防风薄层色谱指纹图谱的建立及其方法学研究

Fingerprint analysis of Saposhnikoviae Radix with high performance thin layer chromatography

目的建立防风高效薄层色谱指纹鉴别方法。方法以防风对照药材、升麻苷、5-O-甲基维斯阿米醇苷、紫花前胡苷、亥茅酚苷和升麻素为对照,采用单因素试验方法,对影响薄层色谱的各种因素进行系统考察,筛选出最佳的薄层色谱条件,并对不同批次的药材进行鉴别。结果筛选出最佳提取条件为10倍量85%的乙醇室温条件下超声提取20 min;薄层色谱条件为展开板HPTLC Silica G60(Merck),点样量5μl,条带宽10 mm,展开系统为展开剂Ⅰ:二氯甲烷-甲醇-甲酸(16∶4∶1),5 cm;展开剂Ⅱ:环己烷-乙酸乙酯(2∶1),8 cm,以10%硫酸乙醇溶液为显色剂,紫外灯365 nm下检视。所得供试品色谱中,在与对照品色谱图相应位置上,显示相同颜色的荧光条带。结论该方法简单、快速,通过建立薄层指纹条带,可有效地对防风真伪进行鉴别。

Objective To establish a high performance thin layer chromatography（ HPTLC） method for qualitative identification of Saposhnikoviae Radix（ SR）. Methods The SR reference substances,prim-O-glucosylcimifugin（ GC）,4＇-O-β-glucosyl-5-O-methylvisamminol（ GV）,nodakenin,sec-O-glucosylhamaudol（ GH） and cimifugin（ C） were used as the reference substances. The single-factor test was used. A variety of factors influencing TLC were systematically investigated to screen the best TLC conditions for identification of different batches of medicines. Results The best extraction method was 10 times the amount of 85% ethanol by ultrasonic extraction for 20 min at room temperature. The best TLC conditions were as follows： silica gel G60 plates,sample volume 5 μl,band width 10 mm,developing solvent Ⅰ（ dichloromethane-methanol-formic acid 16 ∶ 4 ∶ 1）,5 cm,developing solvent Ⅱ（ cyclohexane-ethyl acetate（ 2 ∶ 1）,8 cm,alcohol-sulfuric acid 9 ∶ 1 as chromogenic agent,detection under ultraviolet light at 365 nm. The HPTLC specific spectrums of SR showed the same fluorescence bands in the corresponding position of the reference substance. Conclusion This method is simple and fast,which can be effective to identify SR.