A novel, selective and sensitive method is developed for simultaneous estimation of canagliflozin and metformin and successfully applied to fast and fed pharmacokinetic studies in healthy Indian volunteers. The current study reports the development, optimization, and validation of liquid chromatography-mass spectrometry (LC-MS/MS) method for simultaneous quantification of canagliflozin and metformin in human plasma using deuterated canagliflozin D4 and metformin D6 as an internal standard (IS). The solid-phase extraction technique was employed where strata X polymeric reverse phase (30 mg-1 cc) SPE cartridges were used for the extraction of analytes and IS from plasma. The ACE 5 C18 column (50 x 4.6 mm, 5µ) was used to chromatograph the prepared samples. The mobile phase consisted of methanol and 5 mM ammonium trifluoroacetate in water, pH 5 (50:50, v/v) at a flow rate of 0.8 mL/min.

Detection was performed by positive ion Turbo ion spray in Multiple reaction monitoring (MRM) mode, monitoring the transitions m/z 461.9→m/z 191.1 and m/z 461.9→m/z 267.2, for quantification of canagliflozin. The response of canagliflozin fragments m/z 461.9→m/z 191.1 and m/z 461.9→m/z 267.2 was combined. Also, for metformin transitions were monitored at m/z 130.0→m/z 71.1. Full validation of the method was performed according to the United States Food and Drugs Administration (USFDA) guidelines. Linearity was in the range of 24.95-2806.55 ng/mL for canagliflozin and 24.99-3400.72 ng/mL for metformin. The mean extraction recovery of canagliflozin, canagliflozin D4, metformin, and metformin D6 was 77.240, 84.663, 66.747, and 67.449, respectively across four QC levels. This rapid method with the run time of 2.80 min allows the analysis of more than 400 plasma samples per day.

开发了一种新颖，选择性和灵敏的方法，用于同时估算卡格列净和二甲双胍，并成功地应用于健康印度志愿者的快速和进食药代动力学研究。本研究报告了使用氘化的canagliflozin D4和metformin D6作为内标（IS）同时测定人血浆中canagliflozin和metformin的液相色谱-质谱（LC-MS / MS）方法的开发，优化和验证。采用了固相萃取技术，其中使用了X层聚合反相（30 mg-1 cc）SPE柱用于从血浆中萃取分析物和IS。使用ACE 5 C18色谱柱（50 x 4.6 mm，5µ）对制备的样品进行色谱分离。流动相由甲醇和5 mM的三氟乙酸铵水溶液（pH 5（50：

通过多反应监测（MRM）模式中的正离子涡轮离子喷雾进行检测，监测m / z 461.9→ m / z 191.1和m / z 461.9→ m / z 267.2的跃迁，以定量卡那列净。卡那列净片段m / z 461.9→ m / z 191.1和m / z 461.9→ m / z 267.2的响应合并在一起。同样，对于二甲双胍，监测的m / z转化率为130.0→ m / z71.1。根据美国食品和药物管理局（USFDA）指南对方法进行了完全验证。卡那列净的线性范围为24.95-2806.55 ng / mL，二甲双胍为24.99-3400.72 ng / mL。在四个QC水平上，canagliflozin，canagliflozin D4，metformin和metformin D6的平均提取回收率分别为77.240、84.663、66.747和67.449。这种运行时间为2.80分钟的快速方法每天可分析400多个血浆样品。