Abstract

Establishing and validating a sensitive and accurate LC-MS method for quantifying Lenacapavir (LCV) in rat plasma using D6-LCV as the internal standard. This article presents an overview of the bioanalytical LC-MS method, utilizing a Waters Symmetry C18 column, 150 × 4.6 mm, 3.5 μm and an organic mobile phase comprising acetonitrile and 0.1% formic acid buffer in a ratio of 20:80. The calibration curve for LCV exhibited a linearity range of 5–100 ngml−1 (r2 = 0.9999). Liquid–liquid extraction was employed to recover LCV from rat plasma, resulting in recovery percentages of 98.97%, 99.51%, and 99.49% at three different concentration levels. LCV remained stable during storage under various conditions. Pharmacokinetic analysis yielded key parameters, including a half-life of 120 hours and a time to reach a maximum concentration of 4 hours. LCV and IS were identified using proton adducts in the LC-MS analysis at m/z 969.32/509.15 and 975.28/515.07, respectively, by employing positive mode multiple reaction monitoring. This comprehensive evaluation demonstrates that the method meets stringent criteria for system specificity, linearity, and accuracy, all well within the predefined acceptance limits. Its adaptability for the precise determination of LCV positions it as an invaluable tool in the field of bioanalysis, expanding its clinical utility.

Key words: Lenacapavir, pharmacokinetics, bio-analytical, rat plasma, LC-MS/MS.