreditation bodies to recognize the
lab. competence
• Not intended to be used as a basis for laboratory certification
• Regulatory and safety requirements are not being covered by this standard.
• Compliance to the requirement of the standard correlates with the compliance to
the principles of ISO 9001.
• Being used by the laboratories to develop their management system for quality,
administrative and technical operations.
Compliance to relevant clauses:
• Sound management system
Clause-4, Management requirements
• Technical competency
Clause-5, Technical requirements addressed in Quality Manual for compliance
• Compliance output:
Generation of technically valid data and results
Management Requirements:
• Organization
• Management system
• Document control
• Review of request tender and contracts
• Subcontracting of test and calibrations
• Purchasing services and supplies
• Service to the customer
• Complaints
• Control of nonconforming testing and /or calibration
• Improvement
• Corrective action
• Preventive action
• Control of records
• Internal audits
• Management reviews
Technical Requirements:
• General
• Personnel

9
 • Accommodation and environmental conditions
• Test and calibration methods and method validation
• Equipment
• Measurement traceability
• Sampling
• Handling of test and calibration items
• Assuring the quality of test and calibration results
• Reporting the results
Management Requirements:
Laboratory/Organization:
• Legal responsibility
• Able to satisfy the needs of customer, regulatory authorities and recognizing
bodies.
• Has facility (permanent, site, temporary and mobile )and resources to carry out
work
• Stating conflict of interest, if any
• Managerial and technical personnel
• Defined authority and responsibility of key personnel
• Well defined organization & management structure
• Adequate supervision of testing and calibration staff
• Quality manager and technical manager/technical management system
• Deputies or key managerial personnel
• Free from undue internal, external, commercial, financial or other pressures
• Protection of customer confidential information and proprietary rights
• Ensure competence, impartiality, judgment and operational integrity
• Appropriate communication process
Management System:
• Shall establish/develop, implement and maintain a management system
appropriate to scope
• Documented policies, objectives, systems, programs, procedures and instructions
• Documentation shall be communicated to, understood by, available to, and
implemented by appropriate personnel
• Documented quality policy statement in quality manual (Clause 4.2.2 a to e)
• Establishment of overall objectives
• Reference to supporting procedures in quality manual (system/technical)
• Level of documentation structure (three/four)
• Role and responsibility of quality manager and technical management system
• Commitment to meet customer, regulatory and statutory requirements
• Improving the effectiveness in continual manner
Management System commitment:

10
 Establishment, implementation and maintenance of system with well documented
policies, objectives, programs, procedures and instructions being followed all the
times as per well defined role and responsibilities of staff. These are subjected to
periodic reviews and audits in terms of ensuring a firm commitment for its
continual improvement through appropriate personnel at all levels of
organizational hierarchy.
Document Control:
Procedure to control both internal and external documents being maintained in the
laboratory
1. Document approval and issue :By authorized personnel in terms of its unique
identification, availability, revision and assurance for unintended use of obsolete
documents
2. Document changes: in terms of review and approval with identification of altered
text both in hard copies or in computerized system
Requests, Tenders & Contracts Review:
The laboratory shall ensure
1. Methods and other requirements are adequately defined and documented
2. Capability and resources to meet the requirements
3. Capability of selected methods in meeting the customer requirements

The review should:

• Be acceptable to both laboratory and customer
• Be maintained in terms of records
• Cover the details in terms of subcontracted work
• Able to provide customer of any deviation from the contract
• Being liable to be amended at any stage of work
• Have provision for communication to all affected personnels in terms of its
amendments

Subcontracting of Tests and Calibrations:

Due to unforeseen reasons or on continuous basis
1. With competent sub contractor that complies with the standard such as ISO/IEC
17025:2005
2. Information to and approval from customer
3. Responsibility towards customer if subcontractor is not specified from customer
4. Maintenance of records.

Purchasing Services and Supplies:

Establishment of policy and procedures for:

11
 1. Purchasing of services/supplies including reagents and consumables
2. Inspection and verification of critical consumables against specific requirements
3. Evaluation of the suppliers of critical consumables
4. Maintenance of relevant records including list of approved vendors

Service to the Customer:

The laboratory shall:
1. Be co-operative with customers
2. Ensure confidentiality
3. Provide limited access to lab. for witnessing tests
4. Seek feedback from customers
5. Use customer information as a management tool for improvement
Complaints:
Establishment of policy and procedures for
1. Receiving of complaints
2. Resolution of complaints in terms of investigation and corrective actions taken
3. Maintenance of complaint records.

