OUT OF SPECIFICATION (OOS) RESULTS
Kiran Kota
OOS is the test result that does not comply with the pre-determined acceptance criteria (i.e. for example, filed applications, drug master files, approved marketing submissions, or official compendia or internal acceptance criteria).
Regulatory Reference Guidelines:
Stability studies on marketed batches of finished products and or active pharmaceutical ingredients, on-going / follow up
Batch release testing & testing of starting materials & batches for clinical trials
Phase-I (Lab Investigation)
Responsibility of an
Analyst
Responsibility of Lab
Supervisor
OUT OF SPECIFICATION (OOS) RESULTS
Investigating Out-of-Specification (OOS) Test Results for
Investigating Out-of-Specification (OOS) Test Results
Pharmaceutical Production Guidance for Industry
Previous released batch used as reference sample in an OOS investigation showing OOS or suspect results
stability (no stress tests)
OOS Investigation
shall be done if Data is used for batch calculations/decisions and if in a dossier and on Certificates of Analysis
IDENTIFICATION AND ASSESSMENT OF OOS RESULTS
Phase-I
(Lab Investigation)
KIRAN KOTA
Phases of OOS investigation
Phase-II
(Full scale Investigation)
Phase-Ia
Phase-I
(Laboratory Investigation)
(To identify Obvious
error)
Phase-Ib
(To identify assignable
cause)
Phases of OOS
investigation
Phase-II
(Manufacturing Investigation)
Phase-III
(When batch is rejected / No cause
identified from Phase-I & II)
Phase-I Lab Investigation
Phase-I includes initial assessment of accuracy of laboratory data
This shall be done before test preparations are discarded, determines laboratory error
Phase-Ib
Phase-Ia
or instrument error.
To determine whether there
If no out come full scale investigation shall be carried out.
Contract lab should notify all the data to manufacturing firms quality unit, which has been a clear obvious errors
should initiate full scale investigation. ( i.e. Calculation error , Power Initial Investigation conducted by
outage , Equipment failure , the analyst and supervisor using
Testing errors, Incorrect the Laboratory Investigation
Instrument Parameters) Checklist (which containes check
• Should aware on potential problems that could occur. For microbiological analysis ponts like Correct method, correct
• Ensure appropriate & Calibrated instrumets are used. this may be after the analysis sample, calibrated instruments,
• Analytical methods not meeting the System Suitability should not be used. has been completed and Correct standards, clean glassware
• Data should be checked for compliance before discarding of test preparations. reviewed during reading of the used for analysis and done by
• If unexpected results with no obvious error, the test preparations sholub be samples. Qualified analyst, checked the SST
retained if stable.
parameters)
• If abvious error, analyst should immediately document what happened.
Contact Production/Contract
If Obvious error identified Giver/QP/MAH as appropriate
(with justification), invalid the
initial analysis and batch On completion of the Analyst and
analysis shall be performed Supervisor investigation re-
• Data should be assessed promptly to identify lab error or manufacturing error. measurement can start once the
• Discuss with analyst, examine raw data, verifying calculations, instrument
calibrations, verifying standards, past data of test performance. hypothesis plan is documented and
• Assessment of cause is facilitated if retained sample preparations are examined If no error was noted, and none is only to support the
promptly. of the above conditions were investigation testing.
• hypothesis should be tested. met Phase Ib investigation must
• Examination of retained solutions. take place.
• With clear evidence of lab error, initial data to be invalidated. If evidence is
unclear, full scale investigation to be carried out.
 OUT OF SPECIFICATION (OOS) RESULTS
Kiran Kota
OUT OF SPECIFICATION (OOS) RESULTS
Phase-II Investigation Phase-II Investigation

8Phase-II investigation conducted when
the phase-I investigations did not reveal an
8Should be conducted by cross functional team. assignable laboratory error.
8For contract testing: The contract testing lab personnel should be involved. 8Phase-II investigations are driven by
Review of 8Timely, thorough & well documented review should include clear statement for reason of investigation, written and approved instructions against
production Summary of manufcaturing process aspects which may have caused the problem. hypothesis.
8If investigation confirms OOS results, investigation should be terminated and batch rejected. 8Prior to further testing a manufacturing
8Investigation should extended to other batches / product impacted investigation should be started to
determine whether there was a possible
manufacturing root cause.
Hypothesis Testing (Applicable to Phase-Ia and Phase-II)
8Should be started as part of Phase Ia and continue into Phase II if no assignable cause found.
8Description of the testing should be written, and then approved by QA/Contract Giver/QA equivalent
prior to initiating investigational testing.
8This Hypothesis testing may continue from the re-measurement of the original preparations.
8 Investigational testing may not be used to replace an original suspect analytical results.
8It may only be used to confirm or discount a probable cause
8If no assignable cause that could explain the results can be identified during the manufacturing
investigation or the assay failure investigation retesting may be considered.
8Part of the investigation may involve retesting a portion of the original sample.

