EffiValidation 3.0 is a compliance-ready ISO 17025 and GxP laboratory software.

Method Validation


ISO 17025:2005 definition

Validation is the confirmation by examination and the provision of effective evidence that the particular requirements for a specific intended use are fulfilled.

ICH Q2(R1) definitions


Specificity

Specificity is the ability to assess unequivocally the analyte in the presence of components which may be expected to be present. Typically these might include impurities, degradants, matrix, etc. Lack of specificity of an individual analytical procedure may be compensated by other supporting analytical procedure(s).

EffiValidation 3.0: Example of a Selectivity determination by Comparison of Calibration Lines

Method validation - Selectivity.jpg

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 value found. This is sometimes termed trueness.

EffiValidation 3.0: Example of the Accuracy determination by a t-test

Method validation - Accuracy - t-test.jpg

Precision

The precision of an analytical procedure expresses the closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions. Precision may be considered at three levels: repeatability, intermediate precision and reproducibility. Precision should be investigated using homogeneous, authentic samples. However, if it is not possible to obtain a homogeneous sample it may be investigated using artificially prepared samples or a sample solution. The precision of an analytical procedure is usually expressed as the variance, standard deviation or coefficient of variation of a series of measurements.

(i) Repeatability

Repeatability expresses the precision under the same operating conditions over a short interval of time. Repeatability is also termed intra-assay precision.

EffiValidation 3.0: Example of the Repeatability determination Level by Level from Multiple Measurements

Methon Validation - Precision as Repeatability (Level by Level from Multiple Measurements).jpg

(ii) Intermediate precision

Intermediate precision expresses within-laboratories variations: different days, different analysts, different equipment, etc.

(iii) Reproducibility

Reproducibility expresses the precision between laboratories (collaborative studies, usually applied to standardization of methodology).

Detection Limit

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.

Quantitation Limit

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. The quantitation limit is a parameter of quantitative assays for low levels of compounds in sample matrices, and is used particularly for the determination of impurities and/or degradation products.

EffiValidation 3.0: Example of the Detection and Quantitation Limit determination from a calibration curve

Method validation - Limit of detection_quantitation.jpg

Linearity

The linearity of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample.

EffiValidation 3.0: Example of the Linearity determination

Method validation - Linearity.jpg

Range

The range of an analytical procedure is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity.

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.

EffiValidation 3.0: Example of the Robustness determination

Method validation - Robustness_Ruggedness.jpg