PHARMA DEVILS

QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

1.0 PURPOSE
To define the characteristics for consideration during the validation of an analytical method of drug
product (Pharmacopoeial or in-house).

2.0 SCOPE
2.1 This procedure describes the typical characteristics to be selected for validation of an analytical method,
the process for their determination and the acceptance criteria applicable.

3.0 REFERENCE(S) & ATTACHMENTS

3.1 References
3.1.1 Validation of Analytical Procedures: Text and Methodology – Q2 (R1)
3.1.2 Chapter <1225> Validation of Compendial Procedures; USP.
3.1.3 Chapter <1092> The Dissolution procedure: Development and Validation.

3.2 Attachments
3.2.1 Nil

4.0 DEFINITION & ABBREVIATION(S)

4.1 Definitions
4.1.1 Specificity:
Specificity is the ability of the analytical method to assess unequivocally the analyte in the presence of
components that may be expected to be present in the sample.
4.1.2 Limit of detection:
The detection limit of an individual analytical procedure is the lowest amount of analyte in a sample,
which can be detected but not necessarily quantitated as an exact value (s).
4.1.3 Limit of quantitation:
The Quantitation limit of an individual analytical procedure is the lowest amount of analyte in a sample,
which can be quantitatively determined with suitable precision and accuracy.
4.1.4 Linearity:
The linearity of an analytical procedure is its ability (within a given range) to obtain test results that are
directly proportional to the concentration of analyte in the sample.
4.1.5 Precision:
The precision of an analytical procedure expresses the closeness of agreement (degree of scatter)
between a series of measurements obtained from multiple samplings of the same homogeneous sample

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QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

under the prescribed conditions.

4.1.6 Repeatability:
Repeatability expresses the precision under the same operating conditions over a short interval of time.
Repeatability is also termed intra-assay precision.
4.1.7 Intermediate Precision:
Intermediate precision expresses within-laboratories variations: different days, different analysts,
different equipment, etc.
4.1.8 Accuracy:
The accuracy of an analytical procedure expresses the closeness of agreement between the value,
which is accepted either as a conventional true value or an accepted reference value and the found value.
4.1.9 Robustness:
The robustness of an analytical procedure is a measure of its capacity to remain unaffected by small, but
deliberate variations in method parameters and provides an indication of its reliability during normal
usage.
4.2 Abbreviations
4.2.1 GC: Gas Chromatography
4.2.2 HPLC: High Performance Liquid Chromatography
4.2.3 LOQ: Limit of Quantification
4.2.4 LOD: Limit of Detection
4.2.5 QA: Quality Assurance
4.2.6 QC: Quality Control
4.2.7 R&D: Research and Development
4.2.8 RSD: Relative Standard Deviation
4.2.9 Spec.: Specifications
4.2.10 STP: Standard Testing Procedure
4.2.11 UV: Ultra Violet.

5.0 RESPONSIBILITY
5.1 Corporate Quality Assurance
5.1.1 To ensure implementation of the procedure
5.2 Quality Control Analyst:
5.2.1 To ensure availability of required reagents, working standard/ reference standard/ column/ instrument.
5.2.2 To perform and record the analytical method validation.

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 PHARMA DEVILS
QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

5.3 Head QC or designee

5.3.1 To review and ensure correct analytical method validation.
5.3.2 To provide the required reagents/material/column/instruments.
5.3.3 To approve the analytical method validation report.
5.3.4 To ensure implementation of system as per guideline.
5.4 Quality Assurance Head:
5.4.1 To ensure implementation of procedure.

6.0 Distribution:
I. Quality Assurance
II. Quality Control

7.0 PROCEDURE:
7.1 ANALYTICAL METHOD VALIDATION FOR ASSAY/PRESERVATIVE TEST:
7.1.1 Parameters to be considered for an In-house method
7.1.1.1 Specificity
7.1.1.2 Solution stability
7.1.1.3 Filter compatibility
7.1.1.4 Filter saturation
7.1.1.5 Linearity and range
7.1.1.6 Precision (System precision, Repeatability, Intermediate precision)
7.1.1.7 Accuracy
7.1.1.8 Robustness
7.1.1.9 System suitability

7.1.2 Parameters to be considered for a Pharmacopoeial method

7.1.2.1 Specificity
7.1.2.2 Solution stability
7.1.2.3 Filter compatibility
7.1.2.4 Filter saturation
7.1.2.5 Precision (System precision, Repeatability, Intermediate precision)
7.1.2.6 Linearity and range
7.1.3 SPECIFICITY:

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QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

7.1.3.1 Interference study:

Determination:
Prepare and analyse blank solution (diluent), placebo solution, known impurities solutions (as included
in specifications) at specification limit, process impurities according to the peak response, sample
solution and standard solution to check the interference. For preservative test perform only placebo
interference.
For multiple strength, select
• Higher strength if dosage formula is linear
• If in case, different colours of material used for coating and/or formulation, inject each placebo
or placebo mixed with all colours and measure the absorbance /peak responses.
Acceptance criteria:
For HPLC method
No peak shall be observed due to blank solution, placebo solution and known impurity solution at the
retention time of principle peak as observed in the standard solution and sample solution.
For UV method
Interference due to placebo solution at the wavelength of determination shall not be more than 2.0%.
7.1.3.2 Forced degradation study (For HPLC methods):
Perform forced degradation studies to demonstrate that the analytical method is stability indicating.
Follow the respective guideline for forced degradation study for further details.
For multiple strength, select
• Higher strength sample shall be use for degradation if it is linear dosage form
• Lower Drug placebo ratio of dosage form if average weight is same for all strengths.
• If different colours of material used for coating and/or formulation, subject to degradation study
of individual placebo and sample shall be of higher strength.
Acceptance criteria:
The peak of interest shall be spectrally pure and no co-elution shall be observed with blank peak(s) and
/or impurity peak(s).

7.1.4 SOLUTION STABILITY:

Determination:
Prepare standard and sample solutions as described in the method to be validated.
If multiple strength, select

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QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
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Issue Date: Page No.:

• Higher strength if dosage formula is linear

• Lower Drug placebo ratio of dosage form if average weight is same for all strength.

