School of Public Health, Hebei Medical University, Shijiazhuang 050017, China.
Jinan Center for Disease Control and Prevention, Jinan 250000, China.
Molecules. 2022 Jun 25;27(13):4087. doi: 10.3390/molecules27134087.
A high-performance liquid chromatography-tandem mass spectrometry method was established for the simultaneous determination of mycophenolic acid, mycophenolate mofetil, tacrolimus, rapamycin, everolimus and pimecrolimus in human whole blood by optimizing the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) preparation method. Whole blood was extracted into ethyl acetate, salted out with anhydrous magnesium sulfate, and purified with ethylenediamine-N-propyl silane adsorbent. The supernatant was evaporated under nitrogen until dry and finally reconstituted in methanol. Chromatographic separation was performed on an Agilent Poroshell 120 EC-C18 column in methanol (mobile phase A)-water (optimized for 0.1% acetic acid and 10 mM ammonium acetate, mobile phase B) at a 0.3 mL·min−1 flow rate. Electrospray ionization and positive ion multiple reaction monitoring were used for detection. The time for of analysis was 13 min. The calibration curves range of tacrolimus, rapamycin, everolimus and pimecrolimus were in the range of 1−100 ng·mL−1, mycophenolate mofetil in the range of 0.1−10 ng·mL−1 and mycophenolic acid at 10−1000 ng·mL−1. All correlation coefficients were >0.993. The coefficients of variation (CV, %) for inter-day and intra-day precision were less than 10%, while the spiked recoveries were in the range of 92.1% to 116%. Our method was rapid, sensitive, specific, and reproducible for the simultaneous determination of six immunosuppressants in human whole blood. Importantly, our approach can be used to monitor drug concentrations in the blood to facilitate disease treatment.
建立了一种高效液相色谱-串联质谱法，用于通过优化 QuEChERS（快速、简单、廉价、有效、坚固、安全）制备方法，同时测定人全血中的麦考酚酸、霉酚酸酯、他克莫司、雷帕霉素、依维莫司和吡美莫司。全血用乙酸乙酯提取，用无水硫酸镁盐析，用乙二胺-N-丙基硅烷吸附剂纯化。上清液在氮气下蒸发至干，最后用甲醇重新溶解。在甲醇（流动相 A）-水（优化为 0.1% 乙酸和 10mM 乙酸铵，流动相 B）中，以 0.3mL·min−1的流速在 Agilent Poroshell 120 EC-C18 柱上进行色谱分离。电喷雾电离和正离子多反应监测用于检测。分析时间为 13min。他克莫司、雷帕霉素、依维莫司和吡美莫司的校准曲线范围为 1−100ng·mL−1，霉酚酸酯的校准曲线范围为 0.1−10ng·mL−1，麦考酚酸的校准曲线范围为 10−1000ng·mL−1。所有相关系数均大于 0.993。日内和日间精密度的变异系数（CV，%）小于 10%，而添加回收率在 92.1%至 116%之间。我们的方法快速、灵敏、特异、可重复，可用于同时测定人全血中的六种免疫抑制剂。重要的是，我们的方法可以用于监测血液中的药物浓度，以促进疾病的治疗。
