TITLE:
Development and Validation of Extraction Method for the Determination of UC781 in Cervicovaginal Fluid
AUTHORS:
Naser L. Rezk, Zhiqing Qiao, Majed Jeriasy
KEYWORDS:
HIV-Prevention; Cervicovaginal Fluid; Extraction; LC-MS; Nonnucleoside Reverse Transcriptase Inhibitor
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.4 No.12,
December
3,
2013
ABSTRACT: Liquid chromatography plays the important and critical role
in the field of clinical pharmacology of evaluating drugs in biological matrices.
Studying antiretroviral drugs in the female genital tract has important implications for using drugs
as a vaginal microbicide for prevention of HIV-1 sexual transmission. Accurate measurement
of drug levels is extremely important in optimizing drug concentration
in gel formulation. Extracting drugs from small volumes of viscous, proteinaceous
substances like cervicovaginal fluid (CVF) is a practically challenging
process. The proposed method was designed to introduce procedure for sample collection
and drug extraction procedure for CVF matrix before the chromatographic separation.
Based on this extraction method, we validated a reverse phase high performance liquid
chromatography with electrospray ionization mass spectrometry assay in order to
quantify UC781 in female genital tract compartment. The LC-MS method was validated
based on a novel extraction technique which proved to be efficient in reducing analyte
degradation with an average extraction efficiency of 72%. This method is accurate,
demonstrating an average accuracy over three QC (n = 30) concentrations ranging
from 99.9% to 106.1%. Average precision within-day and between-day ranged from
3.1% to 10.2% and 5.1% to 6.4%, respectively. We demonstrated that the
analyte was able to maintain its stability under various conditions using this extraction
method. The sample preparation, extraction, and the powerful liquid chromatography
and mass spectrometry can readily be applied for accurate quantification of similar
drugs in CVF.