Simultaneous Determination of 12 Anti-Obesity Drugs in Human Plasma by a 96-Well Protein Precipitation Plate Coupled with High-Performance Liquid Chromatography–Mass Spectrometry

Abstract

A 96-well protein precipitation plate coupled with high-performance liquid chromatography–mass spectrometry (HPLC- MS/MS) was used to simultaneously determine the concentrations of 12 weight-loss drugs in human plasma. The 96-well protein precipitation plate was chosen for simultaneous pretreatment of large sample volumes, making the whole process more efficient and faster. The analytes were separated on an Agilent Poroshell 120 EC-C18 column with a mixture of acetonitrile and water containing 0.1% formic acid using a gradient elution within 20 min of running time. Quantification was performed in positive-ion multiple-reaction-monitoring mode and using an electrospray ionization source. Under the optimized conditions, good extraction efficiency for almost all targets was achieved with negligible matrix interference in the subsequent HPLC-MS/MS analysis. A good linearity was obtained in the range of 0.10–20.0 ng × mL - 1 for bupropion (BUP), fenfluramine (FEN), fluoxetine (FLU), bisacodyl (BISA), sibutramine (SIBU), and rimonabant (RIMO); and 0.50–20.0 ng⋅mL - 1 for methylephedrine (MER), amphetamine (AMP), bumetanide (BUM), lovastatin (LOVA), simvastatin (SIM), and fenofibrate (FENO) with a correlation coefficient above 0.995. The method was fully validated with an accuracy of 75.63%–108.21%, matrix effect of 80.41%–117.71% except for FENO (76.07% at low concentration levels), and precision of 0.32%–13.12%. This method could characteristically and sensitively determine the concentration of 12 weight-loss drugs in plasma, provide support for clinically monitoring the development of adverse reactions, and guide the rational and appropriate use of weight-loss drugs for obese people.

Note:
Funding Information: This work was supported by the Hebei Medical Applicable Technology Tracking Project (GZ2020042), and the Natural Science Foundation of Hebei Province (H2018206122) .

Declaration of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethics Approval Statement: The involving human participants were approved by the Ethics Committee of Hebei Medical University (Approval Number 2020067).

Keywords: anti-obesity drugs, 96-well protein precipitation plate；high performance liquid chromatography-mass spectrometry；plasma