Keep the prepared solutions tightly closed and store at room temperature and at 10°C. Analyze the
standard and sample solution up to 24 h of both the condition. (If possible 48 h), if the drug product
tends to be stable in solution (Based on the information obtained from development report).
Analysis may also be done at intermediate time intervals. However, report the results of initial and
actual time point.
Determine the absolute percentage difference in assay/response at a particular time point with respect
to the initial assay/response forstandard solution and test solution.
Acceptance criteria (For HPLC & UV methods):
The absolute % difference in assay/ response with respect to initial at each time point shall not be more
than 2.0.
If it is outside the set criteria, make appropriate recommendations.

7.1.5 FILTER COMPATIBILITY:

Determination:
Prepare the sample solution as described in the method to be validated.
If multiple strength, select higher strength if dosage form is linear.
At the filtration stage, filter the sample solution through Whatman GF/C filter, Whatman No.41,
Whatman No.42; discard about 10 mL sample solution from each filter.
For 0.45m membrane filter and 0.45µm PVDF discard about 5 mL sample solution, collect the
sample solutions for further analysis.
Centrifuge the same (unfiltered) sample solution. Analyse all the solutions as described in the test
procedure and calculate the assay results.
Determine the absolute difference in the results obtained for the filtered solutions and the centrifuged
solution.
Acceptance criteria (For HPLC & UV method):
The absolute difference in assay results obtained for the filtered solutions and the centrifuged solution
shall not be more than 2.0. If it is out of the set criteria, make appropriate recommendations.

7.1.6 FILTER SATURATION:

Determination:

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 PHARMA DEVILS
QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

Prepare the sample solution as described in the method to be validated.

If multiple strength, select higher strength if dosage form is linear.
At filtration stage, when Whatman filter paper no.41, Whatman filter paper no.42 and GF/C filter is
recommended for use in Filter compatibility study filter 10.0 mL and 20.0 mL sample solution and
discard using three separate filters, followed by filtration of further 10 mL aliquots. Collect the
filtrates each in separate test tubes. When 0.45µm PVDF filter and 0.45µm nylon membrane filter
is recommended for use in Filter compatibility study filter suitable volume (1mL, 3mL and 5mL) of
sample solution and discard using separate filters, followed by filtration of further 10 mL aliquots.
Collect the filtrates each in separate test tubes.
Analyse the thus obtained solutions as described in the method to be validated and calculate the
assay results. Determine the absolute difference in the assay result obtained in two consecutive
aliquots.

Acceptance criteria (For HPLC & UV methods):

The absolute difference in the assay result obtained for two consecutive aliquots shall be not more than
2.0. If it is out of the set criteria, make appropriate recommendations.
7.1.7 LINEARITY AND RANGE:
Determination:
Prepare linearity solutions from the stock solution of standard, to obtain the solutions at 50 % to 150 %
of the working concentration by preparing minimum five concentrations. Inject all the prepared
solutions in duplicate.
Plot a graph of corrected peak areas vs. concentration (ppm). Determine and report the slope, intercept,
and correlation coefficient of the regression line and residual sum of squares.
For range, record the concentration levels over which the results are linear.
Acceptance criteria (For HPLC & UV methods):
Correlation Coefficient shall be not less than 0.999

7.1.8 PRECISION:
7.1.8.1 System precision:
Determination:
Prepare the standard solution as described in the method to be validated and inject the obtained
solution in six replicates. Calculate the relative standard deviation of the responses.
Acceptance criteria (For HPLC & UV methods):

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 PHARMA DEVILS
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STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
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Issue Date: Page No.:

Relative standard deviation shall not be more than 2.0 %

7.1.8.2 Repeatability:
Determination:
Repeatability shall be performed in the following way, for multiple strengths.
• If Sample used for crushed powder for sample preparation, select only higher
strength.
• If sample used for intact tablets for sample preparation, all strength to be performed.
• If dilution of different strength varies, all the strength to be performed.
Prepare six different sample solutions, as directed in the method to be validated, from the same
homogeneous sample and analyse over a short period of time by the same analyst, on the same
equipment and on the same day.
Calculate the assay results. Determine the relative standard deviation and 95 %
Confidence interval of the assay results obtained from the six preparations.
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation of the assay results obtained from six preparations shall not be more than
2.0 %.
7.1.8.3 Intermediate precision:
Determination:
Prepare six different sample solutions, as directed in the method to be validated, from the same
homogeneous sample and analyzed by a different analyst, on different equipment, on different day.
Calculate the assay results. If multiple strength, select only higher strength. Determine the relative
standard deviation and 95 % Confidence interval of the assay results obtained from the six
preparations.
Calculate the absolute difference in the assay results obtained in Repeatability (mean value of six
results) and Intermediate precision (mean value of six results).
Acceptance criteria (For HPLC & UV methods):
• Relative standard deviation of the assay results obtained from six preparations shall be not more
than 2.0 %
• The absolute difference in the assay results obtained in the Repeatability study (mean value of
six results) and Intermediate precision study (mean value of six results) shall be not more than 2.0.
7.1.9 ACCURACY (RECOVERY):
Determination:

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Department: Quality Assurance SOP No.:
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Issue Date: Page No.:

Prepare recovery solutions by spiking the drug substance in to the volumetric flask containing placebo
powder, to obtain the solutions at 50 %, 100 % and 150 % of target concentration of drug substance as
in sample solution described in the method to be validated.
If multiple strength with different colour, select worst condition of placebo as below and perform the
accuracy.
• Lower Drug: placebo ratio if average weight of the dosage basis is same.
• If different colour of material is used for formulation and /or coating material’s mixed with excipient
proportionally in the individual dosage form.
The quantity of placebo in recovery solutions at 50 %, 100 % and 150 % shall remain constant as in
sample solution described in the method to be validated. If the amount of drug substance to be spiked
is less than 10 mg, then prepare a stock solution of the drug substance and use this for spiking to
prepare the recovery solutions. Prepare the recovery solutions at all the three concentration levels in
triplicate with duplicate injection and analyse as per the method to be validated.
Calculate the quantity recovered in mg or µg and the % recovery, for each level and the mean recovery
for all nine solutions.
Acceptance criteria (For HPLC & UV methods):
All the individual recoveries shall be within 97.0 % to 103.0 % and the mean recovery shall be within
98.0 % to 102.0 %.

