Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China.
Molecules. 2014 Jan 17;19(1):1189-200. doi: 10.3390/molecules19011189.
A sensitive, reliable and specific LC-MS-MS method was developed and validated for the identification and quantitation of all-trans retinoic acid (ATRA) in human plasma. Acitretin was used as the internal standard (IS). After liquid-liquid extraction of 500 μL plasma with methyl tert-butyl ether (MTBE), ATRA and the IS were chromatographed on a HyPURITY C18 column (150 mm×2.1 mm, 5 μm) with the column temperature set at 40 °C. The mobile phase was consisted of 40% phase A (MTBE-methanol-acetic acid, 50:50:0.5, v/v) and 60% phase B (water-methanol-acetic acid, 50:50:0.5, v/v) with a flow rate of 0.3 mL/min. The API 4000 triple quadrupole mass spectrometer was operated in multiple reaction monitoring (MRM) mode via the positive electrospray ionization interface using the transition m/z 301.4→123.1 for ATRA and m/z 326.9→177.1 for IS, respectively. The calibration curve was linear over the range of 0.45-217.00 ng/mL (r≥0.999) with a lower limit of quantitation (LLOQ) of 0.45 ng/mL. The intra- and inter-day precisions values were below 8% relative standard deviation and the accuracy was from 98.98% to 106.19% in terms of relative error. The validated method was successfully applied in a bioequivalence study of ATRA in Chinese healthy volunteers.
建立并验证了一种灵敏、可靠且特异的 LC-MS-MS 法，用于鉴定和定量人血浆中的全反式维甲酸（ATRA）。阿维 A 作为内标（IS）。取 500μL 血浆，用甲基叔丁基醚（MTBE）进行液-液萃取后，ATRA 和 IS 在 HyPURITY C18 柱（150mm×2.1mm，5μm）上进行色谱分离，柱温设定为 40°C。流动相由 40%相 A（MTBE-甲醇-乙酸，50:50:0.5，v/v）和 60%相 B（水-甲醇-乙酸，50:50:0.5，v/v）组成，流速为 0.3mL/min。API 4000 三重四极杆质谱仪通过正电喷雾电离接口以多反应监测（MRM）模式操作，用于 ATRA 的转换 m/z 301.4→123.1 和 IS 的 m/z 326.9→177.1。校准曲线在 0.45-217.00ng/mL 范围内呈线性（r≥0.999），定量下限（LLOQ）为 0.45ng/mL。日内和日间精密度值低于 8%相对标准偏差，准确度以相对误差计为 98.98%-106.19%。该验证方法成功应用于中国健康志愿者中 ATRA 的生物等效性研究。
