2024

Guidance on
Stability Studies of
In-vitro Diagnostic
Medical Device
(IVDMD)

Central Drugs Standard Control Organization

Directorate General of Health Services
Ministry of Health and Family Welfare
Government of India
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India

Central Drugs Standard

Control Organization
(In-Vitro Diagnostic Division)

Guidance Document

Title : Guidance on Stability Studies of In-

Vitro Diagnostic Medical Device
(IVDMD)

DocNo. : CDSCO/IVD/GD/Stability/02/2022

Date : 05.04.2024

Notice:

This Guidance document is aimed only f or creating public awareness about In-Vitro Diagnostic
Devices Regulation by CDSCO and is not meant to be used for legal or professional purposes. The
readers are advised to ref er to the statutory provisions of Medical Devices Rules, 2017 and
subsequent amendments and clarifications issued by CDSCO time to time f or all their prof essional
needs.

CDSCO/IVD/GD/Stability/02/2022 Page 2 of 50
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
Table of Contents

Sr.No. Content Page No.

Preface 4
1. Introduction 5
2. Definitions 6
3. Medical Devices Rules, 2017 requirements 8
4. Basic principles for stability testing 10
5. Shelf-life studies 15
6. Component stability studies 17
7. Stability during transport 20
8. In-use stability studies 22
9. Manufacturing lots used in stability studies 23
10. Stability plan 25
11. Stability report 32
12. Changes to a Licensed / Approved IVD 33
APPENDIX
APPENDIX I Example stability protocols. 36
APPENDIX II Specimens for the stability testing 43
panels
APPENDIX III Examples of stability study 48
approaches

CDSCO/IVD/GD/Stability/02/2022 Page 3 of 50
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India

Preface:
CDSCO is pleased to announce the release of the draft Guidance Document
on stability studies of In-vitro Diagnostic Medical Device (IVDMD). It is guidance for
Manufacturers in Preparation of a Premarket Review Document for IVDMD Import
or Manufacturing License Applications.
This document is intended to aid manufacturers in the preparation of
scientific information to be provided in support of claimed shelf life, in use stability
and shipping studies for IVDMD license applications and Post approval change
application filed in pursuant to the Medical Devices Rules, 2017 (MDR-2017). This
document has been developed by the CDSCO to encourage and support
convergence of regulatory systems for medical Devices among jurisdictions.
CDSCO is looking to adopt the use of this Guidance for premarket license
applications and Post approval change applications. CDSCO strongly encourages
manufacturers to follow this guidance when submitting IVDMD license applications
and Post approval change applications.
This guidance document integrates global regulatory practices within the
Medical Devices Rules, 2017 (MDR-2017) licensing requirements for in vitro
diagnostic device license applications.
All premarket in vitro diagnostic medical device license applications are
expected to be prepared as specified in this guidance.

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India

1. Introduction
1.1. Key concepts
Stability is the ability of an In-vitro Diagnostic Medical Device (IVDMD) to
maintain its performance characteristics over a defined time interval. The
purpose of stability studies is to verify the time period and the storage
conditions over which stable performance characteristics of an IVD can be
claimed.
One important aspect of the development and manufacture of in vitro
diagnostic (IVD) medical device reagents is initially designing the stability of
a product, then determining and verifying the expiry date of the product that
is placed on the market. To determine shelf life, shipping stability, and in-
use stability, the manufacturer performs an evaluation. In order to provide
this important information to the customer, the manufacturer identifies
critical factors that might influence stability of the IVD reagent and carefully
evaluates these characteristics. Stability of the IVD reagent affects the
performance of the device and therefore has an impact on patient results.
It is the manufacturer’s responsibility to determine and monitor stability of
IVD product to ensure that performance characteristics of the product are
maintained.
1.2. Rationale of stability studies
The stability of an IVD is fundamental for its reliable performance over a
defined period of time. It is a regulatory requirement for the manufacturer to
provide objective, scientifically sound evidence to support all claims made
regarding the stability of an IVD. In addition, a manufacturer can use stability
studies to show that all lots manufactured during the commercial life of the
IVD will meet predetermined user needs.

Audience and scope

This guidance document is applicable for manufacturers in preparation of a
premarket review document for all risk-class of In-vitro Diagnostic Medical
Device for the submission of Manufacturing or Import License applications
and Post approval change applications to concerned licensing authority as
per Medical Devices Rules, 2017.

This guidance document is applicable to the stability evaluation of in vitro

diagnostic medical devices, including reagents, calibrators, control
materials, diluents, buffers and reagent kits
This guidance document is not applicable to IVD analyzers, instruments,
apparatus, equipment, software and systems.

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
2. Definitions
2.1. Definitions
The definitions given below apply to the terms used in this document. They
may have different meaning in other contexts.
• Accelerated stability evaluation:
Study designed to increase the rate of chemical and/or physical degradation,
or change, of an IVD reagent by using stress environmental conditions to
predict shelf-life.
NOTE: The design of an accelerated stability evaluation can include extreme
conditions of temperature, humidity, light or vibration.
• Acceptance criteria:
A defined set of conditions that must be met to establish the performance of
a system.
Numerical limits, ranges, or other suitable measures for acceptance of the
results of analytical procedures.
• Accuracy of measurement:
Closeness of the agreement between the result of a measurement and a true
value of the measurand.
NOTE 1: Accuracy of measurement is related to both trueness of
measurement and precision of measurement.
NOTE 2: Accuracy cannot be given a numerical value in terms of the
measurand, only descriptions such as 'sufficient' or 'insufficient' for a stated
purpose.
• Arrhenius plot:
Mathematical function that describes the approximate relationship between
the rate constant of a chemical reaction and the temperature and energy of
activation.
• Batch/Lot:
Defined amount of material that is uniform in its properties and has been
produced in one process or series of processes.
• Component:
Part of a finished, packaged and labelled IVD medical device.
NOTE: Typical IVD kit components include antibody solutions, buffer
solutions, Calibrators and/or control materials
• Constituent:
Raw materials used to make a component.
• Control material: Substance, material or article intended by its manufacturer
to be used to verify the performance characteristics of an IVD medical
device.
• Drift:
Characteristic slow change of a metrological value from a measuring
instrument.

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
• Environmental factors:
Variables that might affect the performance or efficacy of IVD reagents.
Example: temperature, airflow, humidity, light, altitude/ambient pressure,
vibration, dust, micro-organisms, etc.

• Evidence:
Information which can be proved true based on facts obtained through
observation, measurement, test or other means.

Expiry date
It is the upper limit of the time interval during which the performance
characteristics of a material stored under specified conditions can be
assured.
NOTE: Expiry dates are assigned to IVD reagents, calibrators, control
materials and other components by the manufacturer based on
experimentally determined stability properties.

• Instructions for Use (IFU):

Information supplied by the manufacturer to enable the safe and proper use
of an IVD.
NOTE: Includes the directions supplied by the manufacturer for the use,
maintenance, troubleshooting and disposal of an IVD, as well as warnings
and precautions.

In-use stability: Duration of time over which the performance of an IVD

reagent within its expiration date remains within specified limits after opening
of the container system supplied by the manufacturer and use under
standard operation conditions.

• In vitro diagnostic (IVD) medical device:

A medical device, whether used alone or in combination, intended by the
manufacturer for the in vitro examination of specimens derived from the
human or animal body solely or principally to provide information for
diagnostic, monitoring or compatibility purposes.
NOTE: IVDs include reagents, calibrators, control materials, specimen
receptacles, instrument, apparatus, appliance, material or other article
including software or an accessory used, for example, for the following test
purposes: diagnosis, aid to diagnosis, screening, monitoring, predisposition,
prognosis, prediction, determination of physiological status.

• IVD reagent:
Chemical, biological or immunological components, solutions, or
preparations intended by the manufacturer to be used as an IVD medical
device.

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
• Metrological traceability:
Property of the result of a measurement or the value of a standard whereby it
can be related to stated references, usually national or international
standards, through an unbroken chain of comparisons all having stated
uncertainties.
NOTE:- Each comparison is affected by a (reference) measurement
procedure defined in a calibration transfer protocol.
• Performance claim:
Specification of a performance characteristic of an IVD as documented in the
information supplied by the manufacturer.
NOTE: This can be based upon prospective performance studies, available
performance data or studies published in the scientific literature.
“Information supplied by the manufacturer” includes but is not limited to:
statements in the IFU, in the Device Master File supplied to CDSCO and /or
other regulatory authorities.

Real-time stability evaluation: Study designed to establish or verify the

shelf-life of the IVD reagent when exposed to the conditions specified by the
manufacturer.

Shelf-life: Period of time until the expiry date, during which an IVD reagent,
in its original packaging, maintains its stability under the storage conditions
specified by the manufacturer.

Stability: Ability of an IVD reagent to maintain its performance

characteristics within the limits specified by the manufacturer.
Stability applies to IVD reagents, calibrators and controls, when stored,
transported and used under the conditions specified by the manufacturer.
Stability includes storage and where applicable transport conditions.
Storage includes elements such as duration, temperature limits, other
environmental factors and freeze /thaw cycles.

Storage conditions – manufacturer-recommended storage conditions

stated on the product label or its instructions for use, which may include
temperature, humidity, and light protection.

3. Medical Devices Rules,2017 requirements: -

The domestic manufacturer or authorized agent (in case of Import) shall submit
the duly signed detailed information pertaining to stability of applied product in
Device Master File as specified in point 15.0 -18.0 of Appendix III of Part III of
Fourth Schedule of MDR-2017. Manufacturer should describe claimed shelf life, in
use stability and shipping stability studies and should provide information on
stability testing studies to support the claimed shelf life.