7.1.10 ROBUSTNESS:
Determination:
Carry out one set of analysis, using the same homogeneous sample. If multiple strength, select only
higher dosage form. By making individual small deliberate changes in the analytical procedure. Select
the changes to be made in the analytical procedure from the below list, as applicable.
• Change in pH of buffer (pH specified in method  0.2).
• Change in pH of mobile phase (pH specified in method  0.2).
• Change in mobile phase composition of each component (Absolute 2 % or 30% relative
whichever is larger).
• Change in column oven temperature (Temperature specified in method  5°C).
Calculate the result of assay for each set of analysis. Determine the absolute difference in the results
obtained in Robustness study and Repeatability study (sample 1).

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 PHARMA DEVILS
QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

Acceptance criteria (For HPLC & UV methods):

• The absolute difference in the results obtained in robustness study and repeatability study
(sample 1) shall not be more than 2.0.
• Shall meet all the requirements of system suitability.

7.1.11 SYSTEM SUITABILITY

Determination:
Perform System suitability before performing any parameter.
Acceptance criteria (For HPLC & UV methods):
The system suitability shall comply as per methodology.

7.2 ANALYTICAL METHOD VALIDATION FOR RELATED SUBSTANCES TEST OF DRUG

PRODUCT:
7.2.1 Parameters to be considered for In-house method
7.2.1.1 Specificity
7.2.1.2 Limit of detection and Limit of Quantitation
7.2.1.3 Solution stability
7.2.1.4 Filter compatibility
7.2.1.5 Filter saturation
7.2.1.6 Linearity and range
7.2.1.7 Precision (System precision, Repeatability, Intermediate precision)
7.2.1.8 Accuracy
7.2.1.9 Robustness
7.2.1.10 Relative response factor
7.2.1.11 System suitability

7.2.2 Parameters to be considered for Pharmacopoeial method

7.2.2.1 Specificity
7.2.2.2 Solution stability
7.2.2.3 Filter compatibility
7.2.2.4 Filter saturation
7.2.2.5 Precision (System precision, Repeatability, Intermediate precision)
7.2.2.6 Limit of Quantitation

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 PHARMA DEVILS
QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

7.2.3 SPECIFICITY:
7.2.3.1 Interference Study:
Determination:
Prepare blank solution (diluent), placebo solution, known impurities solutions at specification limit,
process impurities according to the peak response standard solution and sample solution to check for
interference.
In case of multiple strength dosage form inject all placebo and sample preparation individually and
check the interference study.
Acceptance criteria:
The peaks due to blank solution, placebo solution, principal peak and process impurities shall not
interfere at the retention time of any of the known impurities (those included in the specification),
unknown impurities and main peak and all the known impurity peaks (those included in the
specification) shall be well resolved from each other.
7.2.3.2 Forced degradation study (Applicable to HPLC method):
Perform forced degradation studies to demonstrate that the analytical method is stability indicating.
Follow the respective guideline for the forced degradation study for further details.
For multiple strength, select
• Higher strength sample shall be used for degradation if linear dosage form
• Lower Drug placebo ratio of dosage form if average weight is same for all strength.
• If different colours are used in dosage form, all individual and/or formulation, subject to
degradation study of individual placebo and sample shall be of higher strength.
Acceptance criteria:
The peaks of interest shall be spectrally pure and no co-elution shall be observed with blank peak(s)
and/or impurity peak(s).

7.2.4 Limit of Detection and Limit of Quantitation Determination:

Either of the following shall determine LOD & LOQ values.
7.2.4.1 Signal to noise ratio Method:
Signal to noise is performed by comparing measured signals from sample with known low
concentration of analyte with that blank sample and establishing minimum concentration at which the
analyte can be reliably detected or quantified.
Acceptance criteria:

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Department: Quality Assurance SOP No.:
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Signal to noise ratio is 10:1 for LOQ and 3:1 for LOD.
7.2.4.2 Standard deviation of response and slope method:
Prepare a series of solutions by quantitative dilutions of the stock solution of standards to obtain solutions of
suitable concentrations (at least six different concentrations) from 5 % to 30 % of specification limit. However,
when poor peak response is observed, solutions may be prepared up to 50 % of specification limit. Inject each
solution into the chromatograph in duplicate (inject only once if impurity solutions are not stable) and calculate
the mean peak areas and corrected peak areas.
Determine the slope and residual standard deviation for each standard using the corrected peak areas and
concentration (ppm). Calculate the value of limit of detection and limit of quantitation for each peak using the
following formula:

Calculation:
3.3 x  10 x 
LOD = --------- LOQ = ---------
S S

Where,
 = Residual Standard Deviation
S = Slope
LOD = Limit of detection
LOQ = Limit of quantitation
Prepare a solution at LOQ level for each standard and inject in six replicates. Calculate the relative standard
deviation of the peak areas for each peak.
Acceptance criteria:
Relative standard deviation of the peak areas due to each LOQ level standard injected in six replicates shall not be
more than 10.0 %.

7.2.5 SOLUTION STABILITY:

Determination:
• If no known impurity is given in the specification, prepare sample solution as it is.
• If known impurity is given in the specification and is above LOQ level, prepare sample solution
as it is.

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Department: Quality Assurance SOP No.:
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Issue Date: Page No.:

• If known impurity is given in specification and is below LOQ level, spike the impurity at the
specification limit in the sample solution.
Prepare the standard solutions and sample solution as described in the method to be validated. If multiple strength,
select
• Higher strength sample shall be used if linear dosage form
• Lower Drug placebo ratio of dosage form if average weight is same for all
strength.
Keep the prepared solutions tightly closed and store at room temperature and at 10°C. Analyse the
standard and sample solution up to 24 hours of both the condition (if possible 48 h).
If the drug product tend to be stable in solution (based on the information obtained from development report /
data), analysis may also be done at intermediate time intervals. However, report the results of initial and actual
time point.
Determine the absolute % difference in assay /response of standard peak and % impurity for sample solution at a
particular time point with respect to the initial % assay/response of standard peak and % impurity for sample
solution.