Control of Non Conforming Testing and/or Calibration Work:

Implementation of established policy and procedures at a time when
1. Any aspect of testing/calibration or
2. Results of testing
(Do not conforms to its own procedures or to the agreed requirements of its customer)

The system shall ensure that:

1. Responsibility and authority for the management of non-conforming work is
designated both in terms of halting and resumption of work
2. Pre-defined actions such as evaluation, correction, corrective actions are taken as
and when the activity is defined
3. Recalling of work and informing to customer, if required.

Improvement:
• Shall continually improve the effectiveness of management system through the
use of
• Quality policy
• Quality objectives
• Audit results
• Analysis of data
• Corrective and preventive actions
• Management review

12
Corrective Actions:
Implementation of established policy and procedures in terms of designated authority for
taking corrective actions at a time when any non conformance is identified in
management system or technical operations.

Procedural Steps:
1. Root cause analysis
2. Selection and implementation of corrective action
3. Monitoring of corrective action
4. Conducting additional audits, if required

Preventive Action:
• Comprises of following steps:
1. Identification of potential source of non-conformity
2. Evaluation for the requirement for preventive action
3. Development, implementation and monitoring of action plan

Control of Records:
• Quality and technical records
• The procedure comprises of:
Identification, collection, indexing, access, filing, storage, maintenance and disposal of
records
• Compliance criteria for records:
1. Shall be legible
2. To be readily retrievable
3. Stored in an environment to prevent damage/deterioration or loss
4. Retention time should be established
5. To be held secure and in confidence
6. Protection to electronically stored records in terms of back up and unauthorized
access

Technical records:
• Original observations, derived data, calculations, calibration certificates, staff
records, test report copies, work sheets, work books, control graphs and feedbacks
etc.
• Recording of observation data and calculations
• Rectification of mistakes occurring in the record
• Measures taken for electronically stored records

13
Internal Audits:
• Conducted as per plan ,schedule and procedure
• Being conducted periodically
• To verify management system requirements by addressing all elements including
test activities
• Responsibility lies on quality manager
• Through trained and qualified personnel
• Independent of the activities to be audited
• All activities are to be covered in one year cycle
• Recording of audit findings in terms of observations related to area of activity
audited and non-conformances recorded
• Proposing and taking corrective actions
• Verification and recording of corrective actions in terms of time frame
implementation in effective manner
Management Reviews:
• To be held periodically as per schedule and procedure
• Conducted from top management
• Minimally conducted once in 12 months
• Review of system and testing/calibration activities
• Recording of review findings and actions to be carried out within agreed upon
time scales
The agenda includes the review of:
• Policy and procedures
• Report from management/supervisory personnel
• Audit outcomes/external bodies assessment
• Corrective and preventive actions
• Inter lab comparison/P.T programs results
• Changes in volume and type of work
• Customer feedback
• Customer complaints
• Recommendation for improvement
• Quality control activities
• Management of resources
• Staff training

The responsibility lies on Quality manager

Technical Requirements:

General

14
Factors that determine the correctness and reliability of tests and/or calibrations.

Human factor, Accommodation and environmental conditions, Test and calibration

methods, Method validation, Equipment, Measurement traceability, Sampling and
handling of test and calibration items. Etc.
• These factors are to be controlled to the extent of their minimum contribution
towards total uncertainty of measurements
• Taken into account in the development of test and calibration methods and
procedures
• staff training and authorization
• Selection and calibration of equipment etc.

Personnel
The management shall ensure:
1. Competence of personnel at all levels in terms of performing routine and specific
tasks
2. Appropriate supervision for the staff under going training
3. Having policy and procedure to identify training needs and providing training
4. The relevance of training program in terms of present and anticipated tasks
5. Evaluation of training actions taken

The management shall:

1. Use employed or contractual persons
2. Ensure the competence of contractual person
3. Ensure the adequate supervision of contractual person
4. Ensure their working in accordance to the laboratory management system

The management shall:

1. Maintain current job description of laboratory staff
2. Authorize specific personnel to perform specific/particular tasks
3. Maintain records of relevant authorization
4. Maintain personnel records pertaining to qualifications, training, skills and
experience of staff working in the laboratory.