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
i. Claimed Shelf life:
This section should provide information on stability testing studies to support
the claimed shelf life. Testing should be performed on at least three different
lots manufactured under conditions that are essentially equivalent to routine
production conditions (these lots do not need to be consecutive lots).
Accelerated studies or extrapolated data from real time data are acceptable
for initial shelf life claim but need to be followed up with real time stability
studies. Such detailed information should describe:
(a) the study report (including the protocol, number of lots, acceptance
criteria and testing intervals);
(b) when accelerated studies have been performed in anticipation of the
real time studies, the method used for accelerated studies;
(c) conclusions and claimed shelf life.
Explanation- Shelf life can be derived from the lot with the longest real
time stability data as long as accelerated or extrapolated data from all
three lots are comparable.
ii. In use stability:
This section should provide information on in use stability studies for one lot
reflecting actual routine use of the device (real or simulated). This may include
open vial stability and/or, for automated instruments, on board stability. In the
case of automated instrumentation if calibration stability is claimed, supporting
data should be included. Such detailed information should describe:
(a) the study report (including the protocol, acceptance criteria and testing
intervals);
(b) conclusions and claimed in use stability.
iii. Shipping stability:
This section should provide information on shipping stability studies for one lot
to evaluate the tolerance of products to the anticipated shipping conditions.
Shipping studies can be done under real and/or simulated conditions and
should include variable shipping conditions such as extreme heat or cold.
Such information should describe:
(a) the study report (including the protocol, acceptance criteria);
(b) method used for simulated conditions;
(c) conclusion and recommended shipping conditions.

3.1. Manufacturer responsibility

It is a manufacturer’s responsibility to ensure that all claims made regarding the
stability of the IVD performance are supported by objective, scientifically-sound
evidence.

3.2. Standards
CDSCO recommends the following latest standards, guidelines for the use in
establishment of stability claims for IVD medical device:
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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
• IS/ISO 23640 (In vitro diagnostic medical devices - Evaluation of stability
of in vitro diagnostic reagents), CLSI-EP25-A, ASTM:D4169-14 and WHO
TGS2.
It is recommended that manufacturers be familiar with the standard and
consider them when designing and planning their stability studies.

3.3. Suitability for use in India

The stability studies submitted to CDSCO should accurately reflect the
expected environmental conditions and the normal usage conditions/
methods encountered by the users in India’s States, such as:
• Extremes of temperature for in-use conditions and during
transportation
• Extremes of humidity encountered during in-use conditions,
transportation and storage
• Dust
• Light, both the amount required for accurate testing/results
interpretation and any affects that light may have on the IVD
functionality
• Micro-organisms

3.4. Meeting customer requirements

By undertaking well-designed stability studies including periodic verification
activities, the manufacturer can demonstrate that the product meets
customer requirements, as required by Fifth Schedule (Quality Management
System for medical devices and in vitro diagnostic medical devices) of MDR,
2017. Meeting predetermined user expectations, not merely evaluating the
capability of an IVD, is a fundamental aspect of development of IVDs. It is a
proactive means for the manufacturer to prevent quality problems at lot
release and in the post-production and marketing phase.

4. Basic principles for stability testing of IVDs

The process to establish product stability claims follows four steps:

(1) provide an operational definition of stability for the given product;
(2) develop a stability testing protocol to generate experimental data;
(3) execute the plan; and
(4) derive and document stability claims from analysis of the experimental
data.

4.1. Critical characteristics of the IVD

A well-designed stability study must generate evidence of the stability of
each of the critical constituents of the IVD (risk-evaluated critical

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
constituents), each of the claimed analytes, and any particular level of
performance including precision, sensitivity and specificity of the kit.

Examples:
1) A hepatitis C virus (HCV) assay containing the critical constituents
related to detection of NS3 or core proteins should have the stability of all
such constituents proven...
2) For an assay designed to detect both immunoglobulin G (IgG) and
immunoglobulin M (IgM) by use of protein A and protein L, the stability of
both protein A and protein L should be proven.
3) For CD4, all the antibodies involved (e.g. anti-CD3 and anti-CD4) must
be shown to be stable.
4) For an IVD claimed to detect particular seroconversion specimens, or
genotypes, or to have specified precision at particular analyte
concentrations, or a particular specificity, each of these claims must be
proven over the stated shelf-life.

4.2. Finalized product presentation

During stability testing, all IVD components (including the device, calibrator
and / or control material, etc.) must be made and tested to the finalized
manufacturing documentation and in the finalized packaging including
intended labels and containers. All presentations (e.g. different buffer
volumes used for different kit sizes) must be used during stability testing.

4.3. Environmental conditions

The stability study should subject the IVD to a combination of conditions
which define the limits of stability for all lots made during its commercial life.
The combinations of conditions, durations of exposure and the number of
lots to be used will be driven by a manufacturer’s risk assessment for the
IVD and data from R&D. The risk assessment should take into account at
least:

• the variability of the constituent materials (identifying the most important

sources of variability);
• the nature of the users’ environments; and
• extreme conditions potentially occurring during transportation to end
users.
Boundary conditions for stability studies should reflect realistic extreme
conditions that are consistent with the design input requirements for the IVD.
The subsequent stability studies will prove the IVD capable of meeting
performance requirements at the end of its stated shelf-life, after transport to
the users.

4.4. Minimum number of lots

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
Similar to performance validation, the design of stability studies should take
into consideration lot-to-lot variation, with a risk assessment to identify the
most important sources of variability. Lot variability is caused usually more
by the biological reagents than by the actual manufacturing process.
Although existing standards recommend the use of one lot for certain
stability studies, the impact of lot-to-lot variability must be taken into
consideration and use of additional lots may be necessary. To ensure the
potential of lot-to-lot variability is addressed, optimally different batches/lots
of critical constituents such as nitrocellulose membranes, recombinant
antigens, peptides, nucleic acids and enzymes used in nucleic acid testing
(NAT), etc. that are as different as possible should be used. Refer section 3
for minimum number of lots to be considered for stability studies as per
MDR-2017.
It is advisable to test in triplicate at each point of testing interval and
maintain product, calibrator and external sample 20% extra quantities or
needs for extra during the study to accommodate all testing need.
Example: For NAT assays, it is critical to use unique enzyme lots for
stability studies. Other components including primer, probe and buffer can
also be affected by the manufacturing process (purity, pH, DNase & RNase
contamination, etc.). For these, different lots are also highly desired that
represent both material and process variability.

4.5. Assessment of liquid components:

The orientation of the product during storage (that is, upright versus inverted
or horizontal) may need to be included in a protocol. It is standard best
practice in stability studies to ensure that liquid components that are in
contact with all the parts of the container – vial, sachet or bottle, such as the
stopper, the seal and the body of the container do not affect the stability of
IVD kit. This is sometimes called “inverted container stability” but is probably
best studied by ensuring direct contact between the liquid contents of the
product and all parts of the containers by movement (orientation of IVD kit)
during the stability study. This aspect needs particular attention for in-use
stability studies of those components that are diluted or reconstituted from
freeze-dried before use.

4.6. Specimens for the stability testing panel:

The specimens used in the stability testing panels must reflect all the
performance claims related to the IVD. If a variety of specimen types (e.g.
serum, plasma, whole blood, saliva) are claimed as being suitable for use in
the instructions for use (IFU), the stability plan must be designed to provide
evidence that the IVD will maintain each of the claims (e.g. sensitivity,
specificity, proportion of valid runs, precision) for each of the specimen
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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
types for the whole of the claimed shelf-life including transport to the final
users.
Evidence should be statistically valid. The stability testing panel must be
validated accordingly and rejection and replacement criteria should be
established. Regulatory requirements may also dictate the addition of panel
members.
A stored validated stability testing panel is not always feasible. For example,
this is often the case for assays requiring fresh and/or whole blood
specimens e.g. CD4, assays to detect RNA. When replacing panel
members, the accuracy of results generated with the replacement material
must be confirmed using an appropriate reference comparator method.
Replacement criteria for unstable panel members will include the duration
for which a critical member will give valid results.
Test samples may be commercial quality controls, pooled human or animal
specimens (neat, diluted, or spiked), or other suitable materials of known
measurand stability. One sample should be at a low level, one at a high
level, and one near the medical decision level. For methods where the
medical decision level coincides with the low or high levels, only two
samples may be used.
Some IVD reagents may show little or no significant measurand drift when
tested solely with samples made from non-native analyte material (eg,
controls), yet have significant drift if tested with human or animal specimens.
As such, representatives of both native and purified or artificial sample types
should be included in method reagent stability studies to probe if the
differential recognition of the measurands changes over time. This could be
done by including at least one human or animal sample as a test sample—
ideally, with measurand content comparable to that of a control product
already chosen as a test sample.

4.7. Validation of stability testing panel

The validation of the stability testing panel members used is critical. Stability
testing panel members themselves must be stable, and they must monitor
parameters that are useful to control the characteristics being tested.
Stability testing panel members are chosen deliberately to ensure each
member has an attribute pertinent to the intended use. As with lot release
testing, the goal of stability testing is to ensure that the test method
appropriately monitors the functionality of the antigens, epitopes, and
antibodies that are relevant to the intended use at the end of the assigned
(shelf life/ in use life) life.
Example:
For instance, the intended use claim may be that early seroconversion
specimens are detected. To show that this claim is true at the end of the
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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
product’s life, a very early seroconversion specimen is included in the
stability panel. This specimen may be a weakly reactive IgM specimen.

An expected value is then assigned to each panel member and this is used
to assign the acceptance criteria for that panel member. The value for each
member is assigned in a measurable manner relevant to the outputs of the
particular methodology. For instance, the acceptance criteria for each panel
member may be assigned in terms of sample-to-cut-off ratio, cycle time (CT)
values, and band intensity measured semi-quantitatively/quantitatively.

In the example of a weakly reactive IgM seroconversion specimen, the

specimen at the start of shelf life may have a reading score on an RDT of 1+
out of 4, assigned by using a semiquantitative value based on band
intensity. The acceptance criteria may be that all reactive specimens remain
reactive, and all non-reactive specimens do not react in the assay.

