Development and Validation of a Rapid, Simple and Precise LC-MS/MS Method for the Quantification of Vincristine

A rapid, precise, and economical LC-MS/MS method was developed and validated for the quantification of vincristine. The method utilizes electrospray ionization in positive mode with multiple reaction monitoring (MRM) for a precursor-to-product ion transition of m/z ۸۲۵.۳ > ۸۰۷.۳. Effective chromatographic separation was achieved using a Thermo Hypurity C۱۸ column (۵۰ mm x ۴.۶ mm, ۵ µm). The mobile phase consisted of ۲ mM ammonium formate with ۰.۲% formic acid in acetonitrile at a ratio of ۳۰:۷۰ (v/v), pumped at a flow rate of ۰.۵ mL/min. The retention time (RT) for vincristine was ۱.۲۱ minutes and the total run time was ۲.۵ minutes. The method was validated according to ICH Q۲(R۱) guidelines, with the Limit of Detection (LOD) and Limit of Quantitation (LOQ) determined to be ۳ ppm and ۱۰ ppm, respectively, calculated using standard regression statistics. Linearity was established over a concentration range of ۱۰– ۱۵۰ ppm (y = ۶۹۸.۹۸۳ x − ۲۶۰۶.۲۱; R² = ۰.۹۹۵۷۳۴). Accuracy (mean % recovery) across LOQ, ۵۰%, ۱۰۰%, and ۱۵۰% levels was within ۹۱–۱۰۶%, and precision (relative standard deviation [%RSD]) ranged from ۱.۲% to ۳.۱% (intraday and inter-day), meeting the acceptance criteria for routine analysis. The system suitability %RSD for the peak area was ۲.۲۷%, and the RSD for RT was ۰.۴۵%. The method was successfully applied to quantify vincristine loaded on drug-eluting stents (DES) and to evaluate in vitro release kinetics (DES extraction and release sample testing included in validation). This method is suitable for routine quality control and formulation development of vincristine-containing DES.

کلیدواژه ها:

Vincristine ، LC-MS/MS ، Validation ، ICH Q۲(R۱) ، LOD ، LOQ