Anorethidrani disuccinate (ACP) is a domestically designed A-decarbonized steroid that is currently being investigated in Phase I clinical trials for the treatment of solid tumors. Only the parent drug exhibited antitumor activity; its sterol metabolite M2 showed obvious antiestrogenic effects. We have developed a rapid, sensitive, and robust liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the direct quantification of ACP and a chemical derivatization method that can be used to quantify M2 derivatized with glycidyl trimethyl ammonium chloride (GTMA). A simple protein precipitation procedure was performed to quantify ACP. Injections were obtained within 3.5 min on an Eclipse Plus Phenyl-Hexyl column (50 mm × 2.1 mm i.d., 1.8 μm) with gradient elution; the calibration curve was linear over the range of 2.00–8000 ng/mL. For quantification of M2 in plasma, analytes were extracted by protein precipitation and converted to their GTMA derivatives at 60°C for 2 h at pH 12; the analytes and coelutants were separated on a Luna C8(2) column (50 mm × 2.0 mm i.d., 5.0 μm). The precision (RSD) and accuracy (RE) of the intra- and interday determinations were within 10%. The derivatization procedure is a novel method for sterol determination by LC-MS/MS. The results confirmed the usefulness of this method for characterizing the pharmacokinetic profiles of ACP and its major metabolite M2 in a Phase I pharmacokinetic study.

琥珀酸二氨基蒽酯（ACP）是一种国内设计的A-脱碳类固醇，目前正在I期临床试验中用于治疗实体瘤。只有母体药物表现出抗肿瘤活性。其固醇代谢物M2具有明显的抗雌激素作用。我们已经开发了一种快速，灵敏且稳定的液相色谱-串联质谱（LC-MS / MS）方法，用于ACP的直接定量，以及一种化学衍生化方法，可用于量化由缩水甘油基三甲基氯化铵（GTMA）衍生的M2 ）。进行了简单的蛋白质沉淀程序以定量ACP。在Eclipse Plus苯基己基色谱柱（50 mm×2.1 mm内径，1.8μm）上3.5分钟内进行梯度洗脱，得到进样液；校准曲线在2.00–8000 ng / mL范围内呈线性。为了定量分析血浆中的M2，通过蛋白质沉淀法提取分析物，并在60°C，pH 12下将其转化为GTMA衍生物；在Luna C8（2）色谱柱（50 mm×2.0 mm内径，5.0μm）上分离分析物和共洗脱物。日内和日间测定的准确性（RSD）和准确性（RE）在10％以内。衍生化程序是通过LC-MS / MS测定固醇的一种新方法。结果证实了该方法在I期药代动力学研究中表征ACP及其主要代谢产物M2的药代动力学特征的有用性。日内和日间测定的准确性（RSD）和准确性（RE）在10％以内。衍生化程序是通过LC-MS / MS测定固醇的一种新方法。结果证实了该方法在I期药物代谢动力学研究中表征ACP及其主要代谢产物M2的药物代谢动力学特征的有用性。日内和日间测定的准确性（RSD）和准确性（RE）在10％以内。衍生化程序是通过LC-MS / MS测定固醇的一种新方法。结果证实了该方法在I期药代动力学研究中表征ACP及其主要代谢产物M2的药代动力学特征的有用性。