As such, panel members must be chosen that not only will be relevant to
demonstrate the intended use, but have values that will appropriately detect
and therefore monitor any deleterious effects of storage. A strong positive
specimen, which has a 4+ out of 4 semi-quantitative reading value, may
remain giving this reading despite decay in the assay, whereas a specimen
with a reading of 1+ out of 4 (with an assigned acceptance criterion of
remaining positive) is more likely to give an indication of the ongoing stability
of the assay.

Thus, it is essential to know that where a panel member meets acceptance

criteria, this is a true reflection of the stability of the product and not due to
the inability of the specimen result to reflect this change.

4.8. Stability testing time intervals

A simple study design requires minimum three testing intervals:
1. an initial baseline test
2. a test at the time point beyond the claimed stability limit
3. and one point in between.

However, this is a high-risk approach that has the potential for wastage of
time and resources. If the IVD does not meet the acceptance criteria at the
end of testing, there is little information about the deterioration of the
component or IVD (or lack of deterioration) in the interim period.

A more effective approach is to test at predetermined time point intervals.

The manufacturer could identify the most practical intermediate test points

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 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
from a risk evaluation of a specific IVD. The number and length of testing
intervals should be determined in advance and form part of the stability
plan/protocol. This planning will help to understand the resources required
to execute the experiment.

Testing of all panel members is not required at all test/time points. However,
testing with all panel members is required at the initial, the second last and
the last test/time point of any of the study types. The manufacturer should
decide on practical intermediate test points at which a smaller minimal
number of panel members are tested. There should be a documented
rationale for the choice of the panels used at the intermediate test points
(e.g. representative members, specimens that are close to the medical
decision points and at the extremes of the assay range tested).

4.9. Duration of testing

Testing conducted in stability studies should extend beyond the shelf -life
determined from the user needs. The shelf-life should be assigned based on
a risk assessment of the lot-to-lot variability in signal change at the end of
shelf life. At a minimum, testing should extend at least one time point (one
testing interval) beyond the determined user requirement. This provides a
safeguard in the event of unexpected IVD failure at the end of the testing
period, in which event extrapolation from an earlier time point would not be
considered acceptable.

It is recommended to utilize standardized units of measure for the entire

study.
(e.g. Unopened kit shelf life are always measured in months; opened kit
/reagent stability in days or weeks)

5. Shelf-life studies

5.1. Requirements for determination of shelf life

The stated shelf-life of an IVD must be based on real-time experimental results.
Accelerated studies or extrapolated data from real time data are acceptable for
initial shelf life claim but need to be followed up with real time stability studies.
Such detailed information should describe:

(a) the study report (including the protocol, number of lots, acceptance
criteria and testing intervals);

(b) when accelerated studies have been performed in anticipation of the real
time studies, the method used for accelerated studies;

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Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India

(c) conclusions and claimed shelf life.

Explanation: Shelf life can be derived from the lot with the longest real time
stability data as long as accelerated or extrapolated data from all three lots
are comparable.
5.1.1. Real-time stability studies

Real-time stability is determined using storage temperatures derived from user

requirements, over a period longer than the required life of the IVD. Where a
broad range of storage temperature is claimed by the manufacturer (e.g. “Store
at 4–40°C”), CDSCO expects the studies will provide evidence for stability over
the whole of the temperature range (Min, Max and in between) for at least the
length of the claimed shelf-life. Exceptionally, where claimed stability is
restricted to a limited range (e.g. “Store at 2-8°C”), it is acceptable that stability
studies are conducted at a single temperature within this range.

A sequential approach should be used, in which IVDs are first submitted to

stresses simulating transport before they are placed into a shelf -life or in-use
study. This approach best simulates the real-life situation, where products will
first be transported to the end-user and then stored under the recommended
conditions before use, possibly almost until the end of their labelled shelf -life.

5.1.2. Accelerated stability studies

Accelerated stability studies are designed to predict the shelf -life of an IVD from
the increased rates of chemical and/or physical degradation caused by extreme
environmental conditions (e.g. elevated temperature at higher humidity).

If the Arrhenius equation or other suitable data analysis is used to calculate the
expected life at temperatures other than those actually used, then the
parameters of the equation must be derived from the experimental data and not
assumed.

Accelerated stability studies provide results in a relatively short time. However,

the results of these studies are made using assumptions about the degradation
of reagents and IVD components that may not reflect their performance under
normal conditions of storage and use.

5.1.2.1 Considerations for Planning Temperature-Based Accelerated

Stability Studies
Planning accelerated tests involves decisions on the number of elevated
temperature levels, number of replicates, and number of time points for each
temperature and duration of testing at each level.
For example: if an IVD is stored under recommended storage conditions of 2 °C
to 8 °C, then accelerated stability studies might be run using product stored
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Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
typically at three or five elevated temperatures (15 °C, 20 °C, 25 °C, 30 °C, and
45 °C), taking care to avoid denaturing conditions.

6. Component stability studies

6.1. General principles

6.1.1. Testing on final specifications
Component stability studies, including antimicrobial and desiccant studies,
must be performed using components made according to finalized and
approved manufacturing specifications – ideally to validated
manufacturing scale – on qualified manufacturing equipment and meeting
finalized and approved in- process quality control (QC) specifications.

6.1.2. Considering component stability

Sometimes components of IVDs are prepared in bulk and stored before
being used in several different lots of a completed IVD. The design input
documentation should define how long components are likely to be stored
before use. With that information, component stability studies should be
planned to give evidence that component shelf lives will not restrict IVD
shelf life, since an IVD cannot have a shelf life beyond that of any of its
dependent components.

Shelf-lives of components manufactured in bulk and used in several

different lots of an IVD can be verified as for the IVD itself – three lots of
the component as a minimum for shelf-life studies and, depending on
documented risk assessment related to variability, one or more lots
subsequent to change. The evaluated lots of the component must differ in
batches of critical constituents but, again subject to documented risk
assessment, may all be tested in their final presentation with a single set
of the other components which will be used together to constitute the IVD.

Examples of stored components:

Wash solutions and substrates for enzyme immunoassay (EIA),

amplification reagents for nucleic acid testing, calibrators for quantitative
tests, manufactured and stored in their final labelled vials ready to be put
into a kit. Component stability can be assessed from the functionality of
the lot and also by factors related to the component itself, such as
turbidity, colour change, microbial contamination and pH of liquid
components changes over time.

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Ministry of Health and Family Welfare, Government of India
Depending on the IVD and the conditions it is subjected to it may be
necessary to distinguish between turbidity that arises from heat/cold
denaturation and turbidity that arises from microbial contamination.

6.1.3. Considering constituent stability

The plan should also consider whether components made from new
constituents (antigens, recombinant antigens, enzymes, antibodies,
membranes) will have the same lives as components made from stored
raw materials. Although this aspect is difficult to study, some evidence
should be provided supporting the use of stored constituents, as well as a
plan to evaluate lot-to-lot variance from different critical constituents.

The choice of the reagents to be used to measure the performance of the

constituent under study (either materials of proven shelf -life or freshly
made) needs substantial consideration.

Examples of stored constituent: Purified recombinant antigens and

monoclonal antibodies stored in aliquots ready for dilution and striping
onto RDT membranes or other supports.

6.2. Stability of control materials

Assay specific control materials provided by the manufacturer are to show
that an IVD has performed as intended during use. The manufacturer must
be able to demonstrate that the loss of signal from a control material does
not occur at a different rate from the loss of signal from a validated stability
testing panel member or from genuine, critical specimens; otherwise a failed
IVD might be regarded as still functional. Thus the stability of the control
material must accurately reflect the stability of the assay. A control that is
more stable than the IVD and other components, or incorrectly set values for
the control material, must be avoided.

Example: It is seen in Device Master Files relating to IVDs submitted by the

manufacturer that a positive run control will produce a signal of >2.0 optical
density (OD) in a freshly manufactured lot, and the IFU will state that an OD
> 0.8 for the same control qualifies a run. Thus, the IVD could have lost
more than half its activity and still appear functional, although some critical
specimens are shown in the Device Master File to have very weak signals
on freshly made IVDs. This is not considered appropriate unless data can
be provided that demonstrate that the critical specimens will still be detected
at the end of shelf life.

6.3. Antimicrobial stability and efficacy

6.3.1. Rationale

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IVDs are used in areas which are not necessarily clean and sterile, and
antimicrobial preservative may not effective under some circumstances.
Bacterial and fungal organisms relevant to the environment of use should be
identified in the design input risk assessment, and antimicrobial preservatives
should be chosen to avoid contamination of the product. The manufacturer must
obtain evidence that the antimicrobial preservative and concentration chosen is
stable and effective against the micro-organisms of concern throughout the
claimed shelf-life and in- use shelf life of the IVD

6.3.2. Study conditions

The studies should reflect expected in-use conditions in opened containers:
clean, particle-free laboratories do not usually reflect universal user environment
for suggested methods. Examples of bacterial groups to consider are spore-
forming bacteria, fungus, indigenous bacteria, bacteria found in the environment
of the country of manufacture, as well as use of a negative control. Specific
examples include Aspergillus niger, Bacillus subtilis, Candida albicans,
Escherichia coli, Salmonella species, Pseudomonas aeruginosa, Clostridium
sporogenes and Staphylococcus aureus.

Antimicrobial preservative effectiveness, as measured by the viable microbial

species load present in kit components, should be demonstrated during
development, during scale-up, and throughout the shelf-life.

Testing for antimicrobial preservative content should normally be performed at

release. Under certain circumstances, in-process testing may suffice in lieu of
release testing. The acceptance criteria for in-process testing should remain part
of the specification.

6.4. Desiccant functionality

Desiccants affect the stability of the entire IVD. Stability studies must show that
the desiccant will support the product over the whole claimed shelf -life within the
predetermined extremes of transport, storage and in-use conditions.

