This study established and validated an LC–MS/MS method for the ultrasensitive determination of cetagliptin in human plasma. Sample pretreatment was achieved by liquid–liquid extraction with ethyl acetate, and chromatographic separation was performed on an XB‐C₁₈ analytical column (50 × 2.1 mm, 5 μm) with gradient elution (0.1% formic acid in acetonitrile and 0.1% formic acid) at a flow rate of 1.0 mL/min. For mass spectrometric detection, multiple reaction monitoring was used, and the ion transitions monitored were m/z 421.2–86.0 for cetagliptin and m/z 424.2–88.0 for cetagliptin‐d3. Method validation was performed according to the U.S. Food and Drug Administration Bioanalytical Method Validation Guidance, for which the calibration curve was linear in the range of 50.0–2000 pg/mL. All of the other results, such as selectivity, lower limit of quantitation, precision, accuracy, matrix effect, recovery, and stability, met the acceptance criteria. The validated method was successfully applied in a microdose clinical trial to systematically investigate the pharmacokinetic profile of cetagliptin in healthy subjects. Both rapid absorption and prolonged duration demonstrate the potential value of cetagliptin for diabetes treatment.

中文翻译：

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超灵敏LC-MS / MS方法的开发和验证，用于定量测定人血浆中的西格列汀及其在微剂量临床试验中的应用

这项研究建立并验证了一种LC-MS / MS方法，用于超灵敏测定人血浆中的西格列汀。样品通过乙酸乙酯的液-液萃取进行预处理，并在XB-C ₁₈分析柱（50×2.1 mm，5μm）上进行色谱分离，并进行梯度洗脱（乙腈中0.1％甲酸和0.1％甲酸） ）的流速为1.0 mL / min。对于质谱检测，使用了多反应监测，所监测的西立列汀和m / z的离子跃迁为m / z 421.2-86.0头孢列汀-d3为424.2–88.0。方法验证是根据美国食品药品监督管理局生物分析方法验证指南进行的，其校准曲线在50.0–2000 pg / mL范围内呈线性。所有其他结果，如选择性，定量下限，精密度，准确度，基质效应，回收率和稳定性，均符合可接受标准。验证的方法已成功用于微剂量临床试验，以系统研究头孢列汀在健康受试者中的药代动力学。快速吸收和持续时间均证明了西格列汀在糖尿病治疗中的潜在价值。