A Selective and Sensitive Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) Method for Quantitative Bioanalysis of Efavirenz in Human Dried Blood Spots

摘要

The LC-MS/MS, tandem mass spectrometry, method described here allowed accurate and reproducible method for determination of efavirenz in human dried blood spots utilizing 30 μL of whole blood and short analytical run time of 4.0 min with an LLOQ of 1.0 ng/mL. The method was linear, accurate, precise, selective, rugged and met all of FDA suggested precision requirements for bio-analytical LC-MS/MS method validation guidelines. DBS assay can be developed to meet low sensitivity requirements in the low ng/mL quantitation range, with an advantage of low sample volume (30μL), reduced number of study animals, sample collection ease, reduced sample shipping and storage costs, and analyte matrix stability. Patients or animals blood with either low or high hematocrit levels can cause inconsistent spotting on cards. New DBS cards have been introduced using a fiberglass format that may overcome this issue. Coated sample collection cards (DMPK-B Card) have the potential to aid analyte stability and virus deactivation1. Automated liquid handling Systems (Tomtec Quadra 4.0) can be used to generate DBS during method validation to improve the efficiency of bio-analytical process. Theoretical plasma concentrations can be calculated from dried blood spot concentrations using formula: Plasma Concentration=[dried blood spot concentration/(1-haematocrit)]x fraction bound to plasma proteins.