Selective Reaction Monitoring of Negative Electrospray Ionization Acetate Adduct Ions for the Bioanalysis of Dapagliflozin in Clinical Studies

Shrivastav

Biomedical Chromatography

et al. 2018

A single LC–MS/MS assay has been developed and validated for the simultaneous determination of metformin and dapagliflozin in human plasma using ion‐pair solid‐phase extraction. Chromatographic separation of the analytes and their internal standards was carried out on a reversed‐phase ACE 5CN (150 × 4.6 mm, 5 μm) column using acetonitrile–15 mm ammonium acetate, pH 4.5 (70:30, v/v) as the mobile phase. To achieve higher sensitivity and selectivity for the analytes, mass spectrometric analysis was performed using a polarity switching approach. Ion transitions studied using multiple reaction monitoring mode were m/z 130.1 [M + H]+/60.1 for metformin and m/z 467.1 [M + CH3COO]−/329.1 for dapagliflozin in the positive and negative modes, respectively. The linear calibration range of the assay was established from 1.00 to 2000 ng/mL for metformin and from 0.10 to 200 ng/mL for dapagliflozin to achieve a better assessment of the pharmacokinetics of the drugs. The limit of detection and limit of quantitation for the analytes were 0.39 and 1.0 ng/mL for metformin and 0.03 and 0.1 ng/mL for dapagliflozin, respectively. There was no interference of plasma matrix obtained from different sources, including hemolyzed and lipemic plasma. The method was successfully applied to study the effect of food on the pharmacokinetics of metformin and dapagliflozin in healthy subjects.

Section: Resultsmentioning

confidence: 92%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Simultaneous quantitation of metformin and dapagliflozin in human plasma by LC–MS/MS: Application to a pharmacokinetic study

Shrivastav

Biomedical Chromatography

et al. 2018

“…S. Madhavi et al [31] has described a rapid, an accurate and precise Ultra Performance Liquid Chromatography (UPLC) method for simultaneous estimation of Saxagliptin and Dapagliflozin in its tablet dosage form (10mg Dapagliflozin and 5mg Saxagliptin). The UPLC method was developed using 2.1 × 100 mm, reverse phase C18 column (Acquity UPLC ethylene bridge hybrid (BEH) C18 1.7 μm).…”

Section: Rp-uplc Methods For Determination Of Dapagliflozinmentioning

confidence: 99%

A Review on Analysis of Dapagliflozin.

Harnaskar¹,

.²,

.³

et al. 2018

JCPR

Dapagliflozin is a sodium-glucose co transporter 2 inhibitor (SGLT2) as a new class of oral antidiabetic drugs. It is indicated for treatment of Diabetes mellitus type 2, either alone or in combination with other oral hypoglycaemic agents. The aim of this review is to focus on update of determination of Dapagliflozin in bulk and in pharmaceutical preparations using chromatographic and spectrophotometric methods. Dapagliflozin is estimated by RP-HPLC, UV, RP-UPLC, LC-MS methods. This review provides detailed information on separation conditions for Dapagliflozin alone, in the presence combination with other drugs and in presence of its degradation products. KEYWORDS Dapagliflozin, RP-HPLC, UV, RP-UPLC, LC-MS/MS. mobile phase with the flow rate of 1 ml/min (milliliter/minute). The effluent was detected at 222 nm (nanometer) using photo diode array detector. The retention time of Dapagliflozin API and Dapagliflozin tablet were 3.160 min (minute) and 3.067 min (minute) respectively. Linearity for Dapagliflozin was established in the range of 50-150µg/ml (microgram/milliliter) (R2 = 0.99) respectively. The % recoveries of Dapagliflozin API and tablet were found to be in the range of 99.00-99.99 % and 98.50-99.99 % respectively. Precision studies were carried out and the relative standard deviation values were less than two. LOD and LOQ were noted to be 5.14 µg/ml and 15.6 µg/ml for API. The method was found to be robust. The proposed method was found to be specific, accurate, precise and robust can be used for estimation of Dapagliflozin in API and Pharmaceutical dosage form. Subrata Sarkar et al. [5] developed and validated a new simple, rapid, efficient and reproducible reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of Dapagliflozin in Tablet dosage form. The separation was carried on a Symmetry C18, 25cmx 4.6mm i.d. 5μm, Particle size column. Mobile phase was mixture of Methanol: ACN: % OPA in a ratio of 75:25:05 v/v/v at a flow rate of 1.0ml/min. Dapagliflozin shows maximum absorbance at 246nm. Dapagliflozin was resolved at 2.797minutes. The Dapagliflozin drug was linear in the concentration range of 0-70μg/ml and correlation coefficient was 0.99. LOD & (LOQ) were found to be 0.04 & 0.12 μg/ml respectively. The developed method is also found to be simple, precise, accurate, specific, robust and rapid for the estimation of Dapagliflozin in bulk and tablet dosage form. Manasa Sangapati et al. [6] reported an accurate, precise, specific and rapid RP-HPLC method for the determination of Dapagliflozin in API. The method was developed and validated as described in ICH guidelines. Better separation of the drug was performed by BDS column (250×4.5mm, 5μ). Mixture of ortho phosphoric acid and acetonitrile (45:55 v/v) as a mobile phase at a flow rate of 1ml/min. Eluent was monitored using PDA detector at 245nm. The developed method was validated for different parameters such as linearity, accuracy, precision, limit of detection (LOD), limit of Quantitation (LOQ), robustness. The re...

“…There are some reported spectrophotometric methods for DGF assay either in raw material or in its pharmaceutical dosage forms [5][6][7][8][9] , however these methods have low sensitivity and cannot be applied for analysis of DGF in complexed biological samples such as human plasma. In the published literature, there are some reported high performance liquid chromatography (HPLC) methods for analysis of DGF in both its pure and tablet forms, as well as in biological fluids [10][11][12][13][14][15][16][17][18][19] , however the reported HPLC methods are complex and expensive. Therefore, a new simple and sensitive spectroscopic method was required to achieve the aim of this study.…”

Section: Introductionmentioning

confidence: 99%

New spectrofluorimetric analysis of dapagliflozin after derivatization with NBD‐Cl in human plasma using factorial design experiments

et al. 2019

A new validated spectrofluorimetric method was proposed for dapagliflozin (DGF) analysis in bulk, plexin its commercially available tablets and in spiked human plasma.The proposed spectrofluorimetric method depended on the formation of a fluorescent complex soluble in organic liquids by a substitution reaction between 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) reagent and DGF in aqueous buffered solution at pH 7. The fluorescence intensity was measured at 522 nm after excitation at 453 nm. The high selectivity of the proposed method allowed analysis of DGF in dosage form and human plasma samples with average recovery values of 99.84 ± 1.38% and 98.71 ± 1.80%, respectively, without any interference from matrix components. The calibration range was 50-1000 ng ml −1 . The limit of detection (LOD) and limit of quantitation (LOQ) were 14.24 ng ml −1 and 43.14 ng ml −1 , respectively. The estimated relative standard deviation values were lower than 2.0%, this showed the excellent precision at both levels. Factorial design was used to get the optimum method conditions for the analysis of the resulting DGF fluorescence complex in different matrices. The proposed method could be used in routine analysis of DGF in quality control laboratories. Also, it could be used to assay DGF in human plasma and be applied for pharmacokinetic investigation of DGF.