Quantification of Blonanserin in Human Plasma Using Liquid Chromatography- Electrospray Ionization-Tandem Mass Spectrophotometry-Application to Pharmacokinetic Study

Abstract

Background: A novel LC-MS/MS method was developed for the estimation of blonanserin (BLN) in spiked human plasma. Methods: Liquid-liquid extraction (LLE) technique was adopted for the extraction of BLN from human plasma and chromatographic separation was performed on a waters symmetry shield, C18 (4.6mm id x 50 mm) analytical column using 7 Mm ammonium formate and acetonitrile (30:70) v/v as mobile phase. Positive ion mode was selected to obtain the product ion m/z 367.24 → 296.19 for BLN and m/z 326.8 → 269.07 for clozapine as internal standard (IS). Results: Calibration curve was linear over the range of 0.01 to 5 ng/ml. Developed method was satisfactory validated as per US-FDA guidelines for the bioanalytical study because it exhibits excellent intra and interday accuracy with % nominal 90 → 98.4 %, precision %CV ≤ 2% in all quality control levels, shows acceptable % extraction recovery (95.15 % → 97.04 %), demonstrated excellent matrix and analyte selectivity (% interference=0), matrix effect (matrix factor 0.981 at LQC and 1.02 at HQC level) and satisfactory stability study results in all types (% nominal 93.91 % → 99.58 %). Along with pharmacokinetic study rabbit plasma samples also analysed for one batch of accuracy, precision, matrix effect and rationalized the suitability of the developed method in other preclinical sample species Conclusion: Present method was successfully optimised, validated and applied favorable for the pharmacokinetic study of marketed formulation in rabbit blood samples in single oral human equivalent dose. The applicability of the developed method undoubtedly can further extend during preclinical and clinical trials. Key words: Blonanserin, LCMS/MS, Validation, Plasma, Pharmacokinetic.