Determination of Aprepitant in Human Plasma by Using LC-MS/MS with Electrospray Ionization

Ravi Prakash Pvdls

doi:10.4172/jbb.1000143

Abstract

A precise, sensitive and high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of Aprepitant (APT) in human plasma was developed and validated using Quetiapine (QTP) as internal standard. The analyte and internal standard were extracted from human plasma using liquid-liquid extraction. Chromatographic separation was performed on Discovery C18 10 cm×4.6 mm, 5 μm column with an isocratic mobile phase composed of 5 mM Ammonium Acetate (pH 4.00):Acetonitrile (10:90), at a flow-rate of 0.9 ml/ min. The MS-MS detection was performed on a AB Sciex API 3200 tandem mass spectrometer operated in Multiple reaction monitoring (MRM) at positive mode at m/z 535.10/277.10 and 384.00/253.10 for APT and QTP respectively. A linear dynamic range of 10.004-5001.952 ng/ml for APT was evaluated with mean correlation coefficient (r) of 0.9991. The precision of the assay (expressed as coefficient of variation, CV) was less than 15% at concentrations of LQC, MQC, HQC and was less than 20% for LLOQQC. Percent recoveries for APT at high, middle and low quality control samples was found to be 71.9%, 68.0%, and 63.8% respectively and for internal standard 77.7%. The analyte was found to be stable throughout five freeze-thawing cycles, bench top, wet extract, dry extract, auto sampler and interim stability studies. Therefore, the proposed method was found to be suitable for the routine quality control analysis of Aprepitant in human plasma in bioequivalence studies.

Highlights

Aprepitant (APT) is chemically (5-[[2(R)-[1(R)-(3,5bistrifluoromethylphenyl)ethoxy]-3(S)-(4-fluorophenyl) morpholin4-yl]methyl]-2,4-dihydro-[1,2,4]triazol-3-one) and belongs to class of substance P antagonists (SPA)
neurokinin 1 (NK1) receptor antagonists might reverse the impairment of NK cell function found in HIV infection via antagonism against SP, whose effects are mediated through NK1 receptor [4]
The present study describes development and validation of a simple, specific, rapid and sensitive liquid chromatography - tandem mass spectrometry (LC-MS/MS) method for the determination of Aprepitant in human plasma with a limit of quantification (LOQ) of 10.004 ng/ml during a 2.5 min run time using QTP as internal standard

Summary

Introduction

Aprepitant (APT) is chemically (5-[[2(R)-[1(R)-(3,5bistrifluoromethylphenyl)ethoxy]-3(S)-(4-fluorophenyl) morpholin4-yl]methyl]-2,4-dihydro-[1,2,4]triazol-3-one) and belongs to class of substance P antagonists (SPA). It acts as an anti-emetic by blocking the neurokinin 1 (NK1) receptor. APT is used for the prevention of acute, delayed chemotherapy-induced and postoperative nausea and vomiting. It has little or no affinity towards 5-HT3 receptors but it is shown to increase the activity of 5-HT3 receptor antagonists such as ondansetron and the corticosteroid dexamethasone, which are used to prevent nausea and vomiting caused by chemotherapy [2]. APT is under evaluation as a new therapy in Neuro AIDS patients from the Integrated Preclinical and Clinical Program (IPCP) grant mechanism supported by the NIH at the Children’s Hospital of Philadelphia and University of Pennsylvania [5]

Methods

Results

Conclusion