Background: A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method had been developed for the quantification of acotiamide in rat plasma and been applied to pharmacokinetic studies. However, there was no LC-MS/MS method been developed for the determination of acotiamide in human plasma and its pharmacokinetic study.

Objective: A simple and fast LC-MS/MS method was established and validated for the quantification of acotiamide in human Received: plasma and was applied to a pharmacokinetic study.

Methods: Sample preparation was accomplished Revised: Accepted: through protein precipitation, and chromatographic separation was achieved on a Welch, Ultimate XB-C18 column (2.1×50 mm, 3 μm) with a security guard cartridge C18 using a binary gradient with DOI: mobile phase A (Methanol) and B (the solution of 10 mM Ammonium acetate with 0.1% Formic acid) at a flow rate of 400 μL/min.

Results: The retention time of acotiamide and its internal standard, acotiamide-d6 was 1.78 min and 1.79 min, respectively. The total run time was 4.0 min. The method was developed and validated over the concentration range of 0.500-100 ng/mL for acotiamide, with correlation coefficient greater than 0.9987. The extraction recovery was more than 108.43% and the matrix effect was not significant. The inter- and intra-day precisions were below 5.80% and accuracies ranged from 92.7 to 103.0%. Acotiamide was demonstrated to be stable in human plasma under the tested conditions.

Conclusion: The validated LC-MS/MS method was successfully applied to study the pharmacokinetic profiles of acotiamide in human plasma after oral administration and has achieved satisfactory results.

背景：已开发出一种液相色谱-串联质谱 (LC-MS/MS) 方法来定量大鼠血浆中的阿考替胺，并应用于药代动力学研究。然而，没有开发出用于测定人血浆中阿考替胺及其药代动力学研究的 LC-MS/MS 方法。

目的：建立并验证了一种简单快速的 LC-MS/MS 方法，用于定量人体接受：血浆中的阿考替胺，并将其应用于药代动力学研究。

方法：完成样品制备 修订：接受：通过蛋白质沉淀，并在 Welch Ultimate XB-C18 色谱柱（2.1×50 mm，3 μm）和安全保护柱 C18 上实现色谱分离，使用二元梯度和 DOI：流动相 A（甲醇）和 B（10 mM 醋酸铵与 0.1% 甲酸的溶液），流速为 400 μL/min。

结果：acotiamide 及其内标acotiamide-d6 的保留时间分别为1.78 min 和1.79 min。总运行时间为 4.0 分钟。该方法在 0.500-100 ng/mL 的阿考替胺浓度范围内开发和验证，相关系数大于 0.9987。提取回收率超过108.43%，基质效应不显着。日间和日内精密度低于 5.80%，准确度在 92.7% 至 103.0% 之间。在测试条件下，Acotiamide 被证明在人血浆中是稳定的。

结论：经验证的LC-MS/MS方法成功应用于阿考替胺口服给药后人血浆中的药代动力学特征研究，取得了满意的结果。