Simultaneous Determination of Canagliflozin and Metformin in Human Plasma by LC–MS/MS Assay and its Application to a Human Pharmacokinetic Study

Abstract

Objective:The main objective of this work was to develop a simple, rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of Canagliflozin and Metformin. Methods: Deuterated compounds of respective drugs were used an internal standard. Sample extraction was carried out using a simple Protein Precipitation (PP) technique. A C 18 column with an isocratic mobile phase composed of 5mM ammonium acetate with 0.01% formic acid and methanol were used for chromatographic separation. Results: The method was validated in the linearity range of 10.00-6028.00 ng/mL for Canagliflozin and 10.00-3027.00 ng/mL for Metformin. The precision and accuracy results over five concentration levels in five different batches were well within the acceptance limits. A variety of stability tests were executed in plasma and in neat samples are comply with the FDA guidelines. Conclusion: The proposed assay method is simple, rapid and sensitive for the simultaneous determination of Canagliflozin and Metformin in human plasma. This method was successfully used to quantitate the in vivo plasma concentrations obtained from a pharmacokinetic study.

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

“…However, the reported HPLC technique with UV detection was less sensitive than the proposed HPTLC technique (Deepan et al, 2017). The sensitivity of eight LC–MS/MS and two UPLC–MS/MS techniques was also better than proposed HPTLC assay but most of these methods were not better in terms of cost, accuracy and precision (Bhatt & Rajkamal, 2018; Deepan et al, 2019; Dong et al, 2018; Iqbal, Ezzeldin, et al, 2015; Kobuchi et al, 2016; Ramisetti et al, 2019; Saibaba et al, 2018; Shah et al, 2019; Shah & Shrivastav, 2018; Udhayavani et al, 2018). Overall, the proposed HPTLC technique for the estimation of CNZ in plasma has been found more simple, time saving and cost effective with good accuracy and precision compared with reported techniques.…”

“…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).…”

Liquid chromatographic methods for the analysis of canagliflozin: concise overview and greener assessment

“…5 Similarly, one-step PP techniques have been used to extract CNG from urine and plasma samples using protein-precipitating agents, such as acetonitrile and methanol. 6,7 The recovery of CNG was found to be in the range of 81–84%. A good recovery of CNG and reproducible results were achieved with the developed sample preparation method.…”

Development and Validation of Reverse Phase High Performance Liquid Chromatography Method for the Estimation of Canagliflozin in Bulk and its Pharmaceutical Formulation

“…The RP-HPLC method developed was targeted for quantifying Canagliflozin and validation of the designed method was carried out in accordance to the ICH guidelines for the parameters namely linearity, precision, specificity, accuracy, robustness, detection limit and quantitative limit. 16…”

“…They allow weight-reducing and effective glycemic control combined with a low risk of hyperglycemia. [2][3][4][5][6][7] Canagliflozin is white crystalline powder, almost odorless, slightly soluble in water, freely soluble in acetonitrile, methanol and is chemically named as (2S, 3R, 4R, 5S, 6R)-2-(3-{[5-(4-fluorophenyl) thiophen-2-yl] methyl}-4-methylphenyl)-6-(hydroxyl methyl) oxane-3, 4, 5-triol having molecular formula C 24 H 25 FO 5 S. A detailed survey of literature revealed the determination of canagliflozin by UV spectroscopy, 6 HPTLC, 8 HPLC 9-11 and LC-MS/ MS [12][13][14][15][16] in simultaneous estimation of drugs and in biological fluids (i.e. human and rat plasma).…”