KIRAN KOTA
Averaging
8The validity of averaging depends upon the sample and
its purpose.
8Using averages can provide more accurate results
8Averaging cannot be used in cases when testing is
intended to measure variability within the product, such as
powder blend/mixture uniformity or dosage form content
Re-testing: uniformity
8Sample should be taken from same homogenous lot which resulted in OOS. 8Reliance on averaging has the disadvantage of hiding
variability among individual test results. For this reason,
8Determine instrumet/sample handling error.
all individual test results should normally be reported as
8Retesting should be based on scientific rationale.
separate values.
8Number if retestings, should be mentioned in SOP. 8Where averaging of separate tests is appropriately
8If additional testing required, to be done on protocol basis. specified by the test method, a single averaged result can
8If clear lab error, retest data substituted with original results. be reported as the final test result.
Additional 8If no lab error both initial and retest results shall be documented for batch disposition decission. 8In some cases, a statistical treatment of the variability
Laboratory of results is reported
Retesting
8Performed on the original sample not a different sample.
8Can be a 2nd aliquot from the same sample that was the source of the original failure.
8If insufficient quantity of the original sample remains to perform all further testing
then the procedure for obtaining a resample must be discussed and agreed by
QA/Contract Giver/QA equivalent.
8The process of obtaining the resample should be recorded within the laboratory
investigation.
8The decision to retest should be based on sound scientific judgement. The test plan
must be approved before re testing occurs.
8The minimum number of retests should be documented within the procedure and be
based upon scientifically sound principles.
8The number of retests should be statistically valid; suggested 5, 7, or 9.
8The retests should be performed by a different analyst where possible.
8The second analyst should be at least as experienced and qualified in the method as
the original analyst.

testing
Re-sampling
Re-sampling:
8Should rarely occur
8Analysing specimen sample collected by same method or additional sample collected from Retesting should
8If insufficient quantity of the original sample remains to perform all further testing
be done by another analyst. then the procedure for obtaining a resample must be discussed and agreed by
8Such sampling & testing can conclude that original sample was not properly prepared & hence not QA/Contract Giver/QA equivalent.
represent quality of the batch. 8The process of obtaining the resample should be recorded within the laboratory
8It determines that original sampling method was inadequate, new accurate method should be developed. investigation.
8Re-sampling should be performed by the same qualified methods that were used for
the initial sample. However, if the investigation determines that the initial sampling
method was in error, a new accurate sampling method shall be developed, qualified and
documented.
8It involves the collecting a new sample from the batch.
8Will occur when the original sample was not truly representative of the batch or there
was a documented/traceable lab error in its preparation.
8Evidence indicates that the sample is compromised or invalid. Sound scientific
justification must be employed if re-sampling is to occur
Averaging:
8Appropriate Use: Use of replicates to arrive at single result & number of replicates used should be specified.
Acceptance limit for variability among replicates should be defined.
Averaging should be used only if it is used during original analysis. Phase-III Investigation
Outlier test:
8An outlier may result from a deviation from prescribed
test methods, or it may be the result of variability in the
sample.
8It should never be assumed that the reason for an
outlier is error in the testing procedure, rather than
inherent variability in the sample being tested.
8Statistical analysis for Outlier test results can be as part
of the investigation and analysis.
8However for validated chemical tests with relatively
small variance and that the sample was considered
homogeneous it cannot be used to justify the rejection of
data.
8While OOS guidance is not directly intended for bioassay
analysis, it can be used as a starting point for the
investigation.
8Compendia such as the BP; PhEur and USP, provide
guidance on outliers for these types of analysis

8Inappropriate use: Averaging has potential of hiding variability in test results. All individual test should be
reported as individual result. Use of averaging is inappropriate during powder blend/mixture uniformity or
8The phase 3 investigation should review the completed 8Once a batch has been rejected
dosage form content unifirmity as test is intended to measure variability within the product there is no limit to further testing
If additional testing is done during OOS investigation, averaging should not be done with original result. Relying manufacturing investigation and combined laboratory
investigation into the suspect analytical results, and/or to determine the cause of
on average is misleading when some results are OOS and other are within specifications. 8If the batch is rejected there failure, so that corrective action
Reporting test method validation for possible causes into the results
still needs to be an can be taken.
results obtained.
investigation. 8The decision to reject cannot
8To conclude the investigation all of the results must be
be reversed as a result of further
evaluated.
To determine: testing.
Outlier tests: 8The investigation report should contain a summary of
8If other batches or products 8The impact of OOS result on
8There should be a statistically valid QC criteria with appropriate acceptance or rejection level. the investigations performed; and a detailed conclusion.
are affected. other batches, on going stability
8In some cases , a value may be obtained which is different from others in a seies due to deviation of test 8For microbiological investigations ,where appropriate,
studies, validated processes and
method or variability in sample. use risk analysis tools to support the decisions taken and
8identification and testing procedures should be
8There should be no assumption related to error in test method. conclusions drawn. It may not have been possible to determined by Quality Control
implementation of corrective
8Outlier testing identifies extreme from a data, this testing should be done in advance as per a defined SOP. determine the actual root cause therefore a robust most and Quality Assurance and be
and preventative action.
8Minimum number of results required to obtain statiscally significant assessment for outlier test should be probable root cause may have to be given. documented in the conclusion,
defined in the SOP. 8The batch quality must be determined and disposition along with appropriate corrective
8QC or contract testing testing lab shall provide all the individual test results to QA for batch disposition decision taken. and preventive actions

