摘要: |
越南槐(Sophora tonkinensis)为豆科(Leguminosae)灌木,广泛分布于中国西南地区。其根为中药“山豆根”,常被用于治疗胃、牙龈、咽喉、肺等部位的炎症。为了充分利用越南槐药用植物资源,该研究建立了pH区带精制逆流色谱法分离制备越南槐地上部位中生物碱的方法。以二氯甲烷-甲醇-水(5:3:2, V/V)为溶剂系统,上相添加20 mmol·L-1盐酸作为固定相,下相添加10 mmol·L-1三乙胺作为流动相对越南槐地上部位中的总生物碱进行分离制备。分离得到的化合物结构经高分辨质谱,核磁共振数据分析鉴定。同时用PNPG法对分离得到的化合物进行α-葡萄糖苷酶的抑制活性的测试。结果表明:从1.2 g总生物碱中一次分离制备得到183 mg苦参碱和404 mg氧化苦参碱,用高效液相检测其纯度分别为98.7%和98.2%。α-葡萄糖苷酶的抑制活性的测试结果显示苦参碱和氧化苦参碱对α-葡萄糖苷酶都具有较弱的抑制作用,其IC50值分别为(724.60±90.93)mg·L-1和(115.90±14.05 )mg·L-1。该研究结果表明,尽管寻找到一个合适的溶剂系统比较耗时,但pH区带精制逆流色谱法仍是一种简单且能有效分离越南槐地上部位中生物碱的方法。 |
关键词: pH区带精制逆流色谱, 山豆根, 苦参碱, 氧化苦参碱, α-葡萄糖苷酶 |
DOI:10.11931/guihaia.gxzw202101027 |
分类号:Q946 |
文章编号:1000-3142(2022)09-1459-07 |
Fund project:国家自然科学基金(32060108,82060764); 广西自然科学基金(2018GXNSFAA294033,2018GXNSFAA281078); 中国博士后专项基金(200626); 广西植物研究所基本业务费(桂植业18004); 广西八桂学者专项(第三批); 广西创新驱动发展专项基金(桂科AA18118015); 广西植物功能物质研究与利用重点实验室开放基金(ZRJJ2020-2, ZRJJ2018-19); 省部共建药用资源化学与药物分子工程国家重点实验室项目(CMEMR2019-B13); 广东省重点领域研发计划资助(2020B1111110003); 中央引导地方科技发展专项(桂科20111010) |
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Separation and preparation of alkaloids with inhibitory activity of α-glucosidase from Sophora tonkinensis by pH-zone-refining counter-current chromatography |
LIANG Senlin1, 2, HUANG Yonglin2, HE Ruijie2, WANG Yafeng2,
YANG Bingyuan2, LI Dianpeng2, SI Hongbin1*
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1. College of Animal Science and Technology, Guangxi University, Nanning 530005, China;2. Guangxi Key Laboratory of Plant
Functional Phytochemicals and Sustainable Utilization, Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region
and Chinese Academy of Sciences, Guilin 541006, Guangxi, China
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Abstract: |
Sophora tonkinensis, a shrub of the Leguminosae family, is widespread in Southwest China.The root of S. tonkinensis, “ShanDouGen” in Chinese, is often used to treat inflammation of stomach, gingiva, throat, lung, and other parts. In order to make full use of S. tonkinensis, a pH-zone-refining counter-current chromatography(PZRCCC)method is established to separate and prepare alkaloids from its aerial parts. CH2Cl2-CH3OH-H2O(5:3:2, V/V)was selected as the solvent system of PZRCCC, 20 mmol·L-1 HCl was added to the upper aqueous stationary phase as a retainer and 10 mmol·L-1 triethylamine(TEA)to the organic mobile phase as an eluter. The structures of the isolates were elucidated by HRMS and NMR data. The α-glucosidase inhibitory activity of the isolates was determined by PNPG method. A total of 183 mg matrine and 404 mg oxymatrine were obtained from 1.2 g total alkaloids. The purity of matrine and oxymatrine were 98.7% and 98.2%, respectively, as determined by HPLC. Both matrine and oxymatrine had weak inhibitory effects on α-glucosidase with IC50 values of(724.60±90.93)mg·L-1and(115.90±14.05)mg·L-1, respectively. The present study demonstrated PZRCCC is a simple and efficient method for the separation of alkaloids from the aerial part of S. tonkinensis, although finding a suitable solvent system is a time-consuming procedure. |
Key words: pH-zone-refining counter-current chromatography, Sophora tonkinensis, matrine, oxymatrine, α-glucosidase |