摘要
建立了快速分离及鉴定葛花中活性化合物的方法。采用70%乙醇加热回流提取得到葛花提取物,以α-葡萄糖苷酶和乳酸脱氢酶作为生物靶分子,以超滤质谱技术筛选酶抑制剂。运用高速逆流色谱分离纯化所得酶抑制剂,以乙酸乙酯-乙醇-水(4.0∶0.5∶3.0,V/V/V)组成三元溶剂体系,从200 mg葛花粗提物中,一次性分离制备得到2.36 mg葛根素、8.57 mg鸢尾苷、5.34 mg染料木苷,经高效液相色谱分析其纯度均达到90.0%以上。该三个化合物均可与α-葡萄糖苷酶及乳酸脱氢酶亲和,具有潜在的抗糖尿病及抗脑卒中的活性。通过液-质联用技术及核磁共振波谱技术确定了每个化合物的结构。该方法简单、快速、高效,分离样品纯度高,适用于葛花中异黄酮类活性化合物的分离纯化及鉴定研究。
Using high speed counter-current chromatography(HSCCC) and ultrafiltration mass spectrometry to screen and separate α-glucosidase and lactate dehydrogenase(LDH) inhibitors from Puerariae Flos(P.Flos),a rapid method for the separation and preparation of active compounds in P.Flos was established.The extraction of P.Flos was obtained by using 60% ethanol heating reflux,then the purification was separated by HSCCC.A binary solvent system was composed of ethyl acetate-ethanol-water(4.0∶0.5∶3.0,V/V/V),and was used for separation and purification of the isoflavones in P.Flos.The enzyme inhibitors were screened using α-glucosidase and LDH as biomaterials with ultrafiltration spectrum.2.36 mg puerarin,8.57 mg tectorigenin and 5.34 mg genistin were obtained from 200 mg P.Flos crude extract.The purity of high performance liquid chromatography(HPLC) was above 90.0%.The three compounds can be associated with α-glucosidase and LDH,have potential antidiabetic and antiapoplexy activity.The structure of each compound was determined by 1H-NMR,13C-NMR and ultra-high-performance liquid chromatography-high-resolution mass spectroscopy(UPLC-MS/MS).This method is simple,fast and efficient,and the purity of the sample is high.It is suitable for the separation and purification of isoflavones in P.Flos,and provides a theoretical basis and technical platform for the development of natural medicines.
作者
吴桐
黄彧
侯万超
夏建丽
刘春明
李赛男
WU Tong;HUANG Yu;HOU Wanchao;XIA Jianli;LIU Chunming;LI Sainan(The Central Laboratory,Changchun Normal University,Changchun 130032)
出处
《分析科学学报》
CAS
CSCD
北大核心
2020年第3期353-358,共6页
Journal of Analytical Science
基金
国家自然科学基金(No.31870336,31670358)
吉林省科技发展计划(No.20180414028GH)
长春师范大学研究生教育创新计划项目(No.cscxy2018040)。
关键词
高速逆流色谱
超滤质谱
葛花
异黄酮
活性
HSCCC
Ultrafiltration-MS/MS
Puerariae Flos
Isoflavones
Activity