Accommodation and Environmental Conditions:

The laboratory shall:
1. Create conditions to facilitate the correct performance of tests and/or calibrations
2. Ensure that environmental conditions do not adversely effect the required quality
of any measurements
3. Ensure to take particular care in case activities are being undertaken at sites

15
 4. Ensure to document those technical requirements that effect the results of tests
and /or calibrations

The laboratory shall:

1. Monitor, control and record environmental conditions appropriate to the technical
activities concerned
2. Ensure the effective separation between incompatible activities
3. Ensure the controlling of access to the area affecting the quality of tests
4. Ensure good housekeeping by taking relevant measures (both routine and specific)

Test, Calibration Method and Method Validation:

The laboratory shall ensure:
1. The use of appropriate methods and procedures for all tests and/or calibrations
2. To have instructions on the use and operation of all relevant equipments and
handling/preparation of test items .
3. Use of latest valid edition of international/national standard test and or calibration
methods including method for sampling
4. The laboratory shall ensure the confirmation of proper operation of standard
methods

5. The validation of laboratory developed/ designed methods, non-standard methods

and standard methods used outside of their intended scope, in appropriate manner
before use

6. The validation is as per the necessity to meet the need of given application

Uncertainty of Measurement:
The laboratory shall:
Have and apply a procedure to estimate the uncertainty of measurement for all internal
calibrations and tests by taking into account all the significant/important component
affecting the test/calibration activity. Estimation depends upon nature of
method/customer requirement and existence of narrow limits on which decision on
conformity to a specification are based.

Control of Data:
The laboratory ensures:
1. Appropriate checks in systematic manner for calculations and data transfer
2. The maintenance of computers and automated equipments being used for
acquisition, processing, recording, reporting, storage or retrieval of test or
calibration data. The provision of environmental and operating conditions
necessary to maintain the integrity of test and calibration data

16
Equipments:
The laboratory shall ensure:
1. The furnishing with all items of sampling, measurement and test equipment
required for correct performance of tests and /or calibrations
2. The capability to achieve the required accuracy and shall comply with
specifications relevant to the tests and /or calibrations concerned

The laboratory ensures:

1. The calibration of all the relevant equipments
2. The operation by authorized personnel
3. The availability of up to date instructions
4. Unique identification of all the equipments
5. Maintenance of equipment records/history

The laboratory ensures:

1. To have procedures for safe handling, transport, storage, use, and planned
maintenance of measuring equipments
2. To have calibration status of all the equipments with proper updating of correction
factors
3. The performance of intermediate checks to maintain confidence in calibration
status
4. Safe guard from adjustments which would invalidate the test and /or calibration
results

Measurement Traceability:
The laboratory shall ensure:
1. That all equipments used for tests and /or calibrations and sampling and having
significant effect on accuracy and validity of results are to be calibrated before
being put into service.
2. Having a program and procedure for ongoing calibration of its equipments

• For calibration laboratories the program for calibration of equipment shall

be designed and operated to ensure that calibrations and measurements
made are traceable to the international system of units (SI).
• For testing laboratories the above mentioned requirement is applicable for
measuring and test equipments
• Where traceability of measurement to SI unit is not possible/relevant
certified reference material, agreed methods or consensus standards are
required

17
Reference Standards/Materials:
The laboratory shall ensure:
1. Having program and procedure for the calibration of its reference standards and
shall be calibrated from a body that can provide traceability
2. Generally using the reference standard for calibration only
3. Where possible, the traceability of reference material to SI units or to certified
reference material. Checking of internal reference material as practicable.
4. Maintaining calibration status of reference, primary or working standards material

The laboratory shall ensure:

1. The checks needed to maintain confidence in the calibration status of reference,
primary, working standard and reference material shall be carried out according to
defined procedure and schedules
2. Having procedure for safe handling, transport, storage and use of reference
standards and reference material to protect their integrity and prevent their
contamination or deterioration.

