Chen Lei, Zhao Linlin, Cai Wei, Shao Jing, Huang Xiaobin, Liu Yi
Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State Administration of TCM, China; Engineering and Technology Research Center for Chinese Materia Medical Quality of the Universities of Guangdong Province, Guangdong Pharmaceutical University, Guangzhou, China.
Department of Pharmacy, Hunan University of Medicine, Huaihua, China.
Biomed Chromatogr. 2019 Mar;33(3):e4447. doi: 10.1002/bmc.4447. Epub 2018 Dec 21.
In previous studies, it was revealed that ethyl acetate (EtOAc) extracts from Sophora flavescens Ait. improved glucose tolerance, reduced hyperglycemia, and restored insulin levels in diabetic patients. The aim of this study was to develop an accurate and sensitive UHPLC-MS method for simultaneous determination of flavonoids in EtOAc extracts of Kushen in rat plasma. Ethyl acetate-acetonitrile (2:1) was selected as the solvent to extract the four flavonoids from rat plasma. A BEH C column (2.1 mm × 100 mm, 1.7 μm) with a C guard cartridge was chosen as the separation plant using a gradient elution with acetonitrile (solvent A) and 0.1% formic acid (solvent B) in water. For all four analytes, the method showed good linearity (r > 0.991) in 1-500 ng/mL. The inter- and intra-day accuracy ranged from -13.78 to 7.19%, and the precision (RSD) was <8.75%. Recoveries of all four flavonoids ranged from 85.9 to 101.3%. According to the results of multitarget pharmacokinetic studies, four active flavonoids in EtOAc extracts from Kushen have similar absorption kinetics but very different metabolic kinetics, and a double peak phenomenon was observed in the concentration-time curve of norkurarinone, which is different from the previous study. In conclusion, detection and multitarget pharmacokinetic studies successfully determined active flavonoids after oral administration of EtOAc extracts from Kushen by an efficient, sensitive and selective UHPLC-MS method, and the results may provide a foundation for future studies of Kushen.
在先前的研究中，已揭示苦参乙酸乙酯（EtOAc）提取物可改善糖尿病患者的葡萄糖耐量、降低高血糖并恢复胰岛素水平。本研究的目的是开发一种准确、灵敏的超高效液相色谱-质谱联用（UHPLC-MS）方法，用于同时测定大鼠血浆中苦参乙酸乙酯提取物中的黄酮类化合物。选择乙酸乙酯-乙腈（2:1）作为从大鼠血浆中提取四种黄酮类化合物的溶剂。选用带有C保护柱套的BEH C柱（2.1 mm×100 mm，1.7 μm）作为分离柱，采用乙腈（溶剂A）和0.1%甲酸（溶剂B）的水溶液进行梯度洗脱。对于所有四种分析物，该方法在1-500 ng/mL范围内显示出良好的线性（r＞0.991）。日内和日间准确度范围为-13.78%至7.19%，精密度（相对标准偏差，RSD）＜8.75%。四种黄酮类化合物的回收率范围为85.9%至101.3%。根据多靶点药代动力学研究结果，苦参乙酸乙酯提取物中的四种活性黄酮类化合物具有相似的吸收动力学，但代谢动力学差异很大，去甲苦参酮的浓度-时间曲线中观察到双峰现象，这与先前的研究不同。总之，通过高效、灵敏和选择性的UHPLC-MS方法，检测和多靶点药代动力学研究成功测定了苦参乙酸乙酯提取物口服给药后的活性黄酮类化合物，研究结果可为苦参的后续研究提供基础。