Acceptance criteria:
• The absolute difference in assay / response of standard peaks with respect to initial at each time
point shall not be more than 5.0. If it is outside the set criteria, make appropriate recommendations.
• The absolute difference of impurity results in the sample solution at each interval with respect to
initial shall not be more than 0.02.

7.2.6 FILTER COMPATIBILITY:

Determination
• If no known impurity is given in the specification, prepare sample solution as it is.
• If known impurity is given in the specification and is above LOQ level, prepare sample solution
as it is.
• If known impurity is given in specification and is below LOQ level, spike the impurity at the
specification limit in the sample solution.
Prepare the sample solution and placebo as described in the methodology. If multiple strength, select
• Higher strength, if linear dosages form.
• Lower Drug placebo ratio of dosage form, if average weight is same for all strength.

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 PHARMA DEVILS
QUALITY ASSURANCE DEPARTMENT

STANDARD OPERATING PROCEDURE

Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

At filtration stage, filter solution through Whatman GF/C filter, Whatman No.41, Whatman No.42; discard about
10 mL sample solution from each filter. For 0.45m membrane filter and 0.45µm PVDF discard about 5 mL
sample solution, collect the sample solutions for further analysis. Centrifuge the same (unfiltered) sample
solution. Analyse all the solution thus obtained and calculate the % impurity results. Determine the absolute
difference in the % impurity result obtained from filtered solution and centrifuged solution.
Each of the sample solutions thus obtained shall be analyzed as described in the methodology. The % impurities
shall be calculated. The absolute difference in the results obtained from filtered solutions and centrifuged solution
shall be calculated.

Acceptance criteria:
The absolute difference in % impurity result obtained from filter solution and centrifuge solution shall not be
more than 10 % of specification limit. If it is out of the set criteria, appropriate recommendation shall be made.

7.2.7 FILTER SATURATION:

Determination:
• If no known impurity is given in the specification, prepare sample solution as it is.
• If known impurity is given in the specification and is above LOQ level, prepare sample solution
as it is.
• If known impurity is given in specification and is below LOQ level, spike the impurity at the
specification limit in the sample solution.
Prepare the sample solution and placebo as described in the methodology. If multiple
strength, select
• Higher strength sample, if linear dosage forms.
• Lower Drug placebo ratio of dosage form, if average weight is same for all strengths.

At filtration stage, when Whatman filter paper no.41, Whatman filter paper no.42 and GF/C filter is recommended
for use in Filter compatibility study, filter 10.0 mL and 20.0 mL sample solution and discard using three separate
filters, followed by filtration of further 10 mL aliquots. Collect the filtrates each in separate test tubes. When
0.45µm nylon membrane filter and 0.45µm PVDF filter is recommended for use in Filter compatibility study
filter suitable volume of sample solution (1 ml,3 ml, & 5 ml) and discard using separate filters, followed by
filtration of further 10 mL aliquots. Collect the filtrates each in separate test tubes.

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Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

Analyse these solutions as described in the method to be validated and calculate the % impurity results. Determine
the absolute difference in the results obtained from two consecutive aliquots.
Acceptance criteria:
The absolute difference in % impurity result obtained in two consecutive filtrations shall not be more than 10 % of
specification limit. If it is out of the set criteria, make appropriate recommendations.

7.2.8 LINEARITY AND RANGE:

Determination:
Prepare linearity solutions from the stock solution of standards to obtain solutions at LOQ to 250% of the
specification limit by preparing minimum five concentration level. Inject each solution into the chromatograph in
single injection; calculate the peak area and corrected area.
Plot a graph of corrected areas vs. concentration (ppm) for each solution. Determine and report the slope,
intercept, correlation coefficient of the regression lines and residual sum of squares. For range, record the
concentration levels over which the results are linear.
Acceptance criteria:
Correlation Coefficient for each impurity shall be not less than 0.995.

7.2.9 PRECISION:
7.2.9.1 System precision:
Determination:
Prepare the standard solutions as described in the method to be validated and inject the obtained solutions in six
replicates. Calculate the relative standard deviation of the responses.
Acceptance criteria (For HPLC)
Relative standard deviation shall not be more than 5.0 % or as specified in the method.

7.2.9.2 Repeatability:
Determination:
• If no known impurity is given in the specification, prepare sample solution as it is.
• If known impurity is given in the specification and is above LOQ level, prepare sample solution
as it is.
• If known impurity is given in specification and is below LOQ level, spike the impurity at the
specification limit in the sample solution.
Repeatability shall be performed in the following way, for multiple strength.

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Department: Quality Assurance SOP No.:
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• If Sample is used for crushed powder for sample preparation, select only higher strength.
• If sample is used for intact tablets for sample preparation, all strengths to be performed.
• If sample dilution varies, repeatability shall be performed on all strengths.

Prepare six different sample solutions and placebo as described in the methodology and analyse using the method
to be validated, over a short period of time by same analyst, on same equipment, on same day. Calculate the %
impurity results. Determine the relative standard deviation and 95% confidence interval of the results obtained
from the six preparations.

Acceptance criteria:
Relative standard deviation of individual and total impurities results obtained in six preparations shall not be more
than 15.0 %.
7.2.9.3 Intermediate precision:
Determination:
• If no known impurity is given in the specification, prepare sample solution as it is.
• If known impurity is given in the specification and is above LOQ level, prepare sample
solution as it is.
• If known impurity is given in specification and is below LOQ level, spike the impurity at the
specification limit in the sample solution.

Prepare six different sample solutions and placebo as described in the Repeatability study and analyse using the
method to be validated, by different analyst, on different equipment, on different day. Calculate the % impurity
results. Determine the relative standard deviation and 95% confidence interval of the results obtained from the
six preparations. Only higher strength to be selected for intermediate precision.
Determine the relative standard deviation of the individual and total impurity results obtained from twelve
preparations of Repeatability and Intermediate precision.
Acceptance criteria:
• Relative standard deviation of individual and total impurities results obtained in six preparations
shall not be more than 15.0 %.
• Relative standard deviation of individual and total impurities results obtained in twelve
preparations (Repeatability and Intermediate precision) shall not be more than 15.0 %.
Note: The % Impurities below 0.05%; acceptance criteria shall not be applicable for it.