Sampling:
• Establishment of plan and procedures
• Its availability at locations where sampling is undertaken
• Being based upon appropriate statistical method
• Recording of deviations, addition or exclusions from sampling procedure as per
customer requirements
• Procedure to record data and operations related to sampling if it forms part of
testing

Handling Test & Calibration Items:

• Shall have procedures for transportation, receipt, handling, protection, storage,
retention and disposal of test and calibration items
• Shall have system for its identification
• Proper condition of test item on its receiving
• Shall have procedure and facilities to avoid deterioration, loss or damage to test
and calibration items
• Retention and disposal as per norms.

Assuring Quality of Test and Calibration Results:

The laboratory shall ensure:
1. Having procedures for monitoring the validity of tests and calibrations being
taken up by the lab

18
 2. The planning and review of monitoring in terms of generation of quality control
data
3. Taking action in case analyzed data is found to be outside the pre-defined criteria

The laboratory shall ensure the monitoring through:

1. Regular use of certified/secondary reference material
2. Participation in ILC/PT programme
3. Replicate testing using same or different methods/analyst and retesting of retained
items
4. Correlation of results for different characteristics of an item
5. By any other appropriate means

The laboratory shall ensure:

1. Expressing opinion and interpretation in test report by documenting the basis for
the same
2. The identification of sub contractor on the report.
3. Electronic transfer of results in a manner to meeting of this standard requirements
4. Designing of standard report formats
5. Issuing of amended test report for any material amendment in initial report by
meeting the requirement of this standard.

Author: Mr. S.K. Gaind,

Techno-Management Consultant
Email: [email protected]
---------------

Note: For other requirements such as procedures for making applications, fee payable etc, the usrers are
advised to contact the certification agency concerned or visit their web-sites.

16. PRACTICAL RISK-BASED SYSTEMS APPROACH

Quality Assurance in QC Labs

Topics:
 Additional to GCLP and GMP
 ICH Q 10 parameters
 Process performance
 Product lifetime quality
 ‘Process’ is all about the QC-related activities
 ‘Product’ is final test report with complete raw data

19
Getting Started:
 To begin, list all of the items required to be sampled and tested
 Alter the list whenever necessary
(A) Starting and packaging materials
(B) In-process testing
(C) Finished goods
 To get on with a systems approach:
Systems:
1. Sample management system
2. Testing process monitoring system
3. System of controlling ‘Product’ quality and state of control
4. Quality risk management system
5. Change management system for continual improvement and innovation through
audits and CAPA
6. Out of specification Investigation records
Systems: continued:
 Calibration Lifecycle Management system
 Reference materials management system
 Analytical Methods Validation Management system
 Go through each system and risk management for what can go wrong
Sample Management System:
 Prepare SOPs for all of the activities connected with sampling and sample
handling
 Sampling
 Qualification of samplers by a training program duly recorded and keep random
checking with records
 List of trained samplers
 Sampling plan:
(A) Individual containers
(B) Pooling
Risk Management:
Involve workers and chemists actually carrying out the activities, in preparation of
the SOP
 Mix-up of similar-sounding items
 Cross contamination
 Assign code numbers
 Store in separate areas
 Good design and maintenance of the sampling booth and equipment
Reverse-flow laminar

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  Clean sampling tools. Appropriate for some specific items. Eg for aseptic
sampling, crude drugs etc.
 Identify and list items where aseptic sampling occurs; list any precautions
required, and display near SOP
 Closure of the sampled containers
 Identify more risks according to your own area, facilities, and operators
Watch them operate
Think about what can go wrong and how to prevent it
 Handling, proper labelling, and secure transport to the lab
 Provide locked main containers
 Entry in registers/computer systems
 Store in closable drawers or cabinets labelled on the outside

Testing Process:
 Prepare a day’s work schedule in the lab, preferably on the previous evening
 Continue the incomplete testing of samples after checking the condition in which
they were previously left
 Allocate new samples to qualified analysts with analytical work sheets and
complete instructions
 As the tests are being done, record all of the readings and observations online at
the same time they are checked and signed by the analyst and the checker

Testing Process Monitoring:

 Review records, calculations, chromatograms, and graphs; signature and with date
and time.
 Prepare the report
 Send report or information to whoever has sent the sample with opinion and
signature
 Keep control samples