Results should be evaluated and batch quality should be determined, release decision should be made by QA.
Once batch is rejected , there is no limit for future testing to determine root cause for determine CAPA.
Intrepreting results
8Quality unit is responsible to
interpretation of results.
8An OOS doesnot mean the batch is
failure and it should be rejected.
8Investigation must be carried to
interpreted to determine the result
and batch disposition.
8When an investigation reveals a
cause, the OOS result is invalidated
and not used to determine the
quality of the batch, Investigation
should be done on confirmation of an
event that has caused an OOS.
8When investigation reveals that
OOS is caused by factors affecting
batch quality, result should be used
in evaluting the batch quality and
batch should be rejected.
8The investigation should be
extended into other batches and
products associated.
8When investigation does not
reveals cause and does not confirm
OOS, results should be given full
consideration in disposition decision.
8In this case Quality Unit can
release the batch and should include
appropriate follow-up and security
to prevent lab error that could have
led to OOS.
8The final descision to release the
batch should comes only when full
investigation concludes that the
OOS results does not represent the
quality of the batch.
OUT OF SPECIFICATION (OOS) RESULTS
Cautions
KIRAN KOTA
Averaging results from multiple sample
preparation from original sample:
8For Assay results, if some individual
test are with in specification and some
OOS and both within variability, then
within specification results doesnot
represent true quality of the sample.
8It should be cautious and consider the
average as OOS, even if the average is
within specification.
8As per USP general chapter, every
individual application of official test
should be within specification.
Averaging results from same final
sample preparation :
8There may be causes where test
method specifies appropriate
acceptance criteria for vaiability and pre-
defined number of replicates from final
diluted sample solution to arrive the test
result.
Borderline results that are within
specification:
8An assay result that is low, but within
specifications, should also raise a
concern.
8One cause of the result could be that
the batch was not formulated properly.
8Batches must be formulated with the
intent to provide not less than 100
percent of the labeled or established
amount of active ingredient.
8As with all analytical testing
conducted to evaluate the quality of a
drug, all records pertaining to the OOS
test result should be retained.
OUT OF SPECIFICATION (OOS) RESULTS
CONCLUDING THE INVESTIGATION
Field Alert Reports
8For those products that are the
subject of an approved new drug
application or abbreviated new
drug application, regulations
require submitting within
working days a field alert report
(FAR) of information concerning
any failure of a distributed batch
to meet any of the specifications
established in an application
8OOS test results on these
products are considered to be one
kind of "information concerning
any failure” described in this
regulation. Unless the OOS result
on the distributed batch is found
to be invalid within 3 days, an
initial FAR should be submitted. A
follow-up FAR should be submitted
when the OOS investigation is
completed.
Kiran Kota
If Obvious error identified (with justification), invalid the initial analysis and batch analysis shall be
performed.
If no laboratory or calculation errors are identified in the Phase I and Phase II there is no scientific
basis for invalidating initial OOS results in favour of passing retest results. All test results, both
passing and suspect, should be reported (in all QC documents and any Certificates of Analysis) and all
data has to be considered in batch release decisions.
If the investigation determines that the initial sampling method was inherently
inadequate, a new accurate sampling method must be developed, documented, and
reviewed and approved by the Quality Assurance responsible for release. A consideration
should be given to other lots sampled by the same method.
An initial OOS result does not necessarily mean the subject batch fails and must
be rejected. The OOS result should be investigated, and the findings of the
investigation, including retest results, should be interpreted to evaluate the batch
and reach a decision regarding release or rejection which should be fully
documented.
3
In those cases where the investigation indicates an OOS result is caused by a
factor affecting the batch quality (i.e., an OOS result is confirmed), the result
should be used in evaluating the quality of the batch or lot. A confirmed OOS
result indicates that the batch does not meet established standards or
specifications and should result in the batch's rejection and proper disposition.
Other lots should be reviewed to assess impact.
For inconclusive investigations — in cases where an investigation:-
(1) does not reveal a cause for the OOS test result and
(2) does not confirm the OOS result
The OOS result should be given full consideration (most probable cause determined)
in the batch or lot disposition decision by the certifying QP and the potential for a
batch specific variation also needs considering.
Any decision to release a batch, in spite of an initial OOS result that has not been invalidated, should
come only after a full investigation has shown that the OOS result does not reflect the quality of the
batch. In making such a decision, Quality Assurance/QP should always err on the side of caution.