Abstract

DEVELOPMENT AND VALIDATION OF HIGH PERFORMANCE LIQUID CHROMATOGRAPHY MASS SPECTROMETRIC METHOD FOR ESTIMATION OF LERCANIDIPINE ENANTIOMERS FROM HUMAN PLASMA USING SOLID PHASE EXTRACTION

Malini Sharma* and N. B. Sridharamurthy

ABSTRACT

A novel, simple, specific, sensitive and reproducible liquid chromatography mass Spectrometric (LC/MS-MS) assay method has been developed and validated for quantifying R and S enantiomers of Lercanidipine in human plasma biological matrix. The LC/MS-MS method includes use of Deuterated isotopes labeled (R-Lercanidipine D3 and S-Lercanidipine D3) as an internal standard (IS). Samples were extracted using solid phase extraction methodology. The detection was performed using API 4000 (AB Sciex) and PC based data system with Analyst 1.4.2 software. Chromatographic separation was achieved on Lux 3μ Cellulose-3, 150×4.6 mm column using isocratic mobile phase composition with (0.2% Aqueous Ammonia Solution: Acetonitrile: 55:45 V/V) at a flow rate of 1.0 mL/min at 40ºC column oven temperature, without splitter over a total run time of 16 min. Method validation was performed as per USFDA guidelines and the results met the pre-defined acceptance criteria. The calibration curve was linear over a concentration range of 0.041 to 25 ng/mL for R and S Lercanidipine with correlation value r2 ≥ 0.995. Inter-batch quality control samples accuracy for R and S Lercanidipine ranged from 99.84% to 100.6 % with inter-batch precision values of 0.87% to 2.70% during the course of validation, demonstrating acceptable assay accuracy and precision. Further experiments like; Matrix Effect, Bench-Top, Freeze-Thaw, Dry extract, Wet extract, Effect of Haemolysis, anticoagulant, Lipemic plasma, etc were conducted to assess the suitability of proposed method for intended usage. Method was found to be suitable for quantitation of R and S Lercanidipine clinical study samples.

Keywords: Lercanidipine; LCMS-MS; Human plasma, Method validation.