Risk Management:
 Avoid delays by preplanning and constant checking by group leaders/senior
analysts
 Keep the valid equipment, reagents, and reference materials ready in advance
 Correct non-conformances as soon as possible
 Attach weight records, chromatograms, graphs, etc., to the raw data
 Review on the same day and sign by the designated persons
 Return standards to the appointed place
 Do not return any standard material to the container
 Prepared lists of dos and don’ts exhibited, implemented

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  Do what is written and write what you do
 ID code no. of equipment and of reference materials inserted in raw data with
calibration due and use-before dates
 Chromatograms and graphs are sequentially numbered
 Corrections, if any, suitably crossed out, still legible, signed, and dated with time
 Balances suitably cleaned after each operation
 Column washing
 Non-conformances noted and corrected
 Authorized change control
 Integrity preserved for incomplete tested samples
Risk Assessment:
 Records of use of reference materials and reagents completed
 Safety precautions observed
System of Product Quality:
Control and State of Control
 Product is test report with complete raw data
 Unique identification number of the report
 Description including packaging
 Receipt of the sample and sampling date
 Sample identification
Product Quality Control:
 Deviation, if any, from the approved methods in the report
 Results of each test in details
 Opinion and interpretation
 Guidance for improvements, if any
 Uncertainty measurement, if required
 Date of completion and signature with name
Risk Management:
 Check transfer of results from raw data
 All the necessary signatures in the raw data including on the chromatograms,
graphs, etc., which are attached
 Clearly visible contents of the report
 Good quality paper for long-term archiving
Risk Management System:
 Establish an Analytical Quality Assurance (AQA) team
 Assign responsibilities
 Regularly review and update the specifications set by Quality Control
 Approve Change Control
 Institute CAPA
 Audit and improvements

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Risk Management:
 Must have a trained deputy for key personnel
 Not to carry out any tests by the checkers
 Continually monitor the process of testing with record with time and date
 Approve preferably on the same date the approval is sought
 Interact with the analysts with or without QC managers
 THERE MAY BE SPECIFIC ACTIVITIES IN CERTAIN LABS. PREPARE A
FLOW CHART FOR ALL THE ACTIVITIES

Author: Dr. J L Shipahimalani,

Email: [email protected]

------------------------

17. CHALLENGES IN COMPLIANCE OF PHARMACOPOEIAL

STANDARDS

Quality is Invisible when GOOD but impossible to ignore when BAD

WHAT IS QUALITY? It is simply a assemblage of knowledge, information & data.

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To achieve it one has to follow the 5 M’s of Quality:
` Man
` Motivation
` Material
` Machinery
` Manuals/Methodology ( SOP)

FACTORS INFLUENCING QUALITY:

Right sampling Right Analysts Documentation

Right labelling Right Environment Interpretation

Right Quantity Right reagents Turn around time

Right Storage Right Instrument Report to right user

Right Selection of method

HOW PREPARED IS YOUR LAB FOR TESTING OF DRUGS?

• Are you comfortable working in your laboratory in the hot summer days?
• Is your HPLC instrument working comfortably when your AC machine is down?
• Do you have a safe working environment your chemical laboratory?
• Are your samples protected from heat and humidity when under analysis or before
being taken up for testing?
Besides the normal qualification of equipment, glass wares and chemicals, environment
plays a vital role in safe testing .No specific guideline is given for controlling the
environmental condition nor is any specified limit prescribed for validating it.
WHO mentions only that the” laboratory should ensure that the environmental
conditions are monitored, controlled and do not invalidate the results or adversely
affect the quality of measurement results.”
The Pharmacopoeia specifies in its “ General Chapter “ that Assay & Tests are to be
carried out at a temperature between 20°c to 30 °c and measurements at 25° c while
instruments require to be installed at ambient temperature of 25° c and a humidity range
of 45 % to 60 % is permitted for testing. After the recent implementation of Schedule L1,

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the requirement for premises is possibly the most discussed topic as it involves a good
investment in construction and installation.
It specifies that:
• Air ventilation system should ensure dust free environment with adequate
ventilation.
• If necessary air-conditioning to maintain satisfactory temperature and
relative humidity that will not adversely affect the testing and storage of
drugs or the accuracy of the functioning of the laboratory equipments or
instruments

• The use of chemicals and other potentially hazardous compounds separates

laboratories from other types of building spaces.