Note:
1) CDSCO recommends that a self-indicator (a humidity indicator that changes
colour upon saturation) be part of the desiccant design. However, CDSCO
strongly recommends against the use of cobalt dichloride, the most
commonly used humidity indicator, as it is a carcinogenic substance.
2) Sachets are preferable over tablets, since labelling such as “Do not eat” is
more visible. There have been anecdotal reports of desiccants in a tablet
formulation being mistaken for antimalarial medicine.

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7. Stability during transport

7.1. Rationale
Transport/ Shipping stability studies evaluate the tolerance of an IVD to the
kinds of environmental conditions (e.g. temperature, humidity, dust) and physical
conditions (inversion, vibration, physical handling, stacking) to which it is likely to
be subjected in the time between shipping from the manufacturer to its final
user. They should provide evidence that there will be no impact on the IVD
performance over the whole of its stated shelf-life after recommended
transportation methods for the IVD. The manufacturer should assess the
potential impact of multiple factors and justify and document whether or not to
include them in the evaluation.

CDSCO expects that a transportation challenge should precede the real-time

determination of shelf-life and in use studies. This serves to determine that
transportation conditions do not reduce the shelf-life of the IVD.

In some cases, it might be acceptable to test the product only over the transport
simulation duration, without a subsequent long-term study under normal storage
conditions. If that is done, shelf-life must be established under specified storage
conditions along with a stringent, evidence-based risk assessment of the
probabilities of extreme transport stress affecting the performance at the end of
the claimed life.

7.2. Challenge conditions

Determination of the stability during transportation of an IVD should take into
consideration the local routes, transport means and transit used to supply the
IVD, usually defined in the design input risk assessment. It is not necessary to
test the IVDs to the point where it is no longer usable, but merely to validate the
window of transport conditions within which the IVD will retain its claimed
performance to the end of its stated shelf-life. However, knowledge of the
possible limitations of an IVD and at what point the IVD becomes unusable is
useful to a manufacturer when trouble shooting post-market problems. CDSCO
expects the manufacturer to consider that the product might continue to be
subjected to sub-optimal storage conditions at the end-user.

Example: While a static challenge of 45°C for 3 days might represent conditions
seen during actual transport of a IVD, a more stringent challenge of cyclical
higher and low temperatures (including freezing) for a longer period of time and
under vibration might better cover a ‘worst case scenario’ of shipment, storage
and subsequent transportation to the end-user.

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7.3. Number of lots
For transport/shipping stability studies, at least one lot of the IVD can be
used.

7.4. Multiple stress test sequences

Appropriate sequences may be developed on the basis of data from actual
product transport studies. Testing multiple stress sequences allow a
manufacturer to identify the most cost- and/or resource effective transport
conditions from a set of alternatives while ensuring adequate product
stability protection.

Note: Environmental conditions investigated as part of a stability study must

reflect those likely to be encountered in resource- limited States.
Temperatures at some airport tarmacs can exceed 40°C while temperatures
encountered during air transport fall below 0°C. Significant delays can be
encountered at all times and especially during wet season transport to
remote health centres.

7.5. Physical conditions

Physical handling can be both manual and mechanical. The relevant user
and commercial factors should be identified as part of the design input risk
assessment and the packaging and shipping methods developed
accordingly. Reference defines a number of factors to be considered, and
their evaluation in the shipping stability protocol: drop, impact, compression,
vibration, repetitive shock, longitudinal shock, cyclic exposure, vacuum,
impact, inversion; along with the size, weight, and composition of the
packaging.

7.6. Simulated versus actual challenge

An actual shipping challenge can be used to verify the conditions found in
the simulated transportation challenges. However, it should only replace a
simulated shipping challenge when there is an appropriate risk evaluation
and with experience and data already actively collected from similar
products and documented in detail (for example it is insufficient to note “no
complaints”). In the R&D phase, actual data from shipping can be used to
define the conditions needed for an appropriate simulation of extremes.
However, in the post-production phase actual shipping challenges often do
not explore the full range of shipping conditions that could be encountered,
including extreme values.

8. In-use stability studies

8.1. Rationale
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In-use stability of an IVD is the period of time over which components retain
adequate performance, after transport to the users, once they are opened,
reconstituted and/or diluted and exposed to the environmental conditions in
which they will be used.

If a range of conditions for use is stated in the IFU (e.g. use at 15–40°C)
evidence should be provided to prove the stability over that range with all
the specimen types (e.g. serum, whole blood, oral fluid) claimed. It is
considered best practice that the manufacturer extends the stability range
by 5°C at the lower and upper end of the proposed acceptable range on the
labelling for all components to ensure that that the claimed stability ranges is
acceptable.

8.2. Conditions of use

Determination of the in-use stability of an IVD and/or its components should
reflect recommended conditions of use of the IVD. Freeze-thaw stability
should be considered to address reagents which are exposed to multiple
freeze-thaw during use.

Note: In-use stability studies must take into account environmental

conditions and usage conditions encountered by users and States, such as
exposure to extreme temperatures, humidity, dust, light and micro-
organisms.

8.3. Multiple in-use stability claims

Depending on the way in which the IVD is used it may be necessary to have
several in-use stability claims. In situations where multiple stability claims
are made, a manufacturer must provide evidence from testing that
investigates routine use supporting each of the claims.
Examples:
1) A reagent may have a stated period of stability once it has been placed
on-board an instrument and another period of stability once it is in active use
(i.e. during actual use/testing).
2) Multiple use reagents (e.g. buffers) may repeatedly be exposed to high
temperatures during the day while in use and exposed to lower
temperatures when not in use and stored in the refrigerator. The actual use
of the multiple use reagent – squeezing of bottles, exposure of the lid and tip
to working surfaces, hands, exposure to dust and light – also affect stability.
Stability studies should take into account all of these factors.

9. Manufacturing lots used in stability studies

9.1. Considering variability
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Stability studies must take into consideration all possible sources of
variation within and between manufactured lots. For most IVDs it is likely
that differences between batches of the biological reagents will cause the
most variance. Factors to consider include apparently minor, technically-
uncontrollable differences in culture and purification for recombinant
antigens and antibodies; synthesis and purification for primers, probes and
peptides; undocumented production changes of an outsourced buffer
component and the lot of nitrocellulose membrane used in lateral-flow IVDs.

At a minimum, lots chosen for stability studies should be different in the

critical constituents, e.g. different purification and/or culture batches for all
recombinant antigens and monoclonal antibodies. If pilot or small scale lots
are chosen, special attention must be paid to the potential for variability.
However, the sources of variation will depend on the particular process,
product and component, and should be identified during product
development risk analyses.

Use of different batches of critical components ensures that the stability

evidence obtained is more likely to be representative of long-term
manufacture. Any variability found can be taken into consideration when
assessing the outcome of the studies against the design input requirements
and when making claims. This minimizes user problems and hence
complaints.

9.2. Testing the final configuration

Shelf-life, in-use and shipping stability must be determined for the finalized
product configuration, in terms of:
• manufacturing specifications;
• release-to-market QA criteria;
• packaging and labelling; and
• validated manufacturing scale on qualified manufacturing equipment.

Note:
Testing methods should be as included in the IFU of the finalized IVD.
It is important that it can be established that the stability studies were
conducted on the IVD as submitted to CDSCO for approval. Even changes
perceived as small (e.g. change in production scale, bulk container
materials, supplier of a critical biological, change in vial stopper) can have
unexpected effects on stability and other performance characteristics. After

such changes, a stability plan and study is needed again. Manufacturers

should have change control procedures in place compliant with ISO 13485.
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Stability studies undertaken in the R&D phase of the product lifecycle are
important to understand how to design the product so it will meet the final
stability requirements in the input documentation. However, these studies
are not sufficient for submission to CDSCO since they might not reflect the
final design and manufacture of the IVD.

9.2.1. Exceptions
If any of the above criteria are not met (for example if “pilot lots” or small
scale lots are used, or if the IFU is not finalized), strong evidence must be
provided that the evaluated materials will perform exactly the same as the
final product.

Note: In some exceptional circumstances, where it is not possible to sample

from actual production lots, samples from pre-production or development
lots might be used. If this is the case, manufacturers should justify why
production lots were not used, and they should provide robust evidence that
the lots chosen are expected to behave identically to the production lots.
Data concerning lot-to-lot variability must still be submitted. Although
CDSCO will consider the available evidence on its merits, this preliminary
information must be followed by stability claims conducted on production
lots. A post-approval commitment may be required to amend this situation
when the manufacturer is able to produce fully qualified production lots.

9.3. Number of lots required for testing

As per MDR-2017, accelerated and real time stability study testing should be
performed on at least three different lots manufactured under conditions that
are essentially equivalent to routine production conditions (these lots do not
need to be consecutive lots) be used to verify shelf -life; one lot reflecting
actual routine use of the device (real or simulated) be used to verify in-use
claims; one lot to evaluate the tolerance of products to the anticipated
shipping conditions be used to verify Shipping stability.

Note:
It is not acceptable to sample IVDs from a single manufactured lot but label
them so that they appear to have been taken from three separately
manufactured production lots. This aspect will be investigated during an
onsite inspection by CDSCO. Non-compliance with this requirement may
result in a major Non-compliance under the MDR 2017.

9.4. Components of lots required for testing

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Stability work is performed using materials in their final packaging, with
intended labelling. If there is more than one variant of the IVD (e.g. pack
size differences) any potential effects on performance, including stability
must be assessed.

In particular, if different reagent-container sizes are used in packs intended

for different numbers of use, stability evidence should be obtained on all
variants, even if the contents of the containers are identical.

Once component shelf-lives are assigned, use relatively fresh components

and components which have progressed into their assigned shelf -life in the
different production lots used in the establishment of the product shelf -life.

Labelling is a significant factor of packaging and may affect stability in some

cases. For example, some label adhesives diffuse through some plastics,
enter vials and affect the function of the reagents over time. Other label
types lose adhesion over time; while some printing inks fade. The physical
stability of packaging requires the same degree of risk evaluation and
subsequent experimental verification as its chemical stability, with attention
given to the countries of intended use. This is most important for primary
packaging but must also be considered for secondary packaging,
particularly for transport stability studies.

