Background:Verapamil is an excellent drug used for the medication of hypertensionand trandolapril. It is an angiotensin-converting-enzyme inhibitor. Hence, it is an interestingmethod to develop a novel and reliable MS/UPLC strategy for the simultaneous development ofverapamil and trandolapril.Objective:This research study aims to develop a new, rapid, and sensitive UPLC-MS/MS methodfor the simultaneous estimation of verapamil and trandolapril in rat plasma using D6-verapamil and D6-trandolapril.Method:Separation was carried on column Symmetry C18 column (150x4.6 mm, 3.5 μm) usingisocratic elution with a buffer containing 1mL of formic acid in 1L of water and the mixture oftwo components like Buffer and Acetonitrile in the ratio of 80:20 as mobile phase with1mL/min flow rate at ambient temperature.Results:Analysis was performed within 5 minutes over a good linear concentration range from2.4 ng/mL to 48 ng/mL (r2 = 0.9993 ± 0.018) for verapamil and 10pg/mL to 200pg/mL (r2=0.9993± 0.006) for trandolapril .The extraction recoveries and matrix effect of verapamil andtrandolapril were 98.45, 99.95, 98.12, 99.66% and 98.27, 99.89, 97.78, 99.23% respectively, at differentQC concentration levels. Precision and recovery study results were determined within theacceptable limit. An electrospray ionization source was used to study verapamil and Trandolapril atm/z 454.72→182.16, 430.25→201.48, and IS for m/z 460.18→ 324.39, 436.28 → 340.52, whichwere ion pairs of mass analysis. This method has successfully been applied to explore verapamil(1.2mg/kg) with its internal standard (D6-Verapamil), trandolapril (0.005 mg/kg) with its internalstandard (D6-Trandolapril) extracted from rat plasma using liquid-liquid extraction.Conclusion:This manuscript focuses on the consistent evaluation of the key bioanalytical validationparameters, and the following are discussed: accuracy, precision, sensitivity, selectivity,standard curve, limits of quantification, range, recovery, and stability. These validation parametersare described, together with illustrations of validation methodology applied in the case ofchromatographic methods used in bioanalysis.

中文翻译：

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同时测定大鼠血浆中维拉帕米和群多普利的 UPLC-MS/MS 方法的开发和验证：在药代动力学研究中的应用

背景：维拉帕米是治疗高血压和群多普利的优良药物。它是一种血管紧张素转换酶抑制剂。因此，开发一种新颖可靠的 MS/UPLC 策略同时开发维拉帕米和群多普利是一种有趣的方法。目的：本研究旨在开发一种新的、快速、灵敏的 UPLC-MS/MS 方法用于同时测定维拉帕米。使用 D6-维拉帕米和 D6-群多普利对大鼠血浆中的群多普利进行分离。方法：在柱 Symmetry C18 柱（150x4.6 mm，3.5 μm）上进行分离，使用含有 1mL 甲酸的 1L 水和混合物的缓冲液进行等度洗脱以缓冲液和乙腈两种成分的比例为 80:20 作为流动相，室温下流速为 1mL/min。 结果：维拉帕米的线性浓度范围为 2.4 ng/mL 至 48 ng/mL (r2 = 0.9993 ± 0.018)，群多普利的线性浓度范围为 10 pg/mL 至 200 pg/mL (r2=0.9993± 0.006)，可在 5 分钟内完成分析。在不同的QC浓度水平下，维拉帕米和群多普利的提取回收率和基质效应分别为98.45、99.95、98.12、99.66%和98.27、99.89、97.78、99.23%。精密度和回收率研究结果在可接受的限度内确定。使用电喷雾电离源研究维拉帕米和群多普利 atm/z 454.72→182.16、430.25→201.48，以及 m/z 460.18→324.39、436.28→340.52 的 IS，它们是质量分析的离子对。该方法已成功应用于探索维拉帕米（1.2mg/kg）及其内标物（D6-维拉帕米）群多普利（0. 005 mg/kg）及其内标物（D6-群多普利）采用液-液萃取法从大鼠血浆中提取。结论：本手稿侧重于关键生物分析验证参数的一致性评估，并讨论以下内容：准确度、精密度、灵敏度、选择性、标准曲线、定量限、范围、回收率和稳定性。描述了这些验证参数，以及在生物分析中使用的色谱方法的情况下应用的验证方法的说明。