Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A One-step Sample Processing Method in Combination With Hplc-Ms/Ms for the Simultaneous Quantification of Atorvastatin, Ezetimibe and Three Metabolites Including O-Hydroxyl Atorvastatin, P-Hydroxyl Atorvastatin, and Ezetimibe-Glucuronide in Human Plasma

Version 1 : Received: 19 June 2023 / Approved: 21 June 2023 / Online: 21 June 2023 (12:06:30 CEST)

A peer-reviewed article of this Preprint also exists.

Le, T.N.N.; Chuong, N.N.; Nguyen, T.D. A One-Step Sample Processing Method in Combination with HPLC-MS/MS for the Simultaneous Quantification of Atorvastatin, Ezetimibe and Three Metabolites including o-Hydroxyl Atorvastatin, p-Hydroxyl Atorvastatin, and Ezetimibe-Glucuronide in Human Plasma. Separations 2023, 10, 409. Le, T.N.N.; Chuong, N.N.; Nguyen, T.D. A One-Step Sample Processing Method in Combination with HPLC-MS/MS for the Simultaneous Quantification of Atorvastatin, Ezetimibe and Three Metabolites including o-Hydroxyl Atorvastatin, p-Hydroxyl Atorvastatin, and Ezetimibe-Glucuronide in Human Plasma. Separations 2023, 10, 409.

Abstract

A simple and sensitive liquid chromatography-tandem mass spectrometry (LC– MS/MS) method has been developed for the simultaneous determination of atorvastatin (ATOR), ezetimibe (EZM) and their three metabolites including o-hydroxyl atorvastatin (o-OH ATOR), p-hydroxyl atorvastatin (p-OH ATOR), and ezetimibe-glucuronide (EZM-G) in human plasma using benzyl paraben (BP) as internal standard (IS). The analytes and IS were ionized using ESI positive ion mode (ATOR, o-OH ATOR, and p-OH ATOR), ESI negative ion mode (EZM, EZM-G, and BP), and operated in multiple reaction monitoring (MRM) mode. They were next extracted by salting-out assisted liquid–liquid extraction with acetonitrile, and then analyzed by liquid chromatography on a reversed-phase chromatographic column (50 mm × 4.6 mm; 3.5 µm), using a mixture of ac-etonitrile and an acetic acid solution (0.5%) as the mobile phase, showing high extraction effi-ciency (>70%), and a minimized matrix effect. The method was satisfactorily validated, and showed excellent linearity over wide concentration ranges of 0.06–15 ng/mL, 0.6–150 ng/mL, 0.4–100 ng/mL, 0.12–30 ng/mL, and 0.05–3 ng/mL for EZM, EZM-G, ATOR, o-OH ATOR, and p-OH ATOR, respectively.

Keywords

atorvastatin; ezetimibe; o-hydroxyl atorvastatin; p-hydroxyl atorvastatin; ezetimibe-glucuronide; LC-MS/MS; SALLE; human plasma

Subject

Medicine and Pharmacology, Pharmacology and Toxicology

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