Analytical method validation
Purpose
1.1 To describe a procedure for the validation of analytical methods.
1.0 Scope
2.1 This standard operating procedure is applicable for validation of analytical methods for Drug
products in the QC laboratory at ABC Pvt. Ltd., Roorkee (UK), India.
2.0 Responsibility
3.1 Officer QC/ Executive QC Preparation of SOP and follow the procedure as per SOP
Assistant Manager Reviewing and Training of SOP
Manager QC Implementation of SOP
3.0 Accountability
3.1 Manager Quality Control is accountable for implementation and compliance with SOP.
4.0 Procedure
4.1 Analytical methods used for the determination of assay, determination of related substances shall be validated.
4.2 Analytical methods shall be validated for the following parameters.
4.3 For assay of Active Pharmaceutical Ingredients (API) & Drug Products
4.3.1.1 Linearity
4.3.1.2 Accuracy
4.3.1.3 Precision– System, Repeatability & Intermediate Precision
4.3.1.4 Specificity
4.3.1.5 Robustness
4.3.2 For related substances of Active Pharmaceutical Ingredients (API) & Drug Products
4.3.2.1 Linearity
4.3.2.2 Accuracy
4.3.2.3 Precision – System, Repeatability & Intermediate Precision
4.3.2.4 Specificity
4.3.2.5 Limit of Detection
4.3.2.6 Limit of Quantification
4.3.2.7 Robustness
4.3.3 For Dissolution and drug release of the Drug products:
4.3.3.1 Linearity
4.3.3.2 Specificity
4.3.3.3 Precision – System, Repeatability & Intermediate Precision
4.3.3.4 Robustness
4.3.4 Revalidation shall be done if there is change in Master formula of drug product,
or any other change in the analytical method.
4.3.5 The ‘Analytical Method Validation Protocol’ shall address the following parameters:
4.3.6 Linearity
4.3.6.1 The linearity of an analytical method is its ability to elicit test results that are directly,
or by a well defined mathematical transformation proportional to the concentration
of analyte in sample in given range.
4.3.6.2 Linearity shall be established across the range above and below at minimum five
concentration level, of the analytical procedure. The linearity shall be established by the
examination of plot of signal verses the concentration of analyte. The plot should be a
linear line with regression co-efficient not less than 0.99.
4.3.7 Precision
4.3.7.1 Precision of the analytical is the degree of agreement among the individual test results
when the method is applied repeatedly to a multiple sampling of a homogeneous sample.
The precision shall be expressed as the related standard deviation of a series of measurements.
4.3.8 System Precision
4.3.8.1 The system precision shall be established for reproducible performance of instruments.
4.3.9 Repeatability
4.3.9.1 Make 09 independent sample solutions of a pooled sample at three different
levels (50%, 100% and 150%) and analyse by the method. Calculate the assay for each analysis and mean & RSD.
4.3.10 Intermediate Precision (Ruggedness)
4.3.10.1 Analysis of a 03 independent sample solutions (50%, 100% and 150%) of a pooled
sample by this method in triplicate by two different Analysts on different days. Calculate the
assay for each analysis. Calculate the mean and RSD.
4.3.11 Accuracy
4.3.11.1 The accuracy of an analytical method is the closeness of test results obtained by that
method to the true value. Accuracy shall be calculated as the percentage of recovery by the
assay of known added amount of analyte in the sample. The accuracy of method shall be
accessed by minimum of nine determinations over minimum of three concentration levels,
covering the specified range (i.e. three concentrations and three replicates of each concentration.
4.3.12 Specificity
4.3.12.1 Specificity is defined as the ability to access unequivocally the analyte in the
presence of components that may be expected present, such as impurities,
degradation products and matrix components.
4.3.12.2 The specificity of an analytical method is determined by comparing test results
from the analysis of samples containing impurities, degradation products, or placebo
ingredients with those obtained from the analysis of samples without impurities,
degradation products, or placebo ingredients. When impurities or degradation
products are unidentified or unavailable, specificity may be demonstrated by analysis
by the method in question of samples containing impurities or degradation product
and comparing the results to those from additional purity assay. Specificity of an analytical
method shall not be established if it is already established in analytical Development department.