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Department: Quality Assurance SOP No.:
Title: Analytical Method Validation Effective Date:
Supersedes: Nil Review Date:
Issue Date: Page No.:

7.2.10 ACCURACY (RECOVERY):

Determination:
• If no known impurity is present, spike diluted standard at specification limit in placebo.
• For, known impurity spike the impurity in the sample solution at the specification limit.
• For, known impurities and unknown impurities, spike the known impurities and diluted standard
in placebo at the specification limit.

Prepare the test solution as described in methodology in triplicate and analyse. If multiple strength of different
colour, select worst condition of placebo which are,
• Lower Drug: placebo ratio if all strengths are linear.
• If different colour of material used for formulation and/ or coating, placebo shall be mixed with
all colours at proportionally.

Prepare the recovery solutions to obtain the solutions at LOQ to 250 % of specification limit by preparing
minimum three concentration level. Prepare the recovery solution at concentration levels in triplicate with single
injection and analyse the same. Calculate the mean areas of the peaks and the % results.
Acceptance criteria:
Recovery at LOQ to 250% shall be within 80.0 % to 120.0 %.

7.2.11 ROBUSTNESS:
Determination:
Prepare one sample solution and placebo as described in the Repeatability study and carry out analysis by making
individual, small, deliberate changes in the analytical procedure. Select the changes to be made in the analytical
procedure from the below list, as applicable.
• Change in pH of buffer (pH specified in method  0.2)
• Change in pH of mobile phase (pH specified in method  0.2)
• Change in mobile phase composition (Absolute 2 % or 30% of relative, whichever is larger).
• Change in flow rate (Flow rate specified in method  0.2)
• Change in column oven temperature (Temperature specified in method  5°C)
The result of % impurity shall be calculated for each set of analysis. The absolute difference in % impurity result
obtained in robustness study and method precision (sample-1) shall be calculated.

Acceptance criteria:

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The absolute difference in the results obtained in robustness study and Repeatability study (Sample-1) shall not be
more than 15 % of specification limit.

7.2.12 RELATIVE RESPONSE FACTOR:

Note: The relative response factor study may be carried out in the validation of an in-house related substances
method. The relative response factor allows for the quantitation of a known impurity against the diluted standard
of the drug substance, eliminates the need of using an impurity standard for every analysis. In the impurity
calculation formula, use the inverse of the relative response factor i.e. the correction factor.
Determination:
Calculate the relative response factor from the value of slope obtained in the linearity study using the
following formula:
Slope obtained for impurity
Relative Response Factor = -----------------------------------------------
Slope obtained for drug substance

Calculate the Correction Factor as follows:

1
Correction Factor = ------------------------------------
Relative Response Factor
Acceptance criteria:
Record the value of correction factor and use in the impurity calculation.

7.2.13 SYSTEM SUITABILITY:

Determination:
Perform System suitability before performing any parameter.
Acceptance criteria (For HPLC methods):
The system suitability shall comply as per methodology

7.3 ANALYTICAL METHOD VALIDATION FOR DISSOLUTION TEST OF DRUG

PRODUCT (IMMEDIATE RELEASE):
7.3.1 Parameters to be considered for an In-house method:
7.3.1.1 Specificity
7.3.1.2 Solution stability

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7.3.1.3 Filter compatibility

7.3.1.4 Filter saturation
7.3.1.5 Linearity and range
7.3.1.6 Precision (System precision, Repeatability, Intermediate precision)
7.3.1.7 Accuracy
7.3.1.8 Robustness
7.3.1.9 System suitability

7.3.2 Parameters to be considered for a Pharmacopoeial method:

7.3.2.1 Specificity
7.3.2.2 Solution stability
7.3.2.3 Filter compatibility
7.3.2.4 Filter saturation
7.3.2.5 Precision (System precision, Repeatability, Intermediate precision)

7.3.3 SPECIFICITY:
Determination:
Prepare a blank solution (diluent), placebo solution, standard solution and sample solution and
analyse to check for interference.

Acceptance criteria:
For HPLC method:
No peak shall be observed due to blank solution and placebo solution at the retention time of
principle peak as observed in the standard solution and sample solution.
For UV method:
Interference due to placebo solution shall not be more than 2.0 %.

7.3.4 SOLUTION STABILITY:

Determination:
Prepare standard solution and sample solution (equivalent to target concentration) as described in the method to
be validated. Keep the prepared solutions tightly closed and store at room temperature and at 10°C. Analyse the
standard and sample solution up to 8 hours (if estimation method is by UV spectrophotometry) or analyse the
standard and sample solution up to 24 hours of both the condition (if possible 48 h) (if estimation methods is

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HPLC) if the drug product tends to be stable in solution (based on the information obtained from development
report / data), analysis may also be done at intermediate time intervals. However, report the results of the initial
and actual time point. Determine the absolute % difference in assay at the particular time point the initial assay for
standard and % Dissolution release in test solution.
Acceptance criteria (For HPLC & UV methods):
The absolute % difference in assay/ % release with respect to initial at each time point shall not be
more than 2.0. If it is out of the set criteria, make appropriate recommendations.

7.3.5 FILTER COMPATIBILITY:

Determination:
Prepare the sample solution (equivalent to target concentration) as described in the method to be validated. If
multiple strength, select higher strength if dosage formula is linear.
At the filtration stage, filter the sample solution through Whatman GF/C filter, Whatman No.41,
Whatman No.42; discard about 10 mL sample solution from each filter. For 0.45m membrane filter
and 0.45µm PVDF discard about 5 mL sample solution, collect the sample solutions for further
analysis. Centrifuge the same (unfiltered) sample solution. Analyse all the solutions and calculate
the results. Determine the absolute difference in the results obtained for the filtered solution and
centrifuged solution.
Acceptance criteria (For HPLC & UV methods):
The absolute difference in results obtained for the filtered solutions and the centrifuged solution shall
not be more than 2.0. If it is out of the set criteria, make appropriate recommendations.
7.3.6 FILTER SATURATION:
Determination:
Prepare the sample solution (equivalent to target concentration) as described in the method to be validated. If
multiple strength, select higher strength if dosage formula is linear. At filtration stage, when Whatman filter paper
no.41, Whatman filter paper no.42 and GF/C filter is recommended for use in Filter compatibility study, filter
10.0 mL and 20.0 mL sample solution and discard using three separate filters, followed by filtration of further 10
mL aliquots. Collect the filtrates each in separate test tubes. When 0.45µm nylon membrane filter and 0.45µm
PVDF filter is recommended for use in Filter compatibility study filter suitable volume (1mL, 3mL and 5mL)
of sample solution and discard using separate filters, followed by filtration of further 10 mL aliquots. Collect the
filtrates each in separate test tubes.
Analyse the thus obtained solutions as described in the method to be validated and calculate the results. Determine
the absolute difference in the results obtained for two consecutive aliquots.

