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.

中文翻译：

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新型LC-MS / MS方法分析人血浆中的二甲双胍和卡格列净：在药代动力学研究中的应用

开发了一种高灵敏度和选择性的液相色谱/串联质谱（LC-MS / MS）方法，并已验证该方法可用于同时批准最近批准的口服降血糖混合物；使用盐酸普萘洛尔（PPL）和他达拉非（TDF）作为内标（IS）分别测定人血浆中的二甲双胍（MET）和卡格列净（CFZ）。使用乙腈诱导的蛋白质沉淀提取分析物，然后使用乙酸乙酯进行液-液提取。反相HPLC使用C18分析柱（内径50 mm×4.6 mm，5 µm）进行，简单的等度流动相由0.1％甲酸和乙腈（60：40，v / v）组成。检测采用电喷雾电离技术在三重四极杆质谱仪上进行，并在多反应监测（MRM）中进行操作，在正离子模式下，MET，CFZ，PPL和TDF分别具有m / z 130.2→60.1，m / z 462.3→191.0，m / z 260.2→183.0和m / z 390.2→268.2的跃迁。在MET线性浓度范围为50-5000 ng / mL，CFZ为10-1000 ng / mL的线性浓度范围内，在5分钟内进行了分析。该方法已根据FDA的生物分析方法指南进行了验证。所提出的方法对所有被研究药物的回收率均高于90.0％，准确度为88.14–113.05％，相对标准偏差低于10.0％。取得的令人鼓舞的结果使已开发的LC-MS / MS方法可以成功地应用于目标药物对埃及健康志愿者的口服给药后的药代动力学研究。