• The requirement for laboratory room air conditioning is not alike and differs from
laboratory to laboratory depending on different hazard levels and uses.

• Some specified laboratories may be controlled by a complete package of heating,

ventilation and air conditioning (HVAC) while others can be adjusted with room
AC s only.

• Room AC can be installed in the Instrument Laboratory where sophisticated

analytical instruments are installed with no much requirement to supply fresh
cooled air and there is no exhaustion of large quantities of air from inside.

• A wet chemistry lab having a number of fume hoods, large volume of air is
regularly extracted from the room and additional heat is generated from the ovens,
incubators and other heating units. In that case regular split or windows A/C
cannot give the required cooling effect .The mixing of air should not be permitted
and proper ventilation should be maintained to allow an adequate mixture of
gases in the air we breathe , control odour and remove contaminants from
occupied space.

• The Microbiology Laboratory would require an additional Clean room facility

where the concentration of airborne particles should be controlled to meet a
specified cleanliness class by constantly re-circulated by HEPA filters through
HVAC system from outside should be filtered to exclude dust and the air entering
a clean room is filtered to exclude dust

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RELIABILITY OF ANALYTICAL
DATA:

The data generated from the analytical

instruments should reliable and
accurate. This depends on the
following factors :

` Validation of analytical method

` Reliability of the instruments
used
` Proper training of the analysts

INSTRUMENT QUALIFICATION:
` Equipment qualification is the way to ensure that the analytical equipment is fit to
do what is expected to do. It is important if analytical methods are transferred
from laboratory to laboratory, for example in the transfer of a gradient elution
method in HPLC the gradient delay volumes may influence retention time and
selectivity of the separation.
It depends on the principle of 4 Qs
` Design Qualification
` Installation Qualification
` Operational Qualification
` Performance Qualification + System Suitability Test

DESIGN QUALIFICATION:
` 1. HPLC system with a variable UV Detector, isocratic pump and auto sampler is
enough for routine assay of main active ingredients.
` However if the HPLC system is intended to be used for impurities assay system
then a system with a gradient pump is required.
` So Design Qualification (DQ) should be do it yourself(DIY ) prepared in
consultation with Original Equipment Manufacturer (QEM)

OPERATIONAL QUALIFICATION (OQ):

` Operational Qualification should be checked according to the user’s requirement.
` A spectrophotometer itself or the UV-Vis detector of HPLC unit is usually
qualified with Holmium oxide solution in 10 % perchloric acid covering a range
of 241 -273 nm .

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 ` But what about the range 210 nm to 220nm which is frequently used in assay.
This requires to be qualified with caffeine which has two absorption bands at 205
and 273 nm and so both standards can be used in combination to quantify the
detector over the range of 205-641 nm (or to 361 nm for the UV range only )

PERFORMANCE QUALIFICATION
` The end user is responsible for the PQ qualification.
` Once the Equipment qualification has been started, there is no stopping the
process.
` HPLC pump seals can wear and spectrophotometer lamps have a finite life. The
analyst must be aware of the critical instrument parameters to be monitored as
part of a routine process
` A pH meter is not really measuring the acidity but the potential between the
electrode and the solution and then calculated into pH .Its detector measures the
concentration of hydrogen ions as a potential and calculates this to pH – ph = log [
H+)
` So the pH meter is a Voltmeter using high resistance electrode and is calibrated
by dipping the electrodes into buffers and adjusting the electrodes.
` The qualification of the pH meter will include qualifying the temperature
accuracy, voltage accuracy, pH accuracy signal noise ratio and a holistic accuracy
test for the complete instrument

PERIODIC CALIBRATION:
` Performance qualification is verified by periodic calibration of each module.
` Specific performance is checked by using certified calibration standards of known
traceability.
` System suitability test is performed to verify the instrument, reagents, columns
and analysts to check a specific method.
` It establishes a equivalence of a established method
` suitability tests are performed for a specific method to determine its effectiveness
and specific criteria as :
` Column Efficiency ( Plate count )
` Resolution
` Tailing factor
` Capacity factor

CHOOSE THE RIGHT COLUMN

` Most HPLC test procedure are based on Reverse phase C18 columns mention as
L1 column in USP.

