9 Most Important Parameter for Analytical method validation

Analytical method validation parameters are following describe:

1)   Selectivity/specificity

Selectivity is measured accurately by an analyte in the presence of interferences that might be expected to be present in the sample matrix. It is checked by examining chromatographic blanks in the anticipated time window of the analyte peak.

2)   Precision

Precision is the degree of agreement between individual test results after the procedure is applied repeatedly to multiple samplings under similar conditions. It is measured by injecting a series of standards or a series of analyzing samples from multiple samplings from a homogeneous batch of material. It aims to demonstrate the random error that could occur in a method. 

3)   Accuracy

"Accuracy is that the degree of agreement among the value which is accepted whichever as a conventional true value or an accepted reference value and therefore the value found". It is measured with a known concentration of analyte standard and analyzing the sample by spiking the sample matrix of interest.

4)   Linearity

Linearity is expressed by injecting a series of standards of stock solution or diluted stock solution using the solvent or mobile phase, at a minimum of 5 different concentrations in the range of 50–150% of the anticipated working range. It is simple between Concentrations vs. Peak Area Response.

5)   Range

The range (the interval between the upper and lower levels) of an analytical method has been demonstrated to determine precision, accuracy, and linearity using the set method. This range is defined by the concentration range in which the Linearity test was completed.

6)   Stability

Many samples decompose during chromatography studies, such as in the preparation of the sample solutions, phase transfer, clean-up, extraction, and storage of prepared vials. Accuracy is takings care of stability, how long a sample solution can be stored before final analysis.

7)   LOD and LOQ

Limit of detection (LOD) is defined as the lowest concentration at which point the instrument can identify but not quantify, and the noise/signal ratio for LOD must be 1:3.

Limit of quantitation (LOQ) is defined as the lowest concentration at which point the instrument can detect and quantify, and the noise/signal ratio for LOQ should be 1:10.0

8) Robustness (or Ruggedness)

The procedure to provide analytical results of acceptable accuracy and precision under a variety of conditions such as reagents (e.g. different suppliers), different columns (e.g. different lots and/or suppliers), extraction time, Variations of pH of a mobile phase, variations in mobile phase composition, temperature, and flow rate.

9) Forced degradation

These studies are done to degrade the active drug deliberately. The drug substance is exposed to thermal, photometric, acid, base, oxidizing agent, reducing agent, and water. These studies are used to estimate an analytical method’s ability to measure an active ingredient substance and its degradation products without any interference.

- Pritam Surkar