安宫牛黄丸体外安全性评价方法的研究

研究安宫牛黄丸中可溶性汞及三种毒性汞价态的体外溶出检测方法，探讨安宫牛黄丸体外安全性评价方法。通过试验研究提取溶剂、净化方式、提取、净化时间等因素对溶出的影响，确定最佳的提取方案。使用电感耦合等离子体质谱法测定可溶性汞，高效液相色谱法-电感耦合等离子体质谱法联用测定三种汞价态。安宫牛黄丸中可溶性汞及三种毒性汞价态的最佳提取方法为：用人工肠液在37 ℃下超声处理2 h，提取液静置20 h后取上清液。方法重现性好，最大程度地模拟药物在体内的吸收环境。安宫牛黄丸提取液中可溶性汞含量较高，但不含汞的有毒价态：甲基汞、乙基汞和二价汞。该方法可推广应用于含朱砂中成药中可溶性汞及其价态的体外溶出和测定。以毒性大的三种汞价态：甲基汞、乙基汞和二价汞作为体外指标，可在一定程度上评价安宫牛黄丸的安全性。

In Vitro Safety Evaluation Study of Angong Niuhuang Wan

To study a vitro extraction method to determine soluble mercury and mercury species in Angong Niuhuang Wan and investigate a vitro safety evaluation method, the best extraction scheme was determined after a great deal of conditioning experiments focusing on how the solvent，purification，duration of extraction and purification etc impact on mercury extraction. Soluble mercury was determined by inductively coupled plasma mass spectrometry and mercury species were determined by high performance liquid chromatography/inductively coupled plasma mass spectrometry. Result show that the best extraction method of soluble mercury and its species was ultrasonic extraction in artificial intestinal juice at 37 ℃ for 2 hours, letting it stand for 20 hours and taking the supernatant. The method was reproducible，which could simulate the human body environment to maximum extent. Methyl mercury, ethyl mercury and Hg2+ were not found in the extracts of Angong Niuhuang, although the content of soluble mercury was high. It was concluded that the method could be applied to the vitro extraction and determination of soluble mercury and mercury species in Chinese traditional medicines containing cinnabar. Methyl mercury, ethyl mercury and Hg2+ could be taken as the targets of in vitro safety evaluation of Angong Niuhuang.