Biomedical Engineering Reference
In-Depth Information
Method Development, Validation,
and Sample Analysis for Regulated
Quantitative Bioanalysis Using LC-MS/MS
Min Meng and Patrick K. Bennett
Abstract
There are three primary stages of the regulated bioanalysis process using
Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): method devel-
opment, method validation, and sample analysis including incurred sample reanaly-
sis (ISR). Robust and rugged LC-MS/MS methods are essential in support of drug
discovery, toxicology studies, and clinical trials. The development of a robust bio-
analytical method requires careful consideration of many critical parameters, such
as accuracy and precision, linearity, matrix effect, sensitivity, selectivity, stability,
throughput, and ruggedness (or reproducibility). Because bioanalytical data is criti-
cal for determining the safety and efficacy of a new drug, a bioanalytical method
must be validated following the Food and Drug Administration's (FDA) guidance
for the industry, the recommendations from various white papers and Standard
Operating Procedures (SOPs). A complete regulated method validation in biologi-
cal matrix minimally requires three interday precision and accuracy runs, various
short- and long-term solution and matrix stability assessments, extraction recovery,
dilution capability and linearity, extract stability, reinjection reproducibility, selec-
tivity and specificity, assessment of matrix effects, interference from concomitant
medications and prodrug/metabolites, etc. The final evaluation of any high quality
bioanalytical method is not complete until it passes the ultimate test of regulated
sample analysis and incurred sample reanalysis (ISR) which are also conducted
following the similar rules as validation.
M. Meng, Ph.D. (
*
)
Tandem Labs , 1121 East 3900 South Salt Lake City , UT 84124 , USA
e-mail: MengM@LabCorp.com
P. K. Bennett
Thermo Fisher Scienti fi c , San Jose , CA , USA
e-mail: patrick.bennett@thermo fi sher.com
