Analytical Method and Processes Validation

Presented by Yub Raj Neupane

TOPICS.
INTRODUCTION....... TYPES OF ANALYTICAL PROCEDURES ..... REFERENCE STANDARDS ...... METHODS VALIDATION FOR INDS .. VALIDATION CHARACTERISTICS........ METHODOLOGY....

INTRODUCTION.......
Process of demonstrating that analytical procedures are suitable for their intended use. FDA, NDA or ANDA applicant must submit samples of drug product , drug substance, non compendial reference standards with analytical procedures.

Steps in Method Validation

Protocol or operating procedure Application, purpose, and scope of the method. Performance parameters and acceptance criteria. Qualify materials (e.g., standards and reagents). Criteria for revalidation. Document validation experiments

TYPES OF ANALYTICAL PROCEDURES

A. Regulatory Analytical Procedure: To evaluate a defined characteristic of the drug substance or drug product. In the U.S. Pharmacopeia/National Formulary (USP/NF).

B. Alternative Analytical Procedure: Proposed by the applicant for use instead of the regulatory analytical procedure.

C. Stability-Indicating Assay:

Can detect the changes with time in the pertinent properties of the drug substance and drug product.

REFERENCE STANDARDS
Reference standard Obtained from the USP/NF or other official sources Working standard: Is qualified against and used instead of the reference standard.

METHODS VALIDATION FOR INDs

Investigational new drug, sufficient information is required in each phase of an investigation to ensure proper identification, quality, purity, strength, and/or potency. Guidance on analytical procedures and methods validation information to be submitted for different phases of studies, sponsors should refer to the FDA guidance for industry.

PARAMETERS FOR METHOD VALIDATION

SELECTIVITY AND SPECIFICITY

Specific: -Method that produces a response for a single analyte only Selectivity: measure accurately an analyte in the presence of interference ( precursors, excipients, enantiomers) and degradation products that may be expected to be. In LC selectivity is obtained by choosing optimal columns and setting chromatographic conditions, such as mobile phase composition, column temperature, and detector wavelength

PRECISION AND REPRODUCIBILITY

Repeatability: In one laboratory by one operator using one piece of equipment over a relatively short time span. Precision of better than 1% RSD is easily achieved. Intermediate precision: Comparing the results of a method run within a single laboratory over a number of weeks.

 Reproducibility:
Represents the precision obtained between laboratories.

Important if the method will be used in different laboratories

ACCURACY AND RECOVERY

Test results generated by the method and the true value agree. Compare the results of the method with results from an established reference method. By analyzing a sample with known concentrations.

LINEARITY AND CALIBRATION CURVE

Test results that are directly, or by means of well-defined mathematical transformation, proportional to the concentration of analytes in samples within a given range. A linear regression equation applied to the results should have an intercept not significantly different from zero.

RANGE
Interval between the upper and lower levels. Be determined with precision, accuracy, and linearity using the method as written

LIMIT OF DETECTION AND QUANTITATION

Lowest concentration of an analyte in a sample that can be detected but not necessarily quantified. In chromatography LOD is the injected amount that results in a peak with a height at least twice or three times as high as the baseline noise level LOQ is the minimum injected amount that gives precise measurements, in chromatography typically requiring peak heights 10 to 20 times higher than baseline noise.

ROBUSTNESS
Examine the effect operational parameters have on the analysis results.

Flow rate, column temperature, injection volume, detection wavelength, or mobile phase composition.

METHODOLOGY
High-Pressure Liquid Chromatography (HPLC): Column: -glass, stainless steel, plastic - length, inner diameter -Particle type of packing materials System Suitability Testing -Relative retention -Relative standard deviation (RSD)

 Gas Chromatography GC: Column: - length, internal diameter, external diameter. -Stationary phase Gases: purity, flow rate, pressure Temperatures: column, injector, detector Injection .e.g., split, splitless, on-column Detector Typical retention time and total run time

 Capillary Electrophoresis (CE): Capillary: - length, length to detector, internal diameter, external diameter -Capillary material - Running buffer - Detector - Voltage -Current