Method Development and Validation of Anabasine and Nornicotine in Human Plasma by LC-MS/MS

Aim: For the Method development and validation of Anabasine Nornicotine in human plasma LC-MS/MS technique is used.
 Methodology: Operated positive electron spray ionization which removes hardness ions. Metoprolol used as an internal standard. Mass analyzers are to determine mass charge ratio. Liquid chromatography separates mixture components basis differences affinity for stationary mobile phase. It undesired impurities. increases reproducibility, sensitivity, robustness, detection low-level proteins. C18 Column (Phenomenex Luna C18, 5 μm, 50*4.6mm ID) high resolution peak area. Calibration curve constructed with help linear regression. During entire process no instability found.
 Results: m/z values 162.84(Q1), 134.68(Q3) 148.90(Q1), 79.82(Q3). Back calculated at different concentrations from 1-1000ng/ml their accuracy range 97-105% 93-108% Nornicotine. In precision by Intra-batch inter-batch. multi-step preparation, consider n=6 intra-batch n=18 inter-batch obtained between nominal i.e., 90-110%. each analyte coefficients variation below criteria <15%. average recovery rate 98.3% 94.3%.