10. Stability study plan

Stability studies should be well designed, scientifically sound, well

implemented, well recorded and able to deliver meaningful conclusions about
IVD performance.

This will minimize the time and resources taken by the manufacturer to
generate appropriate evidence and by the regulatory authority to assess it.

It is good practice to prepare, within the mechanisms of a Quality

Management System (QMS), a plan for the investigation of each aspect of IVD
stability. A well-developed study plan, with clearly defined objectives,
responsibilities and pass/fail criteria should be developed, reviewed and
internally approved in advance of testing. The protocol should be associated
with the design input requirements.

It is essential that the study protocol takes into account the intended use of the
product to ensure that these elements are covered within the stability studies.
The results of the stability studies support the claims in the instructions for
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use. Careful forward planning will make a significant contribution to ensuring
that sufficient resources are made available, effective experiments are
performed and both experimental results and associated documentation are
recorded in an appropriate manner.

10.1. Responsibilities
The study protocol should outline responsibilities and applicable training for
said responsibilities of all staff involved in the study.

The responsibilities for implementing the study plan must be assigned to

appropriately qualified and trained staff. Responsibilities to be allocated
include study set up, testing, monitoring, validation of equipment and/or
processes, sample selection, risk assessment and corresponding
documentation.

In addition, the manufacturer must nominate a person responsible for

investigating failures and a person responsible for conducting risk
assessments if the IVD fails to meet the requirements of the design inputs.

10.2. Preparing the testing plan

A complete, detailed description should be prepared that fully documents
everything to be done and the expected outcomes. Authorization of the
protocol should be obtained internally in advance of starting work. The
protocol should include the following details.

• Qualification and training of technical staff performing the work

• Biohazard issues identified with reagents
• The instrumentation, including storage facilities or rooms, validation,
calibration, monitoring, servicing
• The batch/lot numbers of kits to be used with justification for any
manufacturing anomalies or deviation from documented procedures
• The expected life of the kit from the input documentation
• Any proposal, with justification to launch a kit with a life based on
accelerated data, or to launch with a shorter life than in the input
documentation while awaiting the conclusion of real-time testing
documented.
• The documented nature and extent of in-use testing
• The justification for the choice of lots and components taking into account
lot-to-lot variation and the critical characteristics
• The number of units (cassettes, bottles, tablets, etc.) of each component
to be collected and stored under each condition

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• The nature of the stability testing panel to be used, justifying each panel
member’s inclusion and defining the volume and characterization of the
bulk specimen to be used and the aliquot size and number to be stored
for the testing
• The expected criteria for each stability testing panel member at the
beginning and end of the product’s proposed shelf-life
• The statistical methods to be used for data analysis
• Graphs (paper or electronic) to visualize the performance of each stability
testing panel member over the course of testing
• Methods of approval and justification of any deviations from the plan

10.3. Product storage

A sufficient number of product components from the identified lots should be
reserved and stored separately to ensure that the study will be completed
with identified products. Sufficient number of IVDs should be retained to
allow for additional testing, calculated from the predetermined invalid result
rate.

10.4. Documentation
The Stability protocol should make reference to a study report which will be
used to summarize interim, and ultimately, final study findings and
conclusions. The study plan, the testing protocol and study report and all
associated documentation (worksheets, etc.) should be controlled within the
manufacturer’s QMS. At the end of the study, the manufacturer should be
able to confirm that design input requirements have been met.

Any changes in method must be recorded and undergo risk assessment. It

should refer to the development of a detailed and valid testing protocol
which includes all information and material relevant to testing.

10.5. Statistical methods

Statistical methods are used to support stability claims by providing
estimates of the probability of results being as stated. For example: prior to
the stability studies on an EIA it has been documented that if a stability
testing panel member has at least a particular optical density (OD) then that
device will meet a particular claim. Given the results of the stability study

using that stability testing panel member and showing the variability within
and between lots of the IVD, the probability of future similar production of
the device meeting claims at the assigned life can be estimated. The
derivation of valid criteria and the probability of maintenance of all claims
can be estimated by appropriate statistical methods.
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A fundamental problem is that of how many replicates should be used at

each time point and from how many different production lots to produce
acceptable overall probability estimates of the likelihood of all future
production of similar devices and lots meeting claims (and hence user input
requirements) at the end of the assigned life. There are two aspects to this –
what is “acceptable” and “how many replicates?” “Acceptability” is a decision
critical to quality and must be decided in advance from the user
requirements – for example 80% confidence that 95% of all lots will meet
the claims. This is in fact a tolerance interval as described in ISO 16269-6:.
“How many replicates” can then be derived from the tolerance interval
required but advice from a professional statistician is strongly advised –
after defining the quality critical requirement but before beginning any
experimental work.

The statistical methods to be used will be documented in the plans and

protocols of any stability study and consideration is given to treatment of
unexpected and atypical results. In general, all results must be used unless
there is a documented physical reason (e.g. known operator error, too little
volume, incorrect timing, use of an unqualified instrument such as lacking
maintenance or calibration) why a result can be ignored – but even then that
result must be recorded and included in the report of the stability work.

10.6. Stability testing protocol

As part of an approved study plan for the determination of IVD stability, a
detailed testing protocol should be prepared (examples of stability protocols
are provided in Appendix 1: Example stability protocols) including the
following as a minimum, as appropriate.

• QMS identifiers (e.g. experiment name, document references, etc.)

that allow traceability to both the overarching study plan and to
subsequently generated records/documents such as result
worksheets
• Responsibilities, name(s) of operator(s) and their training
requirements.
• The dates and times when the stability study will be performed
• Name, designation and Signatures of the operator and supervisor(s)
• The objectives of the study (i.e. determination of shelf-life,
determination of in-use stability of a component, etc.)
• The name and lot number of the IVD and/or components being
subjected for stability studies.

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• How the components/finished product will be sampled from the
production department
• Stability testing panel members and their characterization to be used,
including valid test methods which reflect the IFU claims.
Note: IVDs should be tested with samples at several different analyte
concentrations, including samples at low concentration near the cut-
off level of the assay.
• The experimental method that will be used for testing. This must
follow the finalized testing method from the IFU. It must describe
clearly how the experiment was performed in terms of:
➢ required storage and/or challenge conditions;
➢ the duration of storage/challenge;
➢ the schedule of testing intervals;
➢ the stability testing panel; and
➢ the numbers of replicate tests performed for each stability testing
panel member.
• How and where results are to be recorded
• Acceptance criteria
• How aberrant, discordant or invalid results will be dealt with
• How storage/challenge conditions are to be applied
Example: For determination of stability during transportation it should
be made clear that each IVD will be subjected to a sequence of stated
temperatures.
• How actual storage/challenge conditions are recorded
Example: Recording of temperature not as “room temperature” but as
an actual numerical value obtained from calibrated instrumentation

Note: It can be unclear to a CDSCO reviewer from a general statement

such as “… Sample buffer was stored at the required temperature and
tested each month…”. This statement raised questions such as whether (1)
the bottles of sample buffer were stored open at the required temperature
for the entire testing period, or (2) the bottles were stored capped and
refrigerated, and only reopened briefly at the required temperature at each
schedule test point. To avoid confusion, the details of actual storage and
use procedures are required in the testing report.

10.7. Reading and recording results

10.7.1. Avoiding reader bias

It is good practice to use approaches to make the reading more objective,
such as a scoring system. For IVDs where a subjective element forms part
of the result, e.g. reading the intensity of an RDT band within a specified
time frame, the results should always be reviewed by a first and second
reader to avoid operator bias. Both readers must be blinded to the
expected results; the second reader must be blinded to the first reader’s
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results. If a validated band intensity scoring tool is to be included in the
final RDT kit, this should be used to record results.

10.7.2. Recording actual individual results

The results of a test, not only the test interpretation, should be recorded.
An interpretation on its own has insufficient resolving power to allow
degradation of a signal over time to be observed. Some IVDs, e.g. line-
blots, may require particular band patterns to allow an interpretation to be
reached, and several different patterns may yield the same final result.
Recording only the final interpretation of a test specimen may cause the
failure of particular bands to go unnoticed while allowing the IVD to
otherwise “pass”. Photographic records of qualitative tests are
recommended, as appropriate.
This is particularly important when testing a panel of like specimens, e.g.
“20 HIV antibody positive specimens” for which the acceptance criterion is
“all 20 specimens must be positive”. It is not sufficient to simply record “all
20 positive” or “pass” without first recording the individual test result
directly from the IVD for each specimen in the panel.
Example 1: For most enzyme-linked immunoassays (EIAs) if the sample-
to-cut-off ratio is > 1 then the result is interpreted as “positive” or
“reactive”. In this case three pieces of information should be recorded: (1)
the numerical value of the assay sample-to-cut-off ratio, (2) the numerical
value of the signal for the specimen and (3) the final interpretation.
Example 2: Some rapid diagnostic tests (RDTs) may stipulate that the
strength of test band is not correlated with the strength of antibody titre.
Nevertheless, the following should be recorded: (1) the intensity of
observed patterns according to a predetermined, validated intensity
scoring system with as fine a gradation as possible, and (2) the final result
interpretation.
Example 3: A qualitative NAT assay may report “positive” and “negative”
for a particular analyte, but the underlying decisional parameter is often
quantitative (e.g., a PCR signal-based cycle number). The quantitative
parameter should be recorded.

10.7.3. Retention of records

CDSCO encourages retention of photographic records, machine printouts
(in case of thermal printout, a photocopy of the same duly signed by the
operator needs to be kept), electronic data or physical retention of
membranes from opened cassettes, as appropriate. The manufacturer
shall retain records for a period of time at least one year after the date of

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expiry of IVD as defined by the manufacturer, but not less than two years
from the date of product release by the manufacturer.

