Mohamed Dalia, Elshahed Mona S, Nasr Tamer, Aboutaleb Nageh, Zakaria Ola
1Analytical Chemistry Department, Faculty of Pharmacy, Helwan University, Ein Helwan, Cairo, 11795 Egypt.
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, October University for Modern Sciences and Arts, 6 October City, 11787 Egypt.
BMC Chem. 2019 Jul 9;13(1):82. doi: 10.1186/s13065-019-0597-4. eCollection 2019 Dec.
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, ). 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.
建立了一种高灵敏度和高选择性的液相色谱/串联质谱（LC-MS/MS）方法，并进行了验证，用于同时测定最近批准的口服降糖混合物；分别以盐酸普萘洛尔（PPL）和他达拉非（TDF）为内标（IS），测定人血浆中的二甲双胍（MET）和卡格列净（CFZ）。采用乙腈诱导的蛋白沉淀法提取分析物，然后用乙酸乙酯进行液-液萃取。使用C18分析柱（50 mm×4.6 mm内径，5 µm）进行反相HPLC，流动相为简单的等度洗脱，由0.1%甲酸和乙腈（60:40）组成。在采用电喷雾电离技术的三重四极杆质谱仪上进行检测，以多反应监测（MRM）模式运行，在正离子模式下，MET、CFZ、PPL和TDF的质荷比转换分别为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。分析在5分钟内完成，MET的线性浓度范围为50-5000 ng/mL，CFZ的线性浓度范围为10-1000 ng/mL。该方法按照FDA生物分析方法指南进行了验证。所提出的方法对所有研究药物的回收率均高于90.0%，准确度在88.14-113.05%范围内，相对标准偏差低于10.0%。所取得的良好结果使得所开发的LC-MS/MS方法成功应用于目标药物口服给药给埃及健康志愿者后的药代动力学研究。药代动力学研究在伦理委员会批准后完成。
