Development and Validation of a Liquid Chromatography/Mass Spectrometry Method for the Simultaneous Quantitation of Rosuvastatin and Ezetimibe in Human Plasma

Abstract

A simple, rapid, and sensitive LC/electrospray ionization (ESI)-MS method was developed and validated for the simultaneous determination of rosuvastatin (ROS) and ezetimibe (EZE) in human plasma. Following liquid-liquid extraction, the analytes and an internal standard, atorvastatin (ATO), were separated using an isocratic mobile phase comprising 0.1% (v/v) formic acid-methanol (20 + 80, v/v) on an RP-C18 column. Detection was performed on a mass spectrometer by selected ion monitoring using their respective [M-H]- ions, m/z 480 for ROS, m/z 408 for EZE, and m/z 557 for ATO. For both analytes, the method was linear in the range of 0.1 to 10 nglmL. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 4 min made it possible to determine many plasma samples/day. The validated LC/ESI-MS method can be used to study pharmacokinetics, bioavailability, and bioequivalence of combined dosage forms of ROS and EZE.

Simultaneous quantitation of rosuvastatin and ezetimibe in human plasma by LC–MS/MS: Pharmacokinetic study of fixed‐dose formulation and separate tablets

“…During method development, several organic solvents, including diethyl ether, dichloromethane, MTBE and their combinations, were tested. Use of diethyl ether and dichloromethane solvent mixture, as reported previously (Varghese & Ravi, ) in different volume ratios, afforded poor recovery for EZE (57–65%) at 0.01 and 0.03 ng/mL concentration levels, whereas in all other solvent systems, the recovery of ROS found was inconsistent and low compared with EZE (88–94%). Combination of dichloromethane and MTBE improved the recovery of ROS by about 5–7% but there was considerable variation at lower CSs in the calibration range.…”

“…LLE has been the technique of choice for the extraction of ROS and free EZE from plasma samples (Kang et al, ; Kim et al, ; Min et al, ; Varghese & Ravi, ). During method development, several organic solvents, including diethyl ether, dichloromethane, MTBE and their combinations, were tested.…”

“…Some other methods describe estimation of EZE and its glucuronide metabolite (Bahrami et al, ; Guo, Wang, He, Qiu, & Jiang, ), and EZE with other drugs (Abdelbary & Nebsen, ; El‐Bagary, Elkady, El‐Sherif, & Kadry, ; Karanam, Katakam, Chandu, Hwisa, & Adiki, ; Munaga, Valluru, Bonga, Rao, & Sharma, ). Simultaneous determination of ROS and EZE in human plasma has been the subject of several reports (Ashfaq, Ahmad, Khan, & Mustafa, ; Kang et al, ; Kim, An, Kim, & Shin, ; Min, Park, Jung, Chang, & Kim, ; Varghese & Ravi, ). Among these, two methods present only the results for method development and validation in spiked plasma samples (Ashfaq et al, ; Varghese & Ravi, ), whereas the emphasis of the other methods was to investigate the pharmacokinetics of ROS and EZE in healthy subjects with limited information on method development (Kang et al, ; Kim et al, ; Min et al, ).…”

“…Similarly, EZE has been determined in different biological fluids like human plasma (Li, Liu, Jia, Li, & Yu, 2006;Oliveira et al, 2006;Ucakturk, Ozaltin, & Kaya, 2009) and human serum, urine and feces (Oswald, Scheuch, Cascorbi, & Siegmund, 2006) using gas chromatography-mass spectrometry (Ucakturk et al, 2009) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) (Li et al, 2006;Oliveira et al, 2006;Oswald et al, 2006) Munaga, Valluru, Bonga, Rao, & Sharma, 2016). Simultaneous determination of ROS and EZE in human plasma has been the subject of several reports (Ashfaq, Ahmad, Khan, & Mustafa, 2013;Kang et al, 2017;Kim, An, Kim, & Shin, 2017;Min, Park, Jung, Chang, & Kim, 2017;Varghese & Ravi, 2013). Among these, two methods present only the results for method development and validation in spiked plasma samples (Ashfaq et al, 2013;Varghese & Ravi, 2013), whereas the emphasis of the other methods was to investigate the pharmacokinetics of ROS and EZE in healthy subjects with limited information on method development (Kang et al, 2017;Kim et al, 2017;Min et al, 2017).…”

Optimization of UPLC Method for Simultaneous Determination of Rosuvastatin and Rosuvastatin Degradation Products

“…[25][26][27] Several methods have also been published for the determination of rosuvastatin in biological fluids using RP-HPLC/UV detection 28 or LC/MS. 29 The objective of this work was to develop a new analytical method that could simultaneously be used for the determination of rosuvastatin and rosuvastatin degradation products in rosuvastatin pharmaceutical dosage forms. The method has to be stability indicating 30 and capable to detect all changes in product quality during stability testing at various stability conditions according to regulatory requirements (accelerated, long-term or stress stability).…”