Medical College, Qinghai University, 251 Ningda Road, Xining, 810001, Qinghai, People's Republic of China.
Department of Hepatopancreatobiliary Surgery Affiliated Hospital of Qinghai University, Xining, 810001, Qinghai, People's Republic of China.
Sci Rep. 2022 Feb 28;12(1):3301. doi: 10.1038/s41598-022-07278-1.
The total alkaloids extracted from the seeds of Sophora moorcroftiana (TAs-SM) have the potential to treat alveolar echinococcosis, a disease included by the WHO in a list of 17 key neglected diseases world-wide. The aims of the current study were first to develop a supercritical fluid extraction (SFE) method for optimizing TAs-SM extraction, and second, to develop an optimized method for evaluating TAs-SM pharmacokinetics in vivo. The Box-Behnken response surface method was used to optimize the extraction process, and ultra-high liquid chromatography coupled with high resolution electrospray mass spectrometry (UPLC-HR-ESI-MS) was used to determine the pharmacokinetics of TAs-SM in SD rats. The results indicated the following optimal SFE extraction conditions: pressure = 31 MPa, temperature = 70 °C, time = 162.18 min. With these parameters, total alkaloids could be extracted from each gram of S. moorcroftiana, with the total content being 68.88 μg. The linear range of UPLC-HR-ESI-MS is 0.78-200.00 ng/ml, R > 0.99, and the sample recovery is 99-113%. The precision, accuracy, selectivity and stability of the method meet the requirements of US FDA guidelines. To our knowledge this study is the first to establish an SFE method for extracting TAs-SM and the first to employ UPLC-HR-ESI-MS for measuring TAs-SM in rats. These findings provide important contributions for using TAs-SM in further drug development and clinical applications.
从苦参种子中提取的总生物碱(TAs-SM)有可能治疗泡型包虫病,该病被世界卫生组织列入全球 17 种重点被忽视疾病之一。本研究的目的首先是开发一种超临界流体提取(SFE)方法来优化 TAs-SM 提取,其次是开发一种优化的体内 TAs-SM 药代动力学评估方法。采用 Box-Behnken 响应面法优化提取工艺,采用超高效液相色谱-高分辨电喷雾质谱联用(UPLC-HR-ESI-MS)测定 SD 大鼠 TAs-SM 的药代动力学。结果表明,最佳 SFE 提取条件为:压力=31 MPa,温度=70°C,时间=162.18 min。在这些参数下,每克苦参可提取总生物碱,总含量为 68.88μg。UPLC-HR-ESI-MS 的线性范围为 0.78-200.00ng/ml,R>0.99,样品回收率为 99-113%。该方法的精密度、准确度、选择性和稳定性均符合美国 FDA 指南的要求。据我们所知,本研究首次建立了 SFE 法提取 TAs-SM,首次采用 UPLC-HR-ESI-MS 法测定大鼠 TAs-SM。这些发现为进一步开发 TAs-SM 药物和临床应用提供了重要贡献。