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

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.

LC-MS/MS Determination and Pharmacokinetics Study of Apremilast after Oral Administration in Rabbits

“…Molecular form is C 22 H 24 N 2 O 7 S with mass of 460.5. 2-4 A through literature survey reveals that various analytical and bioanalytical methods were published to describe the quantification of Apremilast in active pharmaceutical ingredient and pharmaceutical dosage forms to study the purity, degradation products by UV-Spectrophotometric method, 5-6 FTIR, 7 RP-HPLC, [8][9][10][11][12][13][14][15][16][17][18][19][20][21] Pharmacokinetic studies in beagle dog plasma, 22 human plasma and cerebro spinal fluid, [23][24][25][26][27] rat plasma [28][29][30] by LC-MS/MS. The published methods by LC-MS/MS was calibrated 0.1-100 ng/m.L of apremilast in biological samples.…”