Faculty of Pharmacy, Jawaharlal Nehru Technological University, Kakinada 533003, Andhra Pradesh, India; Vignan Pharmacy College, Vadlamudi 522213, Andhra Pradesh, India.
V. V. Institute of Pharmaceutical Sciences, Gudlavalleru 521356, Andhra Pradesh, India.
J Pharmacol Toxicol Methods. 2022 Jul-Aug;116:107186. doi: 10.1016/j.vascn.2022.107186. Epub 2022 May 26.
The combination of Ivabradine (IVA) and Metoprolol (MET) was approved by US-FDA for symptomatic treatment of chronic stable angina pectoris. Hence, a potential analytical method that can simultaneously quantify these two drugs is required. In view of this, a novel and fully validated LC-ESI-MS/MS method has been established for the quantification of IVA and MET in rat plasma. Analytes and their deuterated analogues were quantitatively extracted from rat plasma by protein precipitation technique. The analytes were separated using acetonitrile-water consisting 0.1% orthophosphoric acid buffer (30:70 v/v) as a mobile phase with a flow-rate of 1.0 mL/min and 5 min run time on Waters, X-Bridge-C18 (150× 4.6 mm, 3.5 μm) analytical column. The multiple reaction monitoring transitions, m/z 638.14 → 124.22 for IVA, 498.33 → 110.59 for MET; 644.37 → 130.41 for IVA-D6 and 504.46 → 116.28 for MET-D6 were chosen to achieve high selectivity in the analysis. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 0.1-1.5 ng/mL for IVA, 1.0-15.0 ng/mL for MET, with satisfactory precision and accuracy according to USFDA guidelines. Accuracy was within 99.71-100.3% and 99.9-100.31% for IVA and MET. The intra- and inter-day precision ranged between 0.048 and 12.68% and 0.1-2.66% CV for IVA and MET respectively. Further, the results of the pharmacokinetic parameters including C, t, AUC, AUC and t values of drugs indicated that the method is useful for successful quantification of the drugs in rat plasma. The developed method is significant and is useful for simultaneous quantification of IVA and MET.
伊伐布雷定（IVA）和酒石酸美托洛尔（MET）联合用药已获得美国食品药品监督管理局（US-FDA）批准，用于治疗慢性稳定性心绞痛。因此，需要建立一种能够同时定量检测这两种药物的分析方法。鉴于此，本文建立了一种新型、完全验证的 LC-ESI-MS/MS 方法，用于定量检测大鼠血浆中的 IVA 和 MET。采用蛋白沉淀技术从大鼠血浆中定量提取分析物及其氘代类似物。使用乙腈-水（含 0.1%正磷酸缓冲液，30:70 v/v）作为流动相，流速为 1.0 mL/min，在 Waters，X-Bridge-C18（150×4.6 mm，3.5 µm）分析柱上，运行时间为 5 min，实现了分析物的分离。多反应监测转换，m/z 638.14→124.22 用于 IVA，498.33→110.59 用于 MET；644.37→130.41 用于 IVA-D6，504.46→116.28 用于 MET-D6 被选为实现分析物高选择性的检测方法。与 USFDA 指南一致，该方法在 IVA 的浓度范围为 0.1-1.5 ng/mL、MET 的浓度范围为 1.0-15.0 ng/mL 时，灵敏度显著提高，线性良好，具有良好的精密度和准确性。IVA 和 MET 的准确度分别在 99.71-100.3%和 99.9-100.31%之间。IVA 和 MET 的日内和日间精密度分别在 0.048-12.68%和 0.1-2.66%CV 之间。此外，药物的药代动力学参数（C、t、AUC、AUC 和 t 值）结果表明，该方法可用于成功定量检测大鼠血浆中的药物。该方法意义重大，可用于 IVA 和 MET 的同时定量检测。