Instability versus imprecision: Testing at more than two time points can
be important to avoid confusion between imprecision and instability. For
example, if the end testing shows 10% decrease, one may not judge if the
difference was due to imprecision or instability. The inclusion of additional
test points (for example one or more between the zero time and the end of
the shelf-life) allows for fluctuation caused by imprecision to be
distinguished from drift due to instability. This can be gained by increasing
the number of replicates and runs. All studies should support precisely
defined time periods of in-use stability claims.
Example: An RDT test cassette – may be labelled “Use immediately on
opening”. In such cases it is still necessary to determine the interval (one
hour, one day, etc.) over which the IVD performance remains stable after
the component is opened.

10.8. Testing schedule

Testing intervals should be selected to detect any trending of results over
the testing period. Concurrent testing of separate types of components may
be approached with different intervals. For example, it may be appropriate to
test an IVD test cassette against a stability testing panel on a monthly or
quarterly basis, but to test for open vial stability on a weekly basis.

10.8.1. Acceptance criteria for results

The acceptance criteria to establish what is acceptable or not acceptable
should be defined according to the stability testing panel criteria for both
qualitative and quantitative test methods.
In order to determine the duration over which a product can be deemed
stable, it is essential to establish the maximum allowable change for each
stability metric before starting a study. These acceptance criteria may
come from the product’s design input requirements, especially its intended
use, or other quality specifications, historical data of similar products, or
typical performance of an existing product (for the stability qualification of
product design changes).

Results from failed (invalid) test runs must not be used in the
determination of the stability claim. However, the invalid results should
also be recorded.

11. Stability study report

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11.1. General
After testing has been completed, the findings should be summarized in a
stability study report. The report should clearly identify the IVD that was
tested, date of test carried out, the objectives of the study, the conditions
under which the IVD was tested and conclusions that were drawn from
findings. The report should be traceable to the study plan, testing protocol and
user needs. It should make clear references to other supporting
documentation (e.g. result worksheets).

11.2. Link to claims

The results and conclusions of stability studies presented in the study report
must support the claims of IVD stability reported in the IFU and elsewhere in
the Device Master File.

11.3. Consider variability

An overall stability claim (whether for shelf-life, in-use stability, or shipping
stability) must be based on the expected stability when taking into account
inter-lot variability.
Example: The manufacturer should evaluate the variability between the
different lots studied and assume that any differences in shelf -life are
inherent to the manufacturing process. The claimed life should be calculated
so that a known and stated proportion of all lots (usually >95%) will meet the
claimed shelf-life. Frequently more than three lots are needed to obtain a
realistic idea of the variability of the results.

11.4. IVD stability versus component stability

A claim of stability for an IVD as a whole must not exceed any individual
component stability.
Example: For an IVD claimed to detect HIV-1 and HIV-2 antibodies – if
detection of HIV-1 antibodies is stable to 24 months but that of HIV-2 to only
18 months, then the shelf-life must be based on the shorter time i.e. 18
months.

12. Changes to a Licensed / Approved IVD

Any major modification to a licensed / approved IVD or to its process of

manufacturing will require provision of direct evidence of stability. An
appropriate risk analysis and an accelerated stability study comparing the
original (licensed/approved) product and the modified product for usability,
performance and lot-to-lot variation may serve to assess the impact of the
changes to a product formulation or manufacture. It would be necessary to
validate the stability of the modified IVD in at least one lot of the IVD (subject

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to risk analysis) in order to demonstrate equivalence between the original and
modified IVDs. More lots may be appropriate depending on the product
nature, variability of components and failure risk. CDSCO expects results of
accelerated testing to be confirmed by real-time studies.

The license holder shall obtain prior approval from the concerned Licensing
Authority, before any major change as specified in Part A of Sixth Schedule of
MDR-2017 is carried out. If there are different presentations, the stability of
each one must be assured.

The following examples seek to illustrate the scope for considering the
performance evidence from one IVD as support for performance in another:

Examples:
1) For an HIV RDT which uses an identical cassette and physical components
of a manufacturer’s existing, fully validated HCV RDT, the reagent
formulations are different (antigen/antibodies, buffers, conjugates, etc.).
Evidence of stability of the HCV RDT would not suffice for the HIV RDT. Even
if the manufacturer claims that both IVDs have been sold in a number of
countries for several years and no adverse feedback has been reported, this
would not constitute evidence in support of the stability of either IVD.

2) For an HIV RDT that has been fully validated for detection of HIV-1
antibodies; a new product is developed which includes detection of HIV-2
antibodies. The stability of any sample buffers that are identical between the
two IVDs would probably not need to be validated. However, other
components (conjugates, antigens, antibodies) that are different between the
two IVDs would need to be tested; it would not be sufficient to assume that
HIV-1 reagents will have the same stability in the new IVD. A modification of
this nature is likely to require substantial validation of stability.

3) An HIV RDT IVD previously intended for testing serum/plasma has added
to it a claim for detection of HIV-1 in whole blood. The only substantive design
change associated with the new claim is the addition of a small pad of some
suitable material near the sample port which acts as filter for whole blood
specimens.
Depending on the nature of the material it may be reasonable to argue that the
material would not be expected to age; that it is not, in any practical sense,
chemically labile. Consequently, shelf-life and in-use stability may not
necessarily need to be retested in full. However, stability during transportation
may need to be determined to provide confidence that the modification is able

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to withstand likely shipping conditions (e.g. that the extra square of filter paper
doesn’t dislodge when packages are jostled and bumped in transit).

4) Based on an HIV RDT that has been fully validated for detection of HIV-1
antibodies, a new IVD is developed which includes detection of antibodies to
Treponema pallidum (TP). Detection of TP specific antibodies occurs on a
completely separate membrane (and associated architecture) to that of HIV
antibody detection. Additional handling steps may have an impact on the
stability of the HIV-1 antibodies and it may be required to retest. It may be
necessary to review evidence of stability during transportation to ensure that
new components are not affected by transport (for example a new packaging
concept is used).
• If a new machine is used for striping of the HIV-1/TP IVD, validation of the
new machine (installation qualification, operational qualification and
performance qualification) would be required to show that the stability
studies are still valid.
• If the IVD is designed in a way that HIV and TP detection occurs either on
the same membrane and/or using most of the same architecture (and
assuming that sample buffers are identical between IVDs) it is likely that
this new IVD would need to be fully validated.
It should be noted that these observations pertain specifically to IVD stability.
Other aspects of IVD performance should still be validated as appropriate.

12.1. Change in the Approved Shelf life:

If any change with respect to approved shelf life (extension or reduction of shelf
life) shall be consider as major change as per Sixth Schedule of MDR 2017, the
licence holder shall obtain prior approval from the CLA (Central Licensing
Authority) or SLA (State Licensing Authority) as the case may be by making
application through the Medical Device Online portal. Further, during premarket
approval product may got approval as per available real time stability data later,
under Post approval change (PAC) may get extended with supporting real time
stability data.

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References:

1. ISO 23640:2011. In vitro diagnostic medical devices – Evaluation of

stability of in vitro diagnostic reagents. First edition 2011-12-01.
2. Evaluation of stability of in vitro diagnostic reagents; approved Guideline
EP25-A. Wayne (PA): Clinical and Laboratory Standards Institute;
September 2009.
3. Technical Guidance Series (TGS) for WHO Prequalification – Diagnostic
Assessment: Establishing stability of in vitro diagnostic medical devices
i.e., WHO/BS/2017.2304 Rev. 1

APPENDIX
Appendix I Example of stability protocols
Appendix II Suggested specimens for stability testing panels
Appendix III Examples of Stability Study approaches

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APPENDIX – I

Example of stability protocols:

This appendix contains examples for a wholly fictitious IVD, illustrating the kinds of
experimental design to determine the following:
1. Stability of whole kit during transport
2. Stability of whole kits during shelf-life, and
3. In-use stability of whole kits including reagents
The information provided in these examples should not be taken as a checklist of
sufficient conditions, but manufacturers are encouraged to use as a guide on
possible approaches to generate evidence of a standard sufficient to satisfy the
requirements of the MDR-2017. It is recommended that shipping/ transport stress
studies are undertaken prior to the shelf-life studies and in-use studies.

Description of fictitious IVD

IVD used for the purpose of the examples is a RDT for the detection of antibodies to
HIV-1, HIV-2 and Treponema pallidum in serum, plasma and whole blood. It is
recommended that the kit is stored at 8–40°C, but components of the kit must be
used at 15–30°C. The product is supplied as a kit with each test cassette sealed in
a foil pouch (with desiccant). The pouch must be brought to 15– 30°C. Once
opened, it is recommended that the cassette is used immediately. The IVD includes
a bottle of specimen buffer/diluent for use with all three specimen types. The
specimen buffer is expected to have similar stability as the test cassette in its
unopened form. The stability of the opened bottle of specimen buffer is determined
below (see Example 3: In-use stability protocol).

The manufacturer of this product proposes to determine the stability of its product
and has written a stability plan. As part of this plan a preliminary determination of
accelerated stability has been conducted at several extremes of temperature and
suggests that the IVD would be stable to an equivalent of 12 months following
manufacture. The plan now calls for the development of real-time stability protocols
that will form the basis of subsequent testing of the IVD.

Preliminary work has shown that the variability between lots is minimal so that three
independent lots (no critical constituents in common) will suffice to enable a
reasonable estimation of shelf-life taking lot variation into account.

Example 1: Evaluation of shipping stability

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Objective:

To determine the stability after transportation of the HIV RDT in real-time using
simulated shipping conditions and to generate stressed components to be used in
real-time shelf-life studies as proposed in Stability Study Plan xxxxx01.

Preparation

Acquire sufficient numbers of IVD kits from three independent production lots using
a predetermined sampling protocol (e.g. random, first X number of kits in first box,
every 100th kit, etc.). Allow at least 10% for unexpected requirements and re-testing

Note 1: To provide security against unforeseen events, duplicate tests should be

performed as a minimum. Testing in triplicate as a minimum provides a level of
statistical confidence in the observed test result.

