Patel Dhrumi, Vora Amisha, Wairkar Sarika, Yc Mayur
Shobhaben Pratapbhai Patel School of Pharmacy & Technology Management, SVKMs NMIMS, V.L. Mehta Road, Vile Parle (W), Mumbai, India, 400056, India.
J Mass Spectrom. 2021 Oct 16;56(11):e4789. doi: 10.1002/jms.4789.
This research aims to develop and validate a bioanalytical method for simultaneous estimation of an antidiabetic combination using LC-MS/MS in rat plasma. Nateglinide and metformin hydrochloride are commonly used combination for clinical management of Type 2 diabetes. Hence, simultaneous determination in plasma is essential for the rapid analysis of samples from the pharmacokinetic studies. Statistical optimization was carried out for liquid chromatography (LC) parameters and mass spectroscopic (MS) parameters by design of experiment (DoE) (Design Expert Version 11, Stat Ease Inc., USA) approach. A 3 full factorial design was used for optimization of LC parameters; %methanol, %formic acid, and flow rate were selected as independent variables, whereas peak area and tailing factor were considered as dependent variables for both drugs. Box-Behnken design was used to optimize MS parameters including drying gas flow rate, nebulizing gas flow rate, DL temperature, heat block temperature, and positive voltage as independent factors, and responses selected were [M + H] intensity of nateglinide and metformin hydrochloride. The [M + H] intensity of the optimized method for nateglinide and metformin hydrochloride were 2,462,838 and 11,873,826, respectively. The model was found significant for optimizing LC and MS parameters with p < 0.05 for both nateglinide and metformin hydrochloride. The optimized method was validated as per the ICH-M10 guideline, which was accurate, precise, and selective. The method was cost-effective and capable of quantitating concentrations in picogram levels for nateglinide and metformin hydrochloride simultaneously.
本研究旨在开发并验证一种生物分析方法，用于在大鼠血浆中同时测定一种抗糖尿病复方制剂，采用液相色谱-串联质谱法（LC-MS/MS）。那格列奈和盐酸二甲双胍是临床上用于2型糖尿病治疗的常用复方制剂。因此，血浆中的同时测定对于药代动力学研究样本的快速分析至关重要。通过实验设计（DoE）（Design Expert Version 11，Stat Ease Inc.，美国）方法对液相色谱（LC）参数和质谱（MS）参数进行了统计优化。采用三因素全因子设计优化LC参数；选择甲醇百分比、甲酸百分比和流速作为自变量，而将峰面积和拖尾因子作为两种药物的因变量。采用Box-Behnken设计优化MS参数，包括干燥气流量、雾化气流量、脱溶剂温度、加热块温度和正电压作为独立因素，选择的响应指标是那格列奈和盐酸二甲双胍的[M + H]强度。那格列奈和盐酸二甲双胍优化方法的[M + H]强度分别为2,462,838和11,873,826。发现该模型对于优化LC和MS参数具有显著性，那格列奈和盐酸二甲双胍的p均<0.05。按照ICH-M10指南对优化后的方法进行了验证，该方法准确、精密且具有选择性。该方法具有成本效益，能够同时定量测定那格列奈和盐酸二甲双胍皮克水平的浓度。
