Wang Yueyuan, Diao Zhipeng, Qu Yuchen, Fan Kai, Feng Chen, Lv Bo, Yu Yunli, Geng Zhou
Department of Pharmacy, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, Jiangsu, China.
Sci Rep. 2025 Jul 29;15(1):27728. doi: 10.1038/s41598-025-13396-3.
Omadacycline, a third-generation tetracycline antibiotic, exhibits time-dependent pharmacokinetics. The ratio of the 24-hour area under the concentration-time curve to minimum inhibitory concentration (AUC/MIC) serves as the primary pharmacokinetic/pharmacodynamic (PK/PD) index for omadacycline, demonstrating strong correlation with therapeutic efficacy, particularly in critically ill patient populations. To facilitate routine therapeutic drug monitoring (TDM) in clinical practice, a high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS) method was developed for quantification of omadacycline in human plasma. An Agilent 1260 series liquid chromatograph and an API 4000 triple tandem quadrupole mass spectrometer were used for the determination of omadacycline and endocannabinoids. The separation was performed on a Phenomenex KINETEX XB-C18 column (2.6 μm, 3 × 50 mm) with 0.1% formic acid-water and pure acetonitrile as the mobile phases. The LC-MS/MS separation was carried out using a gradient elution procedure at a flow rate of 0.4 mL/ min, and the total run time was 5 min. An electrospray ionization (ESI) was selected, and the detection ions of omadacycline and the internal standard (fexofenadine-d6) were determined by mass spectrometry scanning with multiple reaction monitoring (MRM) in positive ion mode as follows: m/z 557.4→453.4 and m/z 508.4→472.8, respectively. The established linear range (20-2000 ng/mL) effectively covers the plasma concentration range encountered in > 98% of clinical samples, making it suitable for routine therapeutic drug monitoring. The method demonstrated acceptable selectivity, recovery, and matrix effect. The results of intra-day precision and inter-day precision show that the relative standard deviation (RSD) is less than 10%, while the relative error (RE) is within ± 10.00%. In this study, we developed and validated a simple, sensitive and accurate method to quantify the concentration of omadacycline in human plasma using LC-MS/MS. This offers essential technical support for the rational clinical application of omadacycline.
奥马环素是一种第三代四环素类抗生素，具有时间依赖性药代动力学特征。24小时浓度-时间曲线下面积与最低抑菌浓度之比（AUC/MIC）是奥马环素的主要药代动力学/药效学（PK/PD）指标，与治疗效果密切相关，在重症患者群体中尤为明显。为便于临床实践中的常规治疗药物监测（TDM），开发了一种高效液相色谱-串联质谱（HPLC-MS/MS）法用于定量测定人血浆中的奥马环素。使用安捷伦1260系列液相色谱仪和API 4000三重串联四极杆质谱仪测定奥马环素和内源性大麻素。在Phenomenex KINETEX XB-C18柱（2.6μm，3×50mm）上进行分离，以0.1%甲酸水和纯乙腈作为流动相。LC-MS/MS分离采用梯度洗脱程序，流速为0.4mL/min，总运行时间为5分钟。选择电喷雾电离（ESI），在正离子模式下通过多反应监测（MRM）质谱扫描确定奥马环素和内标（非索非那定-d6）的检测离子，分别为m/z 557.4→453.4和m/z 508.4→472.8。所建立的线性范围（20-2000ng/mL）有效覆盖了>98%临床样本中遇到的血浆浓度范围，适用于常规治疗药物监测。该方法具有可接受的选择性、回收率和基质效应。日内精密度和日间精密度结果表明，相对标准偏差（RSD）小于10%，相对误差（RE）在±10.00%以内。在本研究中，我们开发并验证了一种使用LC-MS/MS定量测定人血浆中奥马环素浓度的简单、灵敏且准确的方法。这为奥马环素的合理临床应用提供了必要的技术支持。