Testing will be conducted at 0, 3, 6, 9, 12 and 13 months.

Note 2: Testing beyond 13 months will allow an understanding of when, in real-

time, the IVD is likely to ‘fail’ and may allow an extension of the proposed shelf -life.

Note 3: For determination of shelf-life a fresh bottle of specimen buffer must be

opened at each testing point – although there may be circumstances in which
multiple sampling could be taken from the same bottle after it has been opened.
Acquire sufficient volume of each stability testing panel member for the duration
of the testing schedule.

The protocol for these studies specifies the number of devices to be picked, the
statistical sampling plan to be used and the required stability testing panel
members and their volumes.

Documentation:
In Worksheet xxxxx01 record the following:

• The lot numbers from which IVD kits were sampled

• The number of IVD kits sampled from each lot
• Details (including manufacturing/lot information) for each of the IVD kit
components that will be tested as part of this protocol (Test cassette and Sample
Buffer)
The IVD kits chosen to be tested are in their final packaging including all
labelling.
The IVD kits are stored so that the reagents are in contact with all elements of the
packaging (e.g. the bottles in the product kits are stored horizontal lying flat on
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their sides).
The IVD kits will be divided into two groups. One group will be stored at 40 ± 5°C ,
the other
at 8 ± 2°C . Kits from each group will then be subjected to the following conditions.

Testing schedule: for transport simulation

Condition 1, Temperature and humidity sequence: all IVD kits will be taken through
a
temperature and humidity sequence consisting of:
i) Ambient humidity (X% RH)
➢ Put at IFU storage temperature for 24±4 hours followed by
➢ 30 ± 5°C for 24±4 hours followed by
➢ 45 ± 5°C for 24±4 hours, followed by
➢ 8 ± 5°C for 24±4 hours, followed by
➢ IFU storage temperature for 24±4 hours

Followed by

ii) Desert humidity (30% RH)

➢ Put at IFU storage temperature for 24±4 hours followed by
➢ 30 ± 5°C for 24±4 hours, followed by
➢ 45 ± 5°C for 24±4 hours, followed by
➢ 8 ± 5°C for 24±4 hours, followed by
➢ IFU storage temperature for 24±4 hours

Followed by

iii) Tropical humidity (85% RH)

➢ Put at IFU storage temperature for 24±4 hours followed by
➢ 30 ± 5°C for 24±4 hours, followed by
➢ 45 ± 5°C for 72±4 hours, followed by
➢ 8 ± 5°C for 24±4 hours, followed by
➢ IFU storage temperature for 24±4 hours

Followed by

iv) Ambient humidity (X% RH)

➢ Put at IFU storage temperature for 24±4 hours followed by
➢ 30 ± 5°C for 24±4 hours, followed by
➢ 45 ± 5°C for 24±4 hours, followed by
➢ 8 ± 5°C for 24±4 hours, followed by
➢ IFU storage temperature for 24±4 hours

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Note 1: It is important to make clear that the above complete sequence of
temperatures will be used, as opposed to separate IVD kits being held at individual
temperatures. The actual temperatures, durations and the nature of the sequence
will depend on the IVD and the kinds of conditions expected to be encountered
during shipping
Note 2: Freezing temperatures are not considered in this example but should be
included if the IVD kits could be exposed to freezing temperatures during transport.
Note 3: If transport by air is anticipated, the effect of reduced pressure should be
included in the protocol (3) for a period of time at least 10% longer than the longest
anticipated flight, and at a pressure expected in aircraft holds.
Note 4: The protocol should call for testing of at least five individual IVD kits after
each stress condition, using the stability panel members giving the most informative
results. This approach will enable verification that the IVD kits are sufficiently stable
to progress to the next condition, although this should already be known from
preliminary experiments and R&D work.

Condition 2, Transport stress conditions - Shaking. Each IVD kit will be placed on a
shaking table at X revolutions per minute (rpm) for X hours/days at 42 ± 5°C.
After the simulated shipping challenge, each IVD kit will be returned to its
corresponding storage temperature (40±5°C or 8±2°C).

Testing schedule for real time stability studies

Testing will be conducted at 0, 3, 6, 9, 12 and 13 months. At each scheduled time
point, the allotted number of IVD kits will be brought to 15 to 30 °C and used to test
each member of the panel in triplicate.
Note 1: The test at 0 month will provide evidence that the IVD kit is stable under
extreme conditions of shipping (but similar to those likely to be experienced), the
testing at later time points will give evidence to support the claimed shelf life after
transport, and testing beyond the claimed shelf life will provide evidence that the
IVD kit is stable and not close to a failure point.

Documentation for transport stress conditions

In Worksheet XYZ00001 record:
• The lot numbers of the IVD kits used to conduct the test
• The Operator(s) name(s)
• The dates of testing
• Identifying details for each member of the panel being tested
• The temperature at which the IVD kits are stored
• The values of temperature and humidity for each of the challenge conditions
• Instrument settings for the shaking apparatus and duration of operation for
challenge conditions.
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• The ambient temperature and humidity during testing

• Each test result as an interpretation according to the IFU
• Each test result as a band intensity. Band intensity should be scored using
the calibrated scale described in Protocol ZXY0001 (e.g. 0, faint/trace, +1,
+2, +3 … +10) (even though the IFU does not give scores to results)
• Any aberrations or deviations from the protocol, the reason for the deviation
and any remedial action undertaken. Results from invalid assays must be
recorded but not included in calculations of shelf life. Apparently aberrant
results, unless the underlying cause can be positively identified as not
related to a problem with the IVD, must be included in the calculations of
shelf life.

Acceptance criteria
Each panel member should show a band intensity result that matches its expected
result at each tested time point. The expected result must be validated so that if the
IVD fails to meet the claims (e.g. fails to detect critical specimens, has unacceptable
performance at medical decision concentrations, has unacceptable specificity) the
panel member would also fail to meet its specified result.

The stability after transportation of the IVD kit will be taken as the time point before
the last time point to have met the acceptance criteria, e.g. if the IVD is stable to 13
months, the stability after transportation will be deemed to be 12 months.

The stability after transportation should be identical to the claimed shelf life of the
IVD kit, i.e. the extremes of possible conditions to which the IVD kit is likely to
subjected during transport must not affect the shelf life of the IVD.

Calculation of results
Detailed statistical instruction must be obtained from a professional statistician with
an understanding of the expectations of the stability study plan and outcome.
Professional statistical input is particularly recommended when calculating
confidence limits for discrete data such as readings from a graduated scale.

Each of the following applies at each time point:

The variance of the results for all replicates within and between all the lots must be
calculated for each panel member. From the overall variance between lots, the
confidence with which future lots of the IVD kit will detect the panel member at that
time point after manufacture and transport can be calculated. If the confidence of
the panel member meeting its specification is less than some pre-defined value
(normally 95%), it must be deemed to have failed at that time point and the shelf life
of the IVD kit should be restricted accordingly.

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If regression analysis is used to define the time point at which a panel member
would not meet its criterion, then lot-to-lot variability must be included when setting
the confidence limits around the regression line. However, real-time data must
extend beyond the claimed shelf life so that the intercept of the regression
confidence limit and the expected value must be at a time period longer than the
claim. It is usually more appropriate to calculate as discussed in the previous
paragraph, particularly if the regression cannot be proven to be linear.

Example 2: In-use stability protocol

Objective
To determine the stability of opened bottles of the Specimen Buffer used in the IVD
kit in real-time when stored at 15–30°C as proposed in Stability Study Plan
XYZ00001.
In this example, the manufacturer recommends that the test cassette be used
immediately upon opening; this claim should also be validated in a separate
experiment, so that it can be confirmed that the IVD will still perform satisfactorily
after the test cassette has been removed from its pouch and open at room
temperature for 1, 2, 6, 24 hours, etc., as appropriate.

Acquire sufficient numbers of IVD kits from one production lot using a
predetermined sampling protocol (e.g. random, first X number of kits in the first box,
every 100th kit, etc.).

Acquire sufficient volume of each panel member for the duration of the testing
schedule. Establish a method for randomizing the panel for testing.

In Worksheet XYZ00001 record the following:

• The lot numbers from which the IVD kits were sampled
• The number of IVD kits sampled from each lot
• Details (including manufacturing/lot information) for each of the IVD kit
components that will be tested as part of this protocol (test cassette and
specimen buffer).

Preparation

Two lots of specimen buffer are to be tested. One lot of the component must be
freshly made, the other should be towards the end of the assigned shelf life of the
IVD kit.

The component is to be tested in its final packaging.

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The IVD kits are stored so that the reagents are in contact with all elements of the
packaging (e.g. the bottles in the IVD kits are stored horizontally, lying flat on their
sides, allowing liquids to remain in contact with the bottle closures).

Half of each lot will be stored at 30± 5°C, the other half at 15± 5°C. At the start of
testing each bottle will be brought to room temperature (20 ± 2°C), opened, used for
testing and then recapped and returned to the stated storage temperature.

Note 1: It is important that the components under test are opened and used under
circumstances likely to occur in users’ laboratories (i.e. not in rooms with HEPA
filtered air) mimicking, as far as possible, genuine use.

Testing schedule

At each subsequent scheduled time point the allotted number of bottles will be
brought to room temperature and used to test each panel member in triplicate.
Testing will be conducted at 0, 1, 2, 3, 4 weeks, etc., up to the end of the claimed
in-use life.

Documentation

In Worksheet XYZ00001 record:

• The lot number of the IVD kit used to conduct the test
• The Operator(s) name(s)
• The dates of testing
• The temperature at which the IVD kits are stored
• The ambient temperature during testing
• Identifying details for each member of the panel being tested
• Each test result as a band intensity. Band intensity should be scored using
the calibrated scale described in Protocol ZXY0001 (e.g. 0, faint/trace, +1,
+2, +3 … +10)
• Each test result as an interpretation according to the IFU
• Any aberrations or deviations from the protocol, the reason for the deviation
and any remedial action undertaken.
Acceptance Criteria
Each panel member should show a band intensity result that matches its expected
result at each tested time point. The in-use stability of the sample buffer will be
taken as the time point before the last time point to have met the acceptance
criteria.

