Pharma / Biotech

Challenges in simultaneous extraction and chromatographic separation of metformin and three SGLT-2 inhibitors in human plasma using LC-MS/MS.

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Challenges in simultaneous extraction and chromatographic separation of metformin and three SGLT-2 inhibitors in human plasma using LC-MS/MS.

J Pharm Biomed Anal. 2019 Jul 24;175:112790

Authors: Shah PA, Shrivastav PS, Sharma V, Yadav MS

Abstract
Optimization of extraction and chromatographic conditions is essential for the simultaneous analysis of drugs having different physico-chemical properties, especially for in vivo applications. The present work describes concurrent estimation of metformin (MET) and three sodium-glucose co-transporter 2 (SGLT-2) inhibitors namely canagliflozin (CANA), dapagliflozin (DAPA) and empagliflozin (EMPA) in human plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample clean-up was optimized using ion-pair solid-phase extraction with sodium lauryl sulphate on Strata-X extraction cartridges. Consistent recoveries were obtained by critically optimizing parameters such as washing and elution solvents during extraction. The extraction recovery ranged from 79 to 88% for all the analytes. Chromatographic separation were accomplished on a Cyano (150 × 4.6 mm, 5 μm) column within 5.0 min using acetonitrile and 10 mM ammonium formate buffer (75:25, v/v) as the mobile phase. The resolution factors between MET-EMPA, MET-DAPA, MET-CANA, DAPA-EMPA and CANA-DAPA were 4.92, 5.85, 7.13, 1.01 and 1.44, respectively. Calibration curves were linear in the concentration range of 2.00-2000, 3.00-3000, 0.20-200 and 1.50-1500 ng/mL for MET, CANA, DAPA and EMPA, respectively. Mass spectrometric analysis was performed using a polarity switching approach to achieve high sensitivity in multiple reaction monitoring mode. The method was validated using current regulatory guidelines and applied to study the pharmacokinetics of fixed-dose formulations of MET and SGLT-2 inhibitors in healthy subjects.

PMID: 31362250 [PubMed – as supplied by publisher]

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