Development and optimization of a fast method for the determination of statins in human plasma using microextraction by packed sorbent (MEPS) followed by ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)

Abstract

Miniaturized sample preparation techniques are very attractive in bioanalysis since a reduced size of sample is required. Regarding this matter, an ultra high-performance liquid chromatographic-tandem mass spectrometry (UHPLC-MS/MS) microextraction by packed sorbent (MEPS) method was developed and validated using the micro technique in a sample preparation procedure for simultaneous analysis of three statins in human plasma. The method was developed using multivariate experimental design and validated based on ANVISA (Brazillian Health Regulatory Agency) recommendations. The analytes were extracted from human plasma samples using a lab-made MEPS micro device using C18 as the sorbent and then separated on an Acquity UPLC BEH C18 column with a gradient elution of acetonitrile/ammonium formate (5 mmol L⁻¹) buffer solution at pH 4.5. The detection was performed on a MS/MS system with negative and positive electrospray ionization and selected reaction monitoring mode. The limits of quantitation for all analytes were within 10–20 ng mL⁻¹ in plasma. The developed method showed appropriate accuracy, precision, linearity and robustness over the linear range studied and proved to be suitable for the analysis of pravastatin, fluvastatin and atorvastatin in human plasma.