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高效薄层色谱法及荧光密度计法测定生物样品中的小檗碱

[Determination of berberine in biological specimen by high performance TLC and fluoro-densitometric method].

作者信息

Shen Z F, Xie M Z

机构信息

Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing.

出版信息

Yao Xue Xue Bao. 1993;28(7):532-6.

PMID:8285056
Abstract

In this paper, a simple and specific high performance TLC and fluoro-densitometric method for separating and determining berberine in biological specimen was developed. Two hundred microliters of plasma or tissue homogenate mixed with 20 microliters of sodium lauryl sulfate (18%) and quinidine (300 ng, as internal standard) were extracted with 1 ml of chloroform. The developing solvent consisted of ethyl acetate-methyl acetate-methanol-water (5.4:4.6:1.2:1.0). The determination was carried out with a Shimadzu TLC-Scanner CS-910. Berberine spots were measured using an excitation wavelength of 350 nm and an emission wavelength of 550 nm, while using 350 nm and 450 nm for quinidine spots as the excitation and emission wavelength respectively. The average recoveries were 100.3% from plasma and 103.8% from tissue homogenate. The berberine levels in plasma and in tissues were compared in normal mice after oral administration of berberine (100 mg/kg) or the powder of Coptis chinensis (2 g/kg) containing the same amount of berberine. Simultaneously, the hypoglycemic effect of berberine and that of the powder of Coptis chinensis were measured in these mice. Results indicate that the concentrations of berberine in plasma and tissues given the powder of Coptis chinensis orally were higher than those given berberine and that the hypoglycemic effect of the powder was also stronger than that of berberine. Evidently, the changes of the blood glucose level and the level of berberine in plasma showed an opposite relationship.

摘要

本文建立了一种简单、特异的高效薄层色谱和荧光密度法,用于生物样品中黄连素的分离与测定。取200微升血浆或组织匀浆,加入20微升十二烷基硫酸钠(18%)和奎尼丁(300纳克,作为内标),用1毫升氯仿萃取。展开剂由乙酸乙酯 - 乙酸甲酯 - 甲醇 - 水(5.4:4.6:1.2:1.0)组成。测定采用岛津TLC - Scanner CS - 910进行。黄连素斑点的测定,激发波长为350纳米,发射波长为550纳米,而奎尼丁斑点的测定,激发波长和发射波长分别为350纳米和450纳米。血浆中的平均回收率为100.3%,组织匀浆中的平均回收率为103.8%。在正常小鼠口服黄连素(100毫克/千克)或含等量黄连素的黄连粉末(2克/千克)后,比较血浆和组织中的黄连素水平。同时,测定这些小鼠中黄连素和黄连粉末的降血糖作用。结果表明,口服黄连粉末后,血浆和组织中黄连素的浓度高于口服黄连素的情况,且粉末的降血糖作用也强于黄连素。显然,血糖水平的变化与血浆中黄连素水平呈相反关系。

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