Novel LC–MS/MS method for analysis of metformin and canagliflozin in human plasma: application to a pharmacokinetic study

Abstract

Highly sensitive and selective liquid chromatography/tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous estimation of the recently approved oral hypoglycemic mixture; metformin (MET) and canagliflozin (CFZ) in human plasma using propranolol HCl (PPL) and tadalafil (TDF) as internal standards (IS), respectively. Analytes were extracted using protein precipitation induced by acetonitrile then liquid–liquid extraction was performed using ethyl acetate. Reversed phase HPLC was carried out using C18 analytical column (50 mm × 4.6 mm i.d., 5 µm) with a simple isocratic mobile phase composed of 0.1% formic acid and acetonitrile (60:40, v/v ). Detection was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in multiple reaction monitoring (MRM), with the transitions of m/z 130.2 → 60.1, m/z 462.3 → 191.0, m/z 260.2 → 183.0 and m/z 390.2 → 268.2 for MET, CFZ, PPL and TDF, respectively, in the positive ion mode. The analysis was carried out within 5 min over a linear concentration range of 50–5000 ng/mL for MET and 10–1000 ng/mL for CFZ. The method was validated in accordance with the FDA guidelines for bioanalytical method. All obtained recoveries were higher than 90.0% while the accuracy was in the range of 88.14–113.05% and the relative standard deviation was below 10.0% for all investigated drugs by the proposed method. The achieved promising results has allowed for the successful application of the developed LC–MS/MS method to a pharmacokinetic study of the target drugs after their oral administration to Egyptian healthy volunteers. The pharmacokinetic study was accomplished after the agreement of the ethics committee. Electronic supplementary material The online version of this article (10.1186/s13065-019-0597-4) contains supplementary material, which is available to authorized users.

Evaluation of influence of telmisartan on the pharmacokinetics and tissue distribution of canagliflozin in rats and mice

“…Different LLE solvents, including ethyl acetate, dichloromethane, and methyl tert-butyl ether, were explored. Finally, methyl tert-butyl ether was chosen as the solvent as it provided higher extraction recovery and was only required in low volumes (20)(21)(22).…”

Stability indicating RP-UPLC method development and validation for the simultaneous determination of ertugliflozin and metformin in plasma

“…Stability protocol was processed for ERGZ and MTFN in human plasma for the estimation of stability of the drugs under different conditions like long term stability at -20 • C for 20 days, short term stability at 25 • C for 48 hrs and freeze (at -20 • C) and thaw (at room temperature) cycles for three times (Mohamed et al, 2019;Babua et al, 2019). By equating the original fresh concentrations with the stability samples, %RSD and %stability was calculated for ERGZ and MTFN.…”

“…Thorough literature on ERGZ and MTFN reveals that no single method was reported on the stabilityindicating bio-analytical method by RP-UPLC in a plasma sample. All the reported methods were on UV-Visible spectrophotometer, RP-HPLC (Lakshmana et al, 2019;Ashutosh et al, 2015) and LC-MS/MS techniques (Mohamed et al, 2019;Scherf-Clavel et al, 2019) either in single or in combination. Hence, there was the need to develop new stabilityindicating RP-UPLC technique for the quanti ication of ERGZ and MTFN in human plasma.…”

Development and Validation of a Simple and Sensitive LC-MS/MS Method for Quantification of Metformin in Dried Blood Spot Its Application as an Indicator for Medication Adherence

“…Until now, only a few LC-MS/MS methods for determining MET have been published in biological samples, such as plasma and serum. 6,[12][13][14][15] Dried blood spot (DBS) screening tests were used in the 1960s to diagnose metabolic disorders (phenylketonuria) in infants. Dried Blood Spot (DBS) has recently become a popular choice for bio sampling and for the therapeutic drug monitoring.…”

“…Recently, few sensitive methods were reported for individual estimation of MET using LC-MS/MS. 6,[12][13][14][15][25][26][27][28] Only a few studies published the analysis of MET from DBS utilizing LC-MS/MS. 1,4,[29][30][31][32] This sampling technique (DBS) is also acceptable for infants, which reduces the risk of HIV infection and other infectious pathogens to a minimum.…”