应用赤芍对照提取物对赤芍样品的指纹图谱分析
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摘要
目的使用赤芍对照提取物(CS ERS)对市场上的赤芍药材进行定性鉴别及定量分析。方法使用薄层色谱法、硅胶薄层板,以甲苯-乙酸乙酯-甲醇-水-冰醋酸(10∶7∶5∶0.5∶0.1)为展开剂,对赤芍样品进行定性鉴别。采用Waters Symmetry C_(18) column(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.1%磷酸溶液,梯度洗脱;流速0.8 mL·min~(-1),柱温25℃,检测波长230 nm,进样量10μL。用芍药苷对照品外标法测定赤芍中芍药苷的含量,用没食子酸、儿茶素、没食子酰甲酯、芍药内酯苷、五没食子酰葡萄糖(5-GG)、苯甲酸、苯甲酰芍药苷和丹皮酚鉴别赤芍中的成分。结果薄层色谱显示CS ERS与赤芍药材的芍药苷在同一Rf值上显示相同的斑点,在整体上具有一致性,CS ERS能有效鉴别出质量好的赤芍药材;高效液相显示23个赤芍样品中有19个赤芍样品的芍药苷含量≥1.8%;不同赤芍样品中的化学成分比例不同,可以分成4类。结论赤芍样品种类多样,利用CS ERS能有效鉴别出质量好的一类赤芍药材。应用中药对照提取物用于中药质量符合中药整体性和复杂性的特点,更适合中药整体质量控制的需求。
Objective Our study assessed the application of CS ERS in TLC identification and HPLC determination of Paeoniae radix rubra. Methods TLC was conducted on pre-coated silica gel 60 performance TLC plate,developing with the solvent system of toluene,ethyl acetate,methanol,water,glacial acetic acid in the proportion of 10∶7∶5∶0.5∶0.1. The samples were separated on a Waters Symmetry C_(18) column(250 mm×4.6 mm,5 μm)using acetonitrile and 0.1 % phosphoric solution as mobile phase in gradient elution. The flow rate was 0.8 mL·min~(-1) and the column temperature was 25 ℃. The detection wavelength was set at 230 nm and 10 μL of the sample was injected for analysis. The content of paeoniflorin in Paeoniae radix rubra was determined by external standard method of paeoniflorin reference substance and the components in Paeoniae radix rubra were identified by gallic acid,catechin, methyl gallate, albiflorin, benzoic acid, benzoylpaeoniflorin and paeonol. Results CS ERS demonstrated the same TLC pattern as Paeoniae radix rubra samples in which Paeoniflorin could be observed with the same Rf value. CS ERS and Paeoniae radix rubra are consistent on the whole. The HPLC results show that the paeoniflorin contents of 19 out of 23 batches of Paeoniae radix rubra samples are higher than 1.8% and CS ERS can be used for effectively identifying a class of Paeoniae radix rubra with good quality. Paeoniae radix rubra samples with different chemical compositions proportions can be divided into four kinds. Conclusion CS ERS can be used to identify Paeoniae radix rubra with good quality. Extract reference substance was consistent with the integrity and complexity of traditional Chinese medicine and was suitable for quality control. Using CS ERS for TLC identification and HPLC determination was convenient,effective and affordable.
Objective Our study assessed the application of CS ERS in TLC identification and HPLC determination of Paeoniae radix rubra. Methods TLC was conducted on pre-coated silica gel 60 performance TLC plate,developing with the solvent system of toluene,ethyl acetate,methanol,water,glacial acetic acid in the proportion of 10∶7∶5∶0.5∶0.1. The samples were separated on a Waters Symmetry C_(18) column(250 mm×4.6 mm,5 μm)using acetonitrile and 0.1 % phosphoric solution as mobile phase in gradient elution. The flow rate was 0.8 mL·min~(-1) and the column temperature was 25 ℃. The detection wavelength was set at 230 nm and 10 μL of the sample was injected for analysis. The content of paeoniflorin in Paeoniae radix rubra was determined by external standard method of paeoniflorin reference substance and the components in Paeoniae radix rubra were identified by gallic acid,catechin, methyl gallate, albiflorin, benzoic acid, benzoylpaeoniflorin and paeonol. Results CS ERS demonstrated the same TLC pattern as Paeoniae radix rubra samples in which Paeoniflorin could be observed with the same Rf value. CS ERS and Paeoniae radix rubra are consistent on the whole. The HPLC results show that the paeoniflorin contents of 19 out of 23 batches of Paeoniae radix rubra samples are higher than 1.8% and CS ERS can be used for effectively identifying a class of Paeoniae radix rubra with good quality. Paeoniae radix rubra samples with different chemical compositions proportions can be divided into four kinds. Conclusion CS ERS can be used to identify Paeoniae radix rubra with good quality. Extract reference substance was consistent with the integrity and complexity of traditional Chinese medicine and was suitable for quality control. Using CS ERS for TLC identification and HPLC determination was convenient,effective and affordable.
引文
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[2]杨柳,许舜军,吴金雄,等.白芍、赤芍的比较研究概况[J].中药新药与临床药理,2011:22(5):577-580.
[3]江苏新医学院.中药大辞典(1997版)[M].上海:上海科技出版社,2003:706-709,1093-1095.
[4]阮金兰,赵钟祥,曾庆忠.赤芍化学成分和药理作用的研究进展[J].中国药理学通报,2003,19(9):965-970.
[5]王瑞.川赤芍化学成分与芍药质量控制方法研究[D].沈阳:沈阳药科大学,2005.
[6]谢培山,颜玉贞.中药色谱指纹图谱精细分析图集[M].福州:福建科学技术出版社,2015:52-61.
[7]王峥涛,谢培山.中药材质量专论[M].上海:上海科学技术出版社,2013:62-72.
[8]周红涛,骆亦奇,胡世林,等.赤芍与白芍的化学成分含量比较研究[J].中国药学杂志,2013,38(9):654.
[9] XIE P S,MA S C,TU P F,et al. The prospect of application of extractive reference substance of Chinese Herbal Medicines[J].Chinese Medicine,2013,4(4):125-136.
[10]翟海斌,欧丹林,程翼宇.中药提取物质量控制的一种新方法探讨[J].中国药学杂志,2006,31(1):57-60.
[11]谢培山.中药色谱指纹图谱质量控制模式的研究和应用—若干实际性问题的探讨(一)[J].世界科学技术,2001,3(3):18-23.