Hydrophilic Interaction Liquid Chromatography-Electrospray Ionization Mass Spectrometry for Therapeutic Drug Monitoring of Metformin and Rosuvastatin in Human Plasma

Abstract

In this work a hydrophilic interaction liquid chromatography/positive ion electrospray mass spectrometric assay (HILIC/ESI-MS) has been developed and fully validated for the quantitation of metformin and rosuvastatin in human plasma. Sample preparation involved the use of 100 μL of human plasma, following protein precipitation and filtration. Metformin, rosuvastatin and 4-[2-(propylamino) ethyl] indoline 2 one hydrochloride (internal standard) were separated by using an X-Bridge-HILIC BEH analytical column (150.0 × 2.1 mm i.d., particle size 3.5 μm) with isocratic elution. A mobile phase consisting of 12% (v/v) 15 mM ammonium formate water solution in acetonitrile was used for the separation and pumped at a flow rate of 0.25 mL min−1. The linear range of the assay was 100 to 5000 ng mL−1 and 2 to 100 ng mL−1 for metformin and rosuvastatin, respectively. The current HILIC-ESI/MS method allows for the accurate and precise quantitation of metformin and rosuvastatin in human plasma with a simple sample preparation and a short a chromatographic run time (less than 15 min). Plasma samples from eight patients were further analysed proving the capability of the proposed method to support a wide range of clinical studies.

Bioanalytical Method Using Ultra-High-Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry (UHPL-CHRMS) for the Detection of Metformin in Human Plasma

“…Improved performance was obtained in this study using HRMS combined with a HILIC analytical column. In some previous studies, the LLOQ concentration was greater than 5 ng/mL [ 6 , 9 , 10 , 11 , 16 ]. In the Fachi et al paper, when ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS) was used to develop an analysis method for quantifying various drugs including metformin in human plasma, the concentration of LLOQ was 250 ng/mL [ 16 ].…”

“…Moreover, to elute polar compounds such as metformin, the HILIC analytical column offers improved sensitivity because the percentage of organic solvents in the mobile phase is high [ 17 ]. In previous metformin quantitative analysis studies [ 6 , 8 , 11 , 19 , 20 ], an isocratic method was generally used, but in this paper, a gradient method was used to shorten the time of peak elution. Therefore, the time was shortened by 1.5 min compared with the 3.5 min in the Mistri et al study, which was the longest time of previous studies.…”

“…Among previous studies of quantitative analysis for metformin in human plasma, the LLOQ values were ranged from 1 ng/mL to 50 ng/mL [ 6 , 8 , 9 , 11 , 21 ]. Of these, there was a study that used LC-MS/MS with a LLOQ concentration of 1 ng/mL, and the S/N ratio was 20 [ 8 ].…”

“…Various bioanalytical methods for the determination of metformin in human-derived materials have been developed, including gas chromatography (GC) with mass spectrometry [ 5 ] and liquid chromatography (LC) with UV detection [ 6 , 7 ] or mass spectrometry [ 8 , 9 , 10 ]. GC methods require a complex derivatization procedure before mass spectrometry to compensate chromatographic issues as low volatility [ 5 ].…”

“…GC methods require a complex derivatization procedure before mass spectrometry to compensate chromatographic issues as low volatility [ 5 ]. The reported LC methods with UV detection have shown higher lower limit of quantification (LLOQ) concentration levels [ 6 , 7 ]. Additionally, LC-tandem mass spectrometry suffers from several disadvantages, such as requiring long chromatographic times [ 8 , 9 , 10 ], a lack of sensitivity [ 9 , 10 , 11 ], and the use of complicated sample preparation procedures, including liquid-liquid extraction [ 9 ].…”