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 ` There are about 600 different brands of C18 column which differ from
manufacturer to manufacturer based on nature of the packing material based on
chain length, end caooing, base –deactivation, pore size and specific surface area.
It becomes insufficient to select a suitable column for analysis.
THE MIS-UNDERSTOOD LABORATORY SOLVENT – REAGENT WATER
FOR HPLC:

` Reagent water is the most widely used analytical laboratory reagent in HPLC
analysis.
` Though chromatographers take care about the purity of standards, organic
solvents and other mobile phase components, the quality of water is taken for
granted.
` Reagent water affects nearly every aspect of HPLC analysis from sample and
standard preparation to column rinsing and elution.
` Water is the largest reagent consumed in terms of volumes

` Poor quality water reduces chromatographic performances by affecting resolution

and integration, introducing ghost peaks, altering stationary phase selectivity and
irregular base line.
` The TOC level affects the chromatographic peaks. TOC level above 50 ppm
affects chromatographic analysis.
` At 210 nm UV wavelength any organic contaminant containing a carbonyl
functional group will be absorbed, while they would not at 254 nm. As many LC
methods utilize 210 nm wavelengths; water containing such organic contaminant
should be avoided.
` HPLC bottled water or distilled water that has been standing in a laboratory
environment for more than 8 h should be avoided for HPLC analysis.
` Freshly prepared water should be used for HPLC analysis.

TEST METHOD TO BE USED

` Analytical method used for testing should be either compendia l as per
Pharmacopeia or published validated method as per ICH guidelines or in- house
validated method.

“IF you have not documented it you have not done it”
Laboratory records
` Description and identification of sample received
` Description of method of testing
` Record of all data secured in the course of the test

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 ` Record of test results and how they compare with standards of identity, strength
and quality
` Record of all deviations and modification of test
` Record of standardization of reference standards
` Record of calibration of equipments

RECORD KEEPING
` Laboratory records shall include complete data derived from all tests necessary to
assure compliance with established specifications and standards, including
examinations and assays, as follows:
` A description of the sample received for testing with identification of source (that
is, location from where sample was obtained), quantity, lot number, date sample
was taken, and date sample was received for testing.
` A statement of each method used in the testing of the sample. The suitability of all
testing methods used shall be verified under actual conditions of use.
` A complete record of all data secured in the course of each test, including all
graphs, charts, and spectra from laboratory instrumentation, properly identified to
show the lot tested.

Documentation of problems:
Develop a problem log record for each piece of equipment
x Date problem occurred, removed from service
x Reason for breakdown or failure
x Corrective action taken
x Date returned to use
x Change in maintenance or function check

Benefits of a Maintenance Program:

` Greater confidence in the results
` Safety
` Fewer interruptions of work
` Lower repair costs
` Elimination of premature replacement
` Reduction of variation in test result

QUALITY ASSURANCE IS RESPONSIBILITY OF ALL STAFFS

LET US TUNE TOGETHER FOR A BETTER TOMORROW
Author: Dr. Gopa Ghosh,
Retd. Director,
CDTL,Mumbai.
Email: [email protected]

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 -------------------------

18. STANDARDS OF HERBAL DRUGS IN IP

Standards for herbals-Scenario

„ Herbals/Naturals
‰ Approved as Drugs-Very few
‰ ASU Drugs/Medicines-Large numbers, Different regulations,
API’s/AFI’s.
‰ Food (Herbal) Supplements- ?? [ Area needs attention]
‰ Cosmetics with herbs?? [Area needs attention]-IS Standards “General
Guidelines for Herbal Cosmetics, IS:
„ Overall Area loosely regulated-Inadequate availability of Quality Specifications.

Role of IP - IN DEVELOPING QUALITY MONOGRAPHS FOR HERBS AND

HERBAL PRODUCTS FOR PHARAMCOPEIA

„ Journey so far-IP 2010

„ Examples- features, coverage, feed backs
„ Challenges
‰ New Science and Tools under Evaluation
ƒ DNA based testing
ƒ Image analysis techniques.
„ Infrastructure
„ Skills and Competency
„ Forecast of needs
„ Mind set of regulators.
„ Acknowledgements
„ Way Forward

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31
Date

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