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Acceptance criteria (For HPLC & UV):

The absolute difference in the result obtained for two consecutive filtered aliquots shall not be more than 2.0. If it
is out of the set criteria, make appropriate recommendation.

7.3.7 LINEARITY AND RANGE:

Determination:
Prepare linearity solutions from the stock solution of standard to obtain the solutions at 30 % to 150 % of the
working concentration by preparing minimum 5 concentration level. In case of multiple strength of dosage form,
shall cover from 30 % of lower strength and up to 150 % of higher strength by preparing minimum 5
concentration level. Inject all the prepared solutions in single injection.
Plot a graph of corrected area vs. concentration (ppm). Determine and report the slope, intercept, and correlation
coefficient of the regression line and residual sum of squares. For range, record the concentration levels over
which the results are linear.
Acceptance criteria (For HPLC & UV methods):
Correlation Coefficient shall be not less than 0.995

7.3.8 PRECISION:
7.3.8.1 System precision:
Prepare the standard solution as described in the method to be validated and inject the obtained solution in six
replicates. Calculate the relative standard deviation of the responses.
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation shall not be more than 2.0 %.
7.3.8.2 Repeatability:
Determination:
Perform the dissolution test for six dosage units using the method under validation and analyzed by same
analyst, on the same equipment, on same day. For multiple strength, repeatability, perform all the strengths and
calculate the dissolution results. Determine the relative standard deviation and 95 % Confidence interval of the
dissolution results (six dosage units).
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation of dissolution results (six dosage units) shall not be more than 5.0 %
7.3.8.3 Intermediate precision:
Determination:

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Perform the dissolution test on six dosage units using the method under validation and analyzed by a different
analyst, on different equipment, on different day. Calculate the dissolution results. Determine the relative standard
deviation and 95 % Confidence interval of the dissolution results (six dosage units). Only higher strength to be
selected for intermediate precision.
Calculate the absolute difference in the results obtained in Repeatability (mean value of six dosage units) and
Intermediate precision (mean value of six dosage units).
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation of dissolution results (six dosage unit) shall not be more than 5.0 %. The absolute
difference in the dissolution results obtained in Repeatability (Mean result of six dosage units) and Intermediate
precision (Mean result of six dosage units) shall not be more than 5.0 %.

7.3.9 ACCURACY (RECOVERY):

Determination:
Prepare recovery solutions by spiking the drug substance in to the dissolution vessel containing placebo powder
equivalent to one dosage unit to cover from 50 % to 120 % of target concentration by preparing minimum 3
concentration level.
If multiple strength select
• Higher strength if dosage formula is linear. If the formula is linear and coating material are of
different colors use placebo mixed with all the color proportionally.
• Lower drug placebo ratio of dosage form if average weight is same for all strengths.
Analyse the solutions as described in the method under validation.
If the amount of drug substance to be spiked to each dissolution vessel is less than 10 mg or if the drug
substance floats in the dissolution medium or if the drug substance is poorly soluble, then prepare a stock solution
of drug substance for spiking into the dissolution vessels.
Calculate the recovery in mg and as % recovery for each level and mean recovery of all six solutions.
Acceptance criteria (For HPLC & UV methods):

All the individual recoveries shall be within 95.0 % to 105.0 %.

The relative standard deviation for % recovery shall not be more than 5.0 %.

7.3.10 ROBUSTNESS:
Determination:

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Carry out one set (dissolution test of six dosage units) of analysis using the same batch of the product, by making
individual, small, deliberate changes in the analytical procedure. Select the changes to be made in the analytical
procedure from the below list, as applicable.
• Change in medium volume ( 1%).
• Change in pH of dissolution medium (pH specified in dissolution medium  0.2).
• Change in strength of the dissolution medium (specified morality  0.02 M).
• Change in pH of mobile phase (pH specified in method  0. 2).
• Change in mobile phase composition (Absolute 2% or 30% of relative, whichever is larger).
Calculate the dissolution results for each set of analysis. Determine the absolute difference between the results
obtained in Robustness study (Mean dissolution) and Repeatability study (Mean dissolution).
Acceptance criteria (For HPLC & UV methods):
The absolute difference in the results obtained in Robustness study (Mean dissolution) and Repeatability study
(Mean dissolution) shall not be more than 5.0%.
7.3.11 SYSTEM SUITABILITY
Determination:
Perform System suitability before performing any parameter.
Acceptance criteria (For HPLC & UV methods):
The system suitability shall comply as per methodology.

7.4 ANALYTICAL METHOD VALIDATION FOR DISSOLUTION TEST OF DRUG

PRODUCT (MODIFIED RELEASE).
7.4.1 Parameters to be considered for an In-house method:
7.4.1.1 Specificity
7.4.1.2 Solution stability
7.4.1.3 Filter compatibility
7.4.1.4 Filter saturation
7.4.1.5 Linearity and range
7.4.1.6 Precision (System precision, Repeatability, Intermediate precision)
7.4.1.7 Accuracy
7.4.1.8 Robustness
7.4.1.9 System suitability

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7.4.2 Parameters to be considered for a Pharmacopoeial method:

7.4.2.1 Specificity
7.4.2.2 Solution stability
7.4.2.3 Filter compatibility
7.4.2.4 Filter saturation
7.4.2.5 Precision (System precision, Repeatability, Intermediate precision)
7.4.3 SPECIFICITY:
Determination:
Prepare a blank solution (diluent), placebo solution, standard solution and sample solution and analyse to check
for interference.
Use placebo in the finished dosage form and inject at each interval
Acceptance criteria:
For HPLC method:
No peak shall be observed due to blank solution and placebo solution at the retention time of principle peak as
observed in the standard solution and sample solution.
For UV method:
Interference due to placebo solution shall not be more than 2.0 %.

