Bioanalytical Methods Validation for Human Studies - Pharmanet

Draft — Not for Implementation
for the development of SOPs for evaluation of an analyte’s stability in a biological
matrix (Timm 1985). Whatever approach is used, the SOPs should clearly
describe the statistical method and rules employed. Additional validation may
include investigation of samples from dosed subjects.
F. Acceptance Criteria
An analytical method is considered fully validated when it meets the following criteria:
Precision: The between-batch CVs for low, medium, and high concentrations should be
#15%, and #20% for the LOQ QC, using a minimum of three batches.
Accuracy: The between-batch mean value should be within ±15% of the nominal value at
low, medium, and high QC concentrations and should not deviate by more than ±20% at
the LOQ.
Sensitivity: The lowest standard should be accepted as the limit of quantitation of the
method if the between-batch CV at the LOQ QC is #20%.
Specificity: The responses of interfering peaks at the retention time of the analyte should
be less than 20% of the response of an LOQ standard. Responses of interfering peaks at
the retention time of the internal standard should be #5% of the response of the
concentration of the internal standard to be used in studies.
Stability: Long-term, short-term, freeze and thaw, stock solution, and autosampler
stability data should meet the criteria specified in the SOP.
V. IN-STUDY VALIDATION
Assays of all samples of an analyte in a biological matrix should be completed within the time
period for which stability data are available. In general, analysis of biological samples can be done
with a single determination without duplicate or replicate analysis if the assay method has
acceptable variability as defined by validation data. This is true for procedures where precision
and accuracy variabilities routinely fall within acceptable tolerance limits. For a difficult
procedure with a labile analyte, where high precision and accuracy specifications may be difficult
to achieve, duplicate or even triplicate analyses may be recommended for better estimate of
analyte.
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