Rojsanga Piyanuch, Gritsanapan Wandee, Suntornsuk Leena
Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University, Ratchatawi, Bangkok, Thailand.
Med Princ Pract. 2006;15(5):373-8. doi: 10.1159/000094272.
To develop the optimal extraction procedure (i.e. maceration, percolation or Soxhlet extraction) and thin- layer chromatographic (TLC)-densitometric method for the determination of berberine content of Coscinium fenestratum.
Maceration, percolation and Soxhlet extraction techniques were used to extract alkaloids from dried stems of C. fenestratum. The solvents used were 50 and 80% ethanol. Crude extracts and berberine content recovered from the TLC fingerprint were evaluated for chemical components of each extraction method. Precoated silica gel GF(254) plates were used as stationary phase while butanol:glacial acetic acid:water (14:3:4) was used as a mobile phase. Detection and quantitation of berberine were performed by densitometry at the wavelength of 415 nm over the linearity range of 240-840 ng (r(2) = 0.9982). The relative standard deviations from intraday and interday precisions were less than 4.13%.
The recovery of standard berberine was 97.58-98.71% (%RSD = 3.85), and the limit of detection and quantitation were 25 and 50 ng/spot, respectively. Eighty percent ethanol gave a higher content of berberine than 50% ethanol. Berberine contents from maceration, percolation and Soxhlet extraction with 80% ethanol were 3.37+/- 0.30, 3.08+/- 0.38 and 2.67+/- 0.27% w/w, respectively.
The TLC-densitometric method was simple, accurate and precise for quantitating berberine in the stem extract of C. fenestratum. Maceration with 80% ethanol gave the highest content of berberine in the extract. TLC of the extracts from different methods showed a similar pattern.
开发用于测定藤黄连中黄连素含量的最佳提取方法（即浸渍法、渗漉法或索氏提取法）及薄层色谱（TLC）-密度测定法。
采用浸渍法、渗漉法和索氏提取法从藤黄连干燥茎中提取生物碱。所用溶剂为50%和80%乙醇。对从TLC指纹图谱中回收的粗提取物和黄连素含量进行评估，以分析每种提取方法的化学成分。采用预涂硅胶GF(254)板作为固定相，以正丁醇：冰醋酸：水（14:3:4）作为流动相。在415 nm波长下，于240 - 840 ng的线性范围内通过密度测定法对黄连素进行检测和定量（r(2) = 0.9982）。日内和日间精密度的相对标准偏差均小于4.13%。
标准黄连素的回收率为97.58 - 98.71%（%RSD = 3.85），检测限和定量限分别为25和50 ng/斑点。80%乙醇提取得到的黄连素含量高于50%乙醇。用80%乙醇通过浸渍法、渗漉法和索氏提取法得到的黄连素含量分别为3.37±0.30、3.08±0.38和2.67±0.27%（w/w）。
TLC-密度测定法用于定量藤黄连茎提取物中的黄连素简单、准确且精密。用80%乙醇浸渍法得到的提取物中黄连素含量最高。不同方法提取物的TLC显示出相似的图谱。