7.4.4 SOLUTION STABILITY:

Determination:
Prepare Standard solution and sample solution (at 100 % level) as described in method under validation. Keep the
prepared solutions tightly closed and store at room temperature, 10°C and at 37°C and analyse at the intervals
specified in methodology and at 6 hours beyond the last time point of all condition. Determine the absolute %
difference in assay/% dissolution release at each time point with respect to the initial assay/% dissolution release
for standard and test solutions
Acceptance criteria (for HPLC & UV methods):
The absolute difference in assay, with respect to initial, at each time point shall not be more than 2.0. If it is out of
the set criteria, make appropriate recommendations

7.4.5 FILTER COMPATIBILITY:

Determination:

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Prepare the sample solution at concentration equivalent to the limit specified in methodology at the first time point
and last time point. At the filtration stage, filter the solution through Whatman GF/C filter, Whatman No.41,
Whatman No.42; discard about 10 mL sample solution from each filter. For 0.45m membrane filter and 0.45µm
PVDF discard about 5 mL sample solution, collect the sample solutions for further analysis. Centrifuge the same
(unfiltered) sample solution. Analyse all the solutions and calculate the results. Determine the absolute difference
in the results obtained for the filtered solutions and the centrifuged solutions.
Acceptance criteria (For HPLC & UV methods):
The absolute difference in result obtained from filtered solution and centrifuged solution shall not be more than
2.0. If it is out of the set criteria, make appropriate recommendations.

7.4.6 FILTER SATURATION:

Determination:
Prepare the sample solution at concentration equivalent to limit specified in methodology for the first time point
and last time point. At filtration stage, when Whatman filter paper no. 41, Whatman filter paper no.42 and GF/C
filter is recommended for use in Filter compatibility study filter 10.0 mL and 20.0 mL sample solution and discard
using three separate filters, followed by filtration of further 10 mL aliquots. Collect the filtrates each in separate
test tubes. When 0.45µm nylon membrane filter and 0.45µm PVDF filter is recommended for use in Filter
compatibility study filter suitable volume of sample solution (1 ml, 3 ml & 5ml) and discard using separate
filters, followed by filtration of further 10 mL aliquots. Collect the filtrates each in separate test tubes. Analyse
the thus obtained solutions as described in the method under validation and calculate the results. Determine the
absolute difference in the results obtained for two consecutive aliquots.
Acceptance criteria (For HPLC & UV methods):
The absolute difference in result obtained in two consecutive filtrations shall not be more than 2.0. If it is out of
the set criteria, make appropriate recommendations.
7.4.7 LINEARITY AND RANGE:
Determination:
Prepare linearity solutions from the stock solution of standard to obtain the solutions at 30% of limit specified at
first time point to 150% of the limit specified at last time point. This range must contain minimum 9 linearity
points. Inject all the prepared solutions in single injection.
Plot a graph of corrected area vs. concentration (ppm). Determine and report the slope, intercept, and correlation
coefficient of the regression line and residual sum of squares. For range, record the concentration levels over
which the results are linear.

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Acceptance criteria (For HPLC & UV methods):

Correlation Coefficient shall be not less than 0.995

7.4.8 PRECISION:
7.4.8.1 System precision:
Prepare the standard solution as described in the method to be validated and inject the obtained solution in six
replicates. Calculate the relative standard deviation of the responses.
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation shall not be more than 2.0 %.
7.4.8.2 Repeatability:
Determination:
Perform the dissolution test for six dosage units using the method under validation and analyzed by same analyst,
on the same equipment, on the same day. For multiple strength, precision shall carried out at all the strengths and
Calculate the dissolution results at each time point. Determine the relative standard deviation and 95% Confidence
interval of the dissolution results (six dosage units) for each time point.
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation of dissolution results (six dosage unit) at each time point shall not be more than 10.0
%.
7.4.8.3 Intermediate precision:
Determination:
Perform the dissolution test on six dosage units using the method under validation and analyzed by a different
analyst, on different equipment, on different day. For multiple strength only higher strength shall be selected and
Calculate the dissolution results for each time point. Determine the relative standard deviation of the dissolution
results (six dosage units) for each time point.
Calculate the absolute difference in the results obtained in Repeatability (mean value of six dosage units) and
Intermediate precision (mean value of six dosage units) for each time point.
Acceptance criteria (For HPLC & UV methods):
Relative standard deviation of dissolution results (six dosage unit) at each time point shall not be more than 10.0
%.
The absolute difference in the dissolution results at each time point obtained in Method precision (Mean results of
six dosage units) and Intermediate precision (Mean results of six dosage units) shall not be more than 10.0 %.

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7.4.9 ACCURACY (Recovery):

Determination:
Prepare recovery solutions by spiking the drug substance in to the dissolution vessel containing placebo powder
equivalent to one dosage unit to obtain the solutions at 50% of limit specified at first time point, 100% of limit
specified at last time point and 120% of limit specified at last time point, in triplicate.
Analyse the solutions as described in the method under validation.
If the amount of drug substance to be spiked to each dissolution vessel is less than 10 mg or if the drug substance
floats in the dissolution medium or if the drug substance is poorly soluble, then prepare a stock solution of drug
substance for spiking into the dissolution vessels.
Calculate the recovery in mg and as % recovery for each level and mean recovery of all solutions.
Acceptance criteria (For HPLC & UV methods):
All the individual recoveries shall be within 95.0 % to 105.0 %.
The relative standard deviation for % recovery shall not be more than 5.0 %.

