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胡芦巴种子70%醇提物薄层色谱和高效液相色谱特征图谱的建立

Establishment of thin layer chromatography and high performance liquid chromatography characteristic chromatograms of 70%ethanol extract from seeds of fenugreek
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摘要 目的:建立胡芦巴种子70%乙醇提取物的薄层色谱(TLC)和高效液相色谱(HPLC)特征图谱,同时考察11批胡芦巴种子醇提物特征图谱的相似性和差异性。方法:①采用G型和HF_(254)型薄层层析硅胶板,以正丁醇-乙醇-水(2:6:2)为展开剂,分别喷洒碘化铋钾-三氯化铁和10%硫酸-乙醇2种显色剂,在日光、紫外波长254和365nm下检视色谱斑点,建立胡芦巴70%醇提物的TLC特征图谱。②采用SupersilODS-B色谱柱(250mm×4.6mm,5μm),以0.1%磷酸溶液-乙腈为流动相进行梯度洗脱,流速0.7mL·min^(-1),柱温30℃,检测波长203nm,建立胡芦巴70%醇提物的HPLC特征图谱。结果:TLC特征图谱显示该醇提物有8个明显斑点,其中含抗糖尿病的活性成分胡芦巴碱和薯蓣皂苷。HPLC特征图谱显示该醇提物含有29个共有色谱峰,其中指认出14种色谱峰的化学成分。相似度评价表明11批胡芦巴醇提物相似度>0.9。聚类分析表明不同产地的胡芦巴药材之间存在一定的差异性,其中安徽与河南产地的胡芦巴药材相似度较高。结论:该方法稳定性和重复性良好,而且具有较强的特征性,可用于胡芦巴70%醇提物的质量控制。 Objective:To establish thin layer chromatography(TLC)and high performance liquid chroma-tography(HPLC)characteristic chromatography methods of 70%ethanol extract from seeds of fenugreek,and to investigate the similarities and differences among eleven batches of fenugreek extract.Methods:G and HF_(254) silica gel plates were used for TLC,using a mixture of n-butanol-ethanol-water(2:6:2)as developer,bismuth potassium iodide-ferric chlorideand and 10%sulphuric acid-ethanol as color developing reagents.The sunlight and ultraviolet wavelength at 254 nm and 365 nm were used to identify the spots on TLC plats.②A Supersil ODS-B column(250 mm×4.6 mm,5μm)was used for HPLC chromatography.The mobile phase was 0.1%phosphoric acid-acetonitrile with a gradient elution.The flow rate was 0.7 mL·min^(-1),the column temperature was 30 C,and the detection wavelength was 203 nm.Results:The TLC chromatogram of 70%ethanol extract from fenugreek indicated that there were eight spots containing trigonelline and dioscin as anti-diabetic active components. InHPLC chromatogram,there were twenty-nine common peaks in eleven batches of fenugreek,and fourteen componentswere identified. The similarity of the eleven batches of 70% ethanol extract were more than 0. 9. The cluster analysisshowed that there was variability among the samples of fenugreek from different origins. However,there are highsimilarity between Anhui and Henan provinces. Conclusion: These developed methods are reproducible and reliable.The two chromatograms showed strong characteristics,which can be used to control the quality of 70% ethanolextractof fenugreek.
作者 孟凡恒 朱帅鸣 骆芙瑶 徐锐 单俊杰 MENG Fan-heng;ZHU Shuai-ming;LUO Fu-yao;XU Rui;SHAN Jun-jie(College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210046,China;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)
出处 《中国新药杂志》 CAS CSCD 北大核心 2023年第7期752-760,共9页 Chinese Journal of New Drugs
关键词 胡芦巴 醇提物 薄层色谱 高效液相色谱 特征图谱 fenugreek ethanol-extract thin layer chromatography high performance liquid chromatography characteristic chromatogram
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