4.3.13 Limit of Detection
4.3.13.1 The determination of limit of detection of an analytical method shall depend on
the method i.e. instrumental or instrumental procedure. For instrumental procedure
Signal to noise ratio shall be measured. The signal to ration 3:1 shall be considered as the limit of detection.
4.3.13.2 For non-instrumental methods limit of detection is generally determined by
analysis of samples with known concentration of analyte and by establishing a minimum
level at which the analyte can be reliably detected.
4.3.14 Limit of Quantification
4.3.14.1 The determination of limit of quantification of an analytical method shall vary
with the method i.e. instrumental or non-instrumental procedure. For instrumental
procedure measure the magnitude of analytical background response by analysing No.
of blank samples and calculating the standard deviation of the response. The standard
deviation multiplied by the factor 10 provides an estimate of limit of quantification.
The limit is subsequently validating by analyzing a suitable no. of samples known to be
near or prepared at the limit of quantification.
4.3.14.2 For non instrumental methods limit of quantification is generally determined by
analysis of samples with known concentration of analyte and by establishing a minimum
level at which the analyte can be reliably detected with acceptable accuracy and precision.
4.3.15 Robustness
4.3.15.1 As appropriate, make following changes in the experimental conditions — Variation
in mobile phase composition or flow rate.
4.3.15.2 Under each of the above variation of mobile phase composition or flow rate, perform
analysis of sample. Calculate the assay, and cumulative RSD of each condition with precision results.
5.0 Reference
ICH Guideline Q2 R2/ USP
6.0 Attachments
Not Applicable
7.0 Distribution
Distribution of the controlled copy to concerned departments.
Additional copies may be issued for specific purpose.
8.0 Abbreviations
QA : Quality Assurance
QC : Quality Control
SOP : Standard Operating Procedure
9.0 Revision History
| Revision No. | Supersedes | Effective Date | Details of Revision |
| 01 | Nil | New SOP |
Operating Procedure of Centrifuge
sop for preparation and storage of reagent solution
sop for Calibration and Maintenance of Laboratory Instruments and Equipment
Disposal of Residual Sample or Left Over Material
sop for for Laboratory Incident
standard operating procedure temperature monitoring
sop for operation of infrared moisture balance
sop for preparation of mobile phase
sop for Preparation and Issuance of Analysis protocol standard
sop of placebo and impurity stock solutions
sop for disposal of residual sample
sop for handling of pharmacopoeial changes
sop for procedure for operation of ultrasonic cleaner
difference between UPLC and HPLC
sop for for Emergency Eyewash and Shower
sop for operation and calibration of total organic carbon analyzers
sop for operation of cobb tester
sop for Operation and calibration of atomic absorption spectrophotometer
sop for Operation and calibration of gas liquid chromatograph
sop for operation of humidity oven
sop for operation and calibration of serological water bath
sop for monitoring of drain trap
sop for destruction of analytical samples after testing and control samples
sop for destruction of used chemicals
Sop for Operation of suction pump
sop for Operation and calibration uv cabinet
sop for Operation and calibration of bulk density apparatus
sop for operation and calibration of shore hardness tester
sop for operation of rub proofness tester
sop for monitoring of purified water
sop for Retesting of packaging materials
sop for Retesting and resampling of raw materials
sop for Control of issuance of record of analysis green sheets
sop for Control of computer passwords
sop for sampling of packaging materials PM
Operation and Calibration Procedure for pH Meter
operation and calibration of muffle furnace
Operation and cleaning of centrifuge
Operation and cleaning of sonicator
Operation and calibration of dissolution test apparatus
Operation and cleaning of hot air oven
Operation and calibration of bursting strength
Operating procedure of refrigerator
Operating procedure of stop watch