7.4.10 ROBUSTNESS:
Determination:
Carry out one set (dissolution test of six dosage units) of analysis using the same batch of the product, by making
individual, small, deliberate changes in the analytical procedure. Select the changes to be made in the analytical
procedure from the below list, as applicable.
• Change in medium volume (1 %).
• Change in pH of mobile phase (pH specified in mobile phase  0.2)
• Change in strength of the dissolution medium (specified molarity  0.02 M)
• Change in mobile phase composition (Absolute 2% or 30% relative, whichever is larger.)
Calculate the dissolution results for each set of analysis. Determine the absolute difference between the results
obtained at each time point in Robustness study (mean dissolution) and Repeatability study (mean dissolution).
Acceptance criteria (For HPLC & UV methods):
The absolute difference in the results obtained in Robustness study (mean dissolution) and Repeatability study
(Mean dissolution) at each time point shall not be more than 10.0.

7.5 ANALYTICAL METHOD VALIDATION FOR RESIDUAL SOLVENTS TEST OF DRUG

PRODUCT.
Parameters to be considered for an in House method.

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• Specificity
• Linearity and range
• Precision (System precision, Method precision, Ruggedness)
• Accuracy
• Robustness
• System suitability

7.5.1 SPECIFICITY
7.5.1.1 Interference study
Determination:
Drug products: Prepare the blank (diluent), individual known residual solvents, sample solution and six injections of
standard as described in the methodology and inject into the GC system. Record the retention time (RT) of All peaks
observed in the resulting chromatograms. Inject placebo to check the interference.
Acceptance criteria:
No interference of solvents from each other at the retention time of analyte.

7.5.2 LIMIT OF DETECTION & LIMIT OF QUANTITATION:

Prepare a series of equivalent lowest concentration solutions by quantitative dilutions of the standard stock solution
of residual solvents. Inject each solution and record the peak areas.
Determine the slope and standard deviation of the response by plotting a graph of peak area vs. concentration (ppm).
Determine the value of LOD and LOQ using the following formula:
Calculation:

3.3 x STEYX 10 x STEYX

LOD = ------------------- LOQ = --------------------
Slope Slope

Where,
STEYX = Residual sum of squares
LOD = Limit of detection
LOQ = Limit of quantitation

Prepare the solution at LOQ level and inject six injections and calculate the relative standard deviation of the peak

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areas.
Acceptance criteria:
% RSD of the peak area of residual solvents in six injections of LOQ and LOD is not more than 15.0 and 33.0
respectively.

7.5.3 LINEARITY AND RANGE:

Determination:
Prepare linearity solutions by quantitative dilutions of the stock solution of standards to obtain solutions at LOQ and
minimum five levels between 50 % and 150 % level of the specification limit and inject each solution into the gas
chromatogram in triplicate and calculate the mean peak area of each solvents.
Plot a graph of mean peak areas vs. concentration (ppm) and determine the equation of regression line. Report
the slope, intercept and correlation coefficient of the regression lines and residual sum of squares. For range, record
the concentration levels over which the results are linear.
Acceptance criteria:
Correlation Coefficient shall not be less than 0.995.
7.5.4 PRECISION
7.5.4.1 System precision:
To check the system precision, prepare the standard solution as per the methodology and record the peak areas of
six injections of standard solution. Calculate the mean, standard deviation and % RSD of each residual solvent.
Acceptance criteria:
System suitability criteria shall meet
7.5.4.2 Repeatability:
Determination:
If any analyte is present in the sample prepare six-test solution.
if any analyte is not present in the sample, prepare six test solutions followed by six test solutions by spiking with
standard solution at 100 % level of the specification limit.
Calculate the mean, standard deviation and % RSD of residual solvent results.
Acceptance criteria :
The % RSD for each residual solvent results in method precision is not more than 15.0.
7.5.4.3 Ruggedness:
Determination:
If any analyte is present in the sample prepare six-test solution.

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If any analyte is not present in the sample, prepare six test solutions followed by that six test solutions by spiking
with standard solution at 100 % level of the specification limit.
Calculate the mean, standard deviation and % RSD of residual solvent results.
Acceptance criteria:
The overall % RSD of the residual solvent results obtained from method precision and ruggedness is not more than
15.0.

7.5.5 ACCURACY (Recovery)

Determination:
Prepare three test solution as per the methodology and record the areas.
Spike the test sample with standard solution, at three levels in the range of 50%, 100% and 150% of specification
limit.
Calculate mean recovery, standard deviation and the % RSD of recovery results.
Acceptance criteria:
The % Recovery of each residual solvent at 50%, 100% and 150% levels of the specification limit shall be within the
range 80 % to 120%.

7.5.6 ROBUSTNESS:
Determination:
Carry out GC analysis as per method precision by making the following changes in the chromatographic conditions
(±10%).
If any analyte is present in the sample prepare two test solutions.
If any analyte is not present in the sample, prepare two test solutions by spiking with standard solution at 100% level
of the specification limit.
• Change the flow rate of carrier gas (Flow rate specified in method ± 10% variation).
• Change the temperature of the column oven (initial column oven temperature specified in method ±
10% variation).
• Change the Headspace Equilibration temperature. (Equilibration temperature specified in the
method ± 10% variation).
Acceptance criteria:
Shall meet all the requirement of system suitability

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7.6 Documents and reporting of results of analytical method validation

7.6.1 All method validation activities shall be recorded on the raw data sheet.
7.6.2 After completion of analysis, the analyst shall sign in the Record of analysis and submit the same to the
Head Q.C or designee.
7.6.3 Head Q.C his designee shall check all documentation, calculations and sign in the Record of analysis in
the checked by column.

7.7 Method validation report.

7.7.1 Method Validation protocol shall be prepared for each individual test /product before the method
validation study to be started.
7.7.2 Method Validation report shall be prepared once the analytical activity of method validation study is
completed, all the relevant raw data are verified, and all the calculations are checked.
7.7.3 A summary shall be mentioned of method validation report followed by recommendation if any.
7.7.4 Method Validation report shall clearly identify the critical parameter, shall suggest the
precautions to be taken care while actual use of methodology and shall suggest any modifications to be
done in methodology.
7.7.5 Method Validation report shall be checked, reviewed and approved by the respective authorities.
7.7.6 Original copy of method validation protocol and validation report shall be filed together with proper
labeling along with all the relevant raw data.

8.0 REVISION HISTORY

Version No. 00 Effective Date
Details of revision: New SOP Prepared

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