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Example: If the IVD kit is observed to be stable to 5 weeks, the in-use stability will
be deemed to be 4 weeks.

APPENDIX – II

Suggested specimens for stability testing panels

Examples in this section

Not all of the specimens in the examples that follow will be necessary for all IVDs,
nor is the list exhaustive. Panels must be composed according to strict risk
management principles, and all decisions must be documented and traceable.
The minimum specimens that are recommended to be included in a testing panel
for the different products are outlined below.

1 Specimens to monitor tests for nucleic acid-based testing technology

If a proprietary nucleic acid preparation /extraction system is provided, the recovery

must be shown to meet claims for each genotype from each of the specimen types
claimed (e.g. dried blood spots, whole blood, plasma). Successful removal of
inhibitory substances, if intended, must be demonstrated for appropriate specimen
types. Unless potentially variable biological reagents are involved, this system
would be expected to be verified in manufacture and not necessarily tested at
release.

Specimens Remarks

Specimens to Traceability is required to one of the standard of NIB or

demonstrate NIBSC or WHO or any other, which are required suitably
maintenance of justified on each of the claimed specimen types. More
sensitivity and/or than one genotype may be required to validate these
limit of detection, claims.
and/or accuracy, and
precision

Specimens to Sufficient negative specimens should be included to

demonstrate ensure that the claims will be met at end of shelf life.
specificity and
validity of runs

Specimens (or If more than one part of the genome is to be detected, both
reagents) to systems must be shown to be stable.
demonstrate If both DNA and RNA are measured the complete system
stability of each of must be shown to be stable.
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the critical
components of the
IVD

2 Specimens to monitor tests that measure CD4 cells

Rationale
CD4 measurements are quantitative, and accuracy at the clinical decision points is
important. The design input should have information on the accuracy and other
parameters required, and the panel must be designed to provide evidence that
these parameters are maintained over the assigned life of the reagent and
measuring IVD.

Parameters
The panel used in stability work must be able to demonstrate the following.
• Stability of all the antibodies used in the IVD (frequently anti-CD4 and anti-
CD3 antibodies; any other critical components must also be covered)
• Accuracy and trueness of measurement maintained at the critical level (at
least five specimens required)
• Claimed linearity over the required range of CD4 count (at least five
specimens required)
• Measure drift

Specimens
Artificial specimens, such as stabilized blood specimens, can be used if a risk
assessment based on R&D work indicates that they are effective. Fresh specimens
are usually required. Measurements should be compared to an approved reference
system.

Examples of approaches
Aged or in-use lots may be compared with a reference, e.g. a new lot.

3 Specimens to monitor tests for HIV antibodies

Specimens Remarks

IgM first Possible approaches to obtain samples :

seroconversion • Study the early data from commercial
specimens and IgG seroconversion panels where the seroconversion
first seroconversion was frequently monitored by IgM and IgG blots
specimens • Study the responses to second and third generation
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assays or protein A and protein L assays (this
approach is less useful).

All other parts of the

HIV proteome
included, e.g. reverse
transcriptase (RT)

Late stage specimens This might serve to monitor any kit run control.
– usually a high HIV serology is not particularly genotype dependent. It is
dilution set near the usually not necessary to include controls for genotype
sample-to-cut-off ratio detection unless risk assessment or experiment shows that
it is required for a particular IVD.
HIV-2, diluted to near
the sample-to-cut-off Seroconversion specimens are very rare.
ratio

HIV-1 (0), if claimed

Difficult specimens to Negatives at release subject to risk analysis and statistical

monitor specificity and analysis of the allowable (relative to the claimed) false
invalid rates reactive rate and invalidity rate

4 Specimens to monitor tests for antibodies for HIV-1/2 and Treponema

pallidum (TP)

Specimens Remarks

Specimens to detect See above section 3 Specimens to monitor tests for HIV
HIV antibodies

Note: Each of these epitopes play a role in detecting

Specimens to detect syphilis in different stages of the infection. It is necessary
all the critical epitopes to have a panel member to monitor each epitope system
in the IVD, for present (and possibly each stage of infection), even if poly-
example TpN47, fusion proteins are used. This can be avoided if the
TpN17 and TpN15 manufacturer can demonstrate that each epitope system is
equally stable.

Specimens able to Note: It would not be sufficient for CDSCO to extrapolate to

show that the the stability of HIV-2/TP detection by testing only HIV-1
invalidity and positive specimens.
specificity rates do not
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fall outside the claims,
particularly if whole
blood is a claimed
specimen type

5 Specimens to monitor tests for Hepatitis C virus antibodies

Specimens Remarks

NS3 first
seroconversion
specimens and core
first seroconversion
specimens

Specimens to monitor
any other antibodies Results can be obtained from line immunoassays that
claimed (frequently differentiate antibody responses to the different proteins.
against NS5 and NS4)

Note: Hepatitis C virus serology is not particularly genotype

dependent in terms of anti-core and anti-NS3, but it is
A late-stage dilution possible to make serotyping assays based on NS4 that
near the sample-to- mimic genotyping reasonably well. It is usually not
cut-off ratio necessary to include controls for genotype detection,
unless risk assessment or experiment for a particular IVD
show otherwise.

Difficult specimens to Negative specimens subject to risk analysis and statistical

monitor specificity and analysis of the allowable false reactive rate and invalidity
invalid rates rate (relative to the claimed rates)

6 Specimens to monitor for tests for Hepatitis B surface antigen (HBsAg)

Specimens Remarks

Specimens to define Traceability is required to one of the standard of NIB or

sensitivity relative to NIBSC or WHO or any other, which are required suitably
the claim justified on each of the claimed specimen types.

Specimens to monitor These will almost certainly be traceable to the “First WHO
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the maintenance of International Reference Panel 2011, for Hepatitis B virus
the claims of a variety genotype panel for HBsAg-based assays" PEI code:
of serotypes / 6100/09.
genotypes and mutant
forms

Specimens to control
against prozone/high
dose hook effect if
found or if
theoretically an issue

If detection of HBsAg
in the presence of
anti-HBsAg is claimed
(current best practice)
proof of maintenance
of the claim

If the monoclonal antibodies used have particular function

or bias, such as against the ayr or adr subtypes (not
Specimens to monitor controlled by the standards) or to detect mutant forms of
the critical the antigen, each must be monitored to ensure viability at
components of the end of shelf life. These may be the same specimens as
IVD mentioned in the previous paragraphs.
If there are critical dissociation chemicals or red-cell
capture or rupture agents used, these must also be
monitored.
Difficult specimens to Negatives subject to risk analysis and statistical analysis of
monitor specificity and the allowable (relative to the claimed) false reactive rate
invalid rates and invalidity rate.

CDSCO/IVD/GD/Stability/02/2022 Page 47 of 50
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
APPENDIX – III

Examples of Stability Study approaches

1 Accelerated Stability Study: (Three lots)

Product Claimed Claimed Accelerated Testing

name Storage shelf life condition interval
condition
HIV 1-2 Ab 2-30°C 24 months 2-8°C, Room 0 day, 4thday,
Rapid Temperature (20- 7thday,
qualitative test 30°C), 37°C and 14thday,
Rapid 45°C 21stday,
qualitative test 30thday and
41stday

2 Real Time Stability Study: (Three lots)

Product name Claimed Claimed Real Time Testing

Storage shelf life Stability interval
condition condition
HIV 1-2 Ab Rapid 2-30°C 24 months 2-8°C, Room 0 month (Initial
qualitative test Temperature Testing) and
Rapid qualitative (20-30°C), 3 months, up
test to 27 months in
three months
interval

CDSCO/IVD/GD/Stability/02/2022 Page 48 of 50
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India
3 In Use Stability Study: One lot

Product Claimed Claimed Claimed Claimed In Use Testing

name Storage shelf In Use Stability Stability or interval
condition life Stability after Open
or Open opening Bottle
Bottle Stability
Stability condition
condition
HIV 1-2 2-30°C 24 2-30°C up to 24 2-8°C, Room 0 hrs
Ab Rapid months hours Temperature (initial
qualitative after (20-30°C), testing),
test Rapid opening 37°C and then at
qualitative 45°C 0 hrs
test (initial
testing),
then at 0
hr, 1 hr,
2 hrs, 3
hrs , 4
hrs,
5hrs, 6
hrs,
24hrs
and at
25hrs

CDSCO/IVD/GD/Stability/02/2022 Page 49 of 50
 Central Drugs Standard Control Organization
Directorate General of Health Services,
Ministry of Health and Family Welfare, Government of India

4 Shipping stability study: One lot

Product name Claimed Claimed Control Test group

Storage shelf life group temperature and
condition without Humidity cycle
temperature
and Humidity
cycle
HIV 1-2 Ab 2-30°C 24 2-30°C Cycle:1 at 40% RH
Rapid months 1-40°C for 24hrs ± 4
qualitative test Followed 30 ± 5 °C
Rapid for 24hrs ± 4
qualitative test Followed 45 ± 5 °C
for 24hrs ± 4
Followed1-40°C for
24hrs ± 4
Followed
Cycle:2 at 85% RH
1-40°C for 24hrs ± 4
Followed 30 ± 5 °C
for 24hrs ± 4
Followed 45 ± 5 °C
for 72hrs ± 4
Followed1-40°C for
24hrs ± 4
Followed
Cycle:2 at 40% RH
Followed 30 ± 5 °C
for 24hrs ± 4
Followed 45 ± 5 °C
for 24hrs ± 4
Followed1-40°C for
24hrs ± 4

CDSCO/IVD/GD/Stability/02/2022 Page 50 of 50