Bioanalytical Method Development and Validation for Letrozole Tablet by LC-MS

Abstract

The main objective of this work is to develop rapid, selective and sensitive LC-MS/MS method that have short and simple extraction procedures, consume small amounts of solvent and biological fluid for extraction and a short turn-around time. Need for the development of method and validation by HPLC Method for letrozole for better accuracy, precision and higher sensitivity and recovery rate from human plasma sample.  The drug candidate letrozole API purchased from Biodeal Pharmaceutical (Himachal Pradesh). HPLC Model Shimadzu 2030, Column: 4.6mm×25cm Fine-pack C8 Steel Column, Software: Cromalion 7.2, Analytical balance: Shimadzu, pH meter: Toshcon Industries and LC-MS: Sciex QTRAP 4500 used for the entire research work. Optimization of chromatography in HPLC Method for letrozole using Mobile Phase as Water: Acetonitrile: Methanol (50:30:20 v/v/v) any many trials has been taken for variation in mobile phase, flow rate 1 mL/min and other parameters till the optimized chromatogram was observed, Extraction Technique LLE completed and observed mean value 4502.00, SD value 7.19 and % RSD were found 0.16 for method precision. The Mean value 4499, and SD 3.49 and %RSD 0.08 for System Suitability found. Recovery at LOQ level 2.35 SD, 0.26%RSD, Recovery at 100% level 3.33 SD, 0.07%RSD and Recovery at 150% level 3.31 SD, 0.05%RSD was observed. For method development of tablet formulation 2.5 mg the mean value observed 4489, 9.19 SD and % RSD 0.20 observed. The Regression Statistics data LLOQ and LOD for regression analysis were found 1.369 ng/ml and 0.637 ng/ml respectively, so the sensitivity of developed methods was very high rather than other available methods.
Keywords: Letrozole, Bio-analytical Method Development, LLOQ, Method Validation