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用液质联用法联合生化分析法筛选连翘叶中α-葡萄糖苷酶和酪氨酸酶抑制作用的成分

Screening forα-glucosidase and tyrosinase inhibitors from Forsythia suspensa leaves by LC-MS technology combined biochemical analysis method
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摘要 目的建立一种液质结合生化分析法对连翘叶中具有α-葡萄糖苷酶和酪氨酸酶抑制作用的成分进行快速筛选。方法连翘叶提取液先由高效液相色谱进行分离,根据不同成分的出峰时间手动采集其中10个馏分,然后用分光光度法测定所采集的各组分对酪氨酸酶和α-葡萄糖苷酶的抑制率,最后用液质联用技术对所筛选的成分进行定性分析。结果本研究共筛选出对α-葡萄糖苷酶和酪氨酸酶具有抑制作用的成分各7种,并对其中5种成分进行了定性,馏分3为木通苯乙醇苷B或车前草苷B、馏分4为(+)-表儿茶酚-4-O-β-D-葡萄糖苷、馏分5为连翘苷、馏分6为连翘苷的同分异构体、馏分7为木通苯乙醇苷C。其中,馏分3、4、5这3种成分对2种酶均具有较强的抑制作用,馏分6对α-葡萄糖苷酶有较强的抑制作用。结论该方法操作简便、分离速度快、分析准确度好,可以很好地用于中药中其他活性成分的快速筛选。 Objective To build a liquid chromatography mass spectrometry technology combined biochemical analysis method for screeningα-glucosidase and tyrosinase inhibitors from Forsythia suspense leaves extract.Method Forsythia suspense leaves extract was injected into the high performance liquid chromatography system for the separation and 10 fractions were collected manually according to the peak time of each injection.The bioassays were determined by spectrophotometric method,the high performance liquid chromatography mass spectrometry was used for the characterization.Results A total of 7 fractions were screened from forsythia leaf extract which had inhibitory effects onα-glucosidase and tyrosinase inhibitors,5 compounds were characterized,and the qualitative results showed that fraction 3 was calceolarioside B or plantainoside B,fraction 4 was(+)-epicatechol-4-O-β-D-glucoside,fraction 5 was phillyrin,fraction 6 was the isomeride of phillyrin,and fraction 7 was calceolarioside C.Fraction 3,4 and 5 showed inhibitory effect on both tyrosinase andα-glucosidase,fraction 6 showed inhibitory effect onα-glucosidase only.Conclusion The test results showed that the proposed method was a simple,effective,rapid method for drug discovery in natural products,and it can be extended to screen other bioactive compounds from natural products and o ther complex samples.
作者 徐敬朴 陶兴隆 郑丽亚 李德强 XU Jing-pu;TAO Xing-long;ZHENG Li-ya;Li De-qiang(Department of Pharmacy,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)
出处 《中国临床药理学杂志》 CAS CSCD 北大核心 2024年第23期3453-3457,共5页 The Chinese Journal of Clinical Pharmacology
基金 河北省自然科学基金资助项目(H202306050) 河北省中医药管理局课题基金资助项目(2021144)。
关键词 连翘 液质联用法 生化分析法 Α-葡萄糖苷酶 酪氨酸酶 Forsythia suspensa LC-MS technology biochemical analysis method α-glucosidase tyrosinase
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