A new HPLC–MS/MS method for the simultaneous quantification of SGLT2 inhibitors and metformin in plasma and its application to a pharmacokinetic study in healthy volunteers

Abstract

Monitoring the plasma concentrations of metformin and sodium‐glucose cotransporter‐2 inhibitors (canagliflozin, dapagliflozin and empagliflozin) is essential for pharmacokinetic and bioequivalence studies and therapeutic monitoring. The present work therefore aimed to develop and validate a high‐performance liquid chromatography coupled to tandem mass spectrometry (HPLC–MS/MS) method for the simultaneous quantification of these drugs in human plasma. The analyses were performed using an Agilent 1200 HPLC system coupled to an Applied Biosystems API 3200 triple quadrupole MS/MS with electrospray ionization in positive ion mode. After one‐step protein precipitation of plasma with acetonitrile containing 0.1% formic acid, chromatographic separation was achieved on an Xbridge C18 column, with a mobile phase consisting of a gradient of water and acetonitrile, both containing 1 mm ammonium formate and 0.1% formic acid. Quantification was performed in multiple reaction monitoring mode using m/z 130.1 → 71.1 for metformin, m/z 462.0 → 191.2 for canagliflozin, m/z 426.1 → 167.1 for dapagliflozin and m/z 468.0 → 354.9 for empagliflozin. The proposed method was validated and demonstrated to be adequate for the quantification of metformin, canagliflozin, dapagliflozin and empagliflozin for clinical monitoring, pharmacokinetics and bioequivalence studies.

Metformin: A Prospective Alternative for the Treatment of Chronic Pain

“…Pharmacokinetic-pharmacodynamic modeling has shown a correlation between the time course of metformin concentrations in the portal vein and gut wall and hypoglycemic effect, instead of drug concentrations in liver ( Stepensky et al., 2002 ; Sun et al., 2011 ). The time to reach maximal plasma concentrations in human beings after metformin administration (T max ) is 1.5 to 2.7 h ( Caillé et al., 1993 ; Sambol et al., 1996 ; Wei et al., 2009 ; Zhang et al., 2014 ; McCreight et al., 2018 ), which follows a multiphasic pattern ( Graham et al., 2011 ), giving a peak plasma concentration (C max ) of 1.1 to 2.5 µg/ml ( Wei et al., 2009 ; Zhang et al., 2014 ; Dias et al., 2019 ), and a steady-state concentration range of 0.3 to 1.5 µg/ml (see Table 2 ). Plasma protein binding is negligible, and the drug is not metabolized ( Scheen, 1996 ).…”

“…Renal excretion of this drug from the tubule cell to the lumen is mediated through multidrug and toxin extrusion protein 1 (MATE1, SLC47A1 ) and 2K (MATE2K, SLC47A2 ) expressed on the apical membrane of the renal proximal tubule cells ( Sato et al., 2008 ; Tsuda et al., 2009 ; Ito et al., 2012 ). The elimination half-life (t 1/2 ) is between 1.5 and 7 h (Tucker et al., 1981; Caillé et al., 1993 ), or longer if renal function is impaired ( Caillé et al., 1993 ; Dias et al., 2019 ). Metformin is eliminated by glomerular filtration and tubular secretion.…”

Review of Chromatographic Methods Coupled with Modern Detection Techniques Applied in the Therapeutic Drugs Monitoring (TDM)

“…Metformin and sodium–glucose cotransporter-2 inhibitors (canagliflozin, dapagliflozin, and empagliflozin) were simultaneously determined in human plasma by LC-MS/MS for pharmacokinetic and bioequivalence studies and TDM [ 150 ]. Before chromatographic analysis, samples were prepared by protein precipitation with acetonitrile containing 0.1% formic acid.…”

“…The gliflozin doses were 100, 10, and 10 mg for canagliflozin, dapagliflozin, and empagliflozin, respectively. Data are presented as a mean ± standard deviation and n = 6 for each group [ 150 ].…”

Quantitative determination of canagliflozin in human plasma samples using a validated HPTLC method and its application to a pharmacokinetic study in rats

“…Some high‐performance liquid chromatography (HPLC) methods have been applied for the analysis of CNZ in rat or human plasma (Deepan, Rao, & Dhanaraju, 2017; Dudhe & Kamble, 2016; Iqbal, Khalil, Alanazi, & Al‐Rashood, 2015; Mabrouk, Soliman, El‐Azigy, & Mansour, 2020). Various LC–mass spectrometry (MS)/MS methods have also been reported for the determination and evaluation of pharmacokinetic profile of CNZ in rat or human or rabbit plasma (Bhatt & Rajkamal, 2018; Deepan, Rao, & Dhanaraju, 2019; Dias et al, 2019; Dong et al, 2018; Iqbal, Ezzeldin, et al, 2015; Kobuchi, Yano, Ito, & Sakaeda, 2016; Ramisetti, Atmakuri, Venkatay, & Adireddy, 2019; Saibaba, Pilli, Bimireddy, & Pandiyan, 2018; Shah et al, 2019; Shah & Shrivastav, 2018; Udhayavani, Sastry, Rajan, Tejaswi, & Rajani, 2018). LC–MS/MS‐based quantitative analysis of drugs in biological fluids is the preferred technique owing to its advantages such as sensitivity, selectivity and speed of analysis (Deepan et al, 2019; Iqbal, Ezzeldin, et al, 2015; Shah et al, 2019).…”