基于指纹图谱结合多指标测定的玄参药材等级质量研究
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摘要
目的建立玄参药材的HPLC指纹图谱,并同时进行多成分含量测定,为玄参药材等级质量标准的制定提供方法和依据。方法采用HPLC-DAD法,Elipse XDB-C18(250 mm×4.6 mm,5μm)色谱柱,柱温25℃,以乙腈(A)-0.03%磷酸水溶液(B)为流动相进行梯度洗脱,体积流量1.0 mL/min,进样量10μL,指纹图谱检测波长207 nm,含量测定的检测波长为哈巴苷210 nm、哈巴俄苷280 nm、肉桂酸264 nm。获取30批不同等级玄参药材的指纹图谱,采用中药色谱指纹图谱相似度评价系统(2012版)进行分析,同时对3种主要化学成分进行含量测定。结果指纹图谱分析共指认7个共有峰,相对保留时间RSD值均小于2.0%,相对峰面积RSD值差异较大,提示玄参药材中的主要化学成分稳定存在,但含量差异较大;相似度分析中,约有5%的指纹图谱与对照药材的相似度低于0.7,12.5%的相似度在0.7～0.8,40%处于0.8～0.9,42.5%大于0.9,提示不同产地玄参药材的质量差异较大;不同批次玄参药材中3种化学成分含量差异较大,质量分数分别为哈巴苷0.18%～2.89%、哈巴俄苷0.01%～0.35%、肉桂酸0.01%～0.24%。结论建议在原有等级划分标准的基础上,增加指纹图谱和主要化学成分的含量测定两项指标。
Objective In order to provide a scientific basis for the grade quality standard criterion of Scrophulariae Radix, a HPLC-DAD method was established to acquire fingerprint and detect the content of multiple components. Methods The chromatographic separate was achieved on Elipse XDB-C18(250 mm × 4.6 mm, 5 μm) column, with the temperature of 25 ℃,using acetonitrile(A)-0.03% phosphoric acid water(B) as the mobile phase gradient elution, a flow rate of 1.0 mL/min, and the detection wavelength was set at 207 nm for fingerprint, 210 nm for Harpagide, 280 nm for harpagoside, and 264 nm for cinnamic acid. The injection volume was 10 μL. The fingerprints of different grades of Scrophulariae Radix from 30 batches were evaluated with a chromatographic fingerprint similarity evaluation system(version 2012), and the content of harpagide, harpagoside, and cinnamic acid was also determined at the same time. Results There were seven common peaks in the fingerprints, the RSD values of relative retention time were lower than 2.0% and those of relative peak area were quite different, which indicated that the main chemical compounds can exist stably in Scrophulariae Radix with different content. Compared with control, the results of fingerprint similarity were presented as follows: 5% under 0.7, 12.5% arranged 0.7 to 0.8, 40% between 0.8 and 0.9, and another 42.5% exceed0.9, demonstrated that there was qualitative difference in various batches of Scrophulariae Radix. Also, the quantitative analysis of multi-index showed the differences in three main compounds, the content respectively was 0.18%—2.89% in harpagide, 0.01%—0.35% in harpagoside, and 0.01%—0.24% in cinnamic acid. Conclusion Suggestions were provided in the formulation of new grade quality standard, such as adding the fingerprints and multi-index detection of main chemical components based on the original criterion of classification.
Objective In order to provide a scientific basis for the grade quality standard criterion of Scrophulariae Radix, a HPLC-DAD method was established to acquire fingerprint and detect the content of multiple components. Methods The chromatographic separate was achieved on Elipse XDB-C18(250 mm × 4.6 mm, 5 μm) column, with the temperature of 25 ℃,using acetonitrile(A)-0.03% phosphoric acid water(B) as the mobile phase gradient elution, a flow rate of 1.0 mL/min, and the detection wavelength was set at 207 nm for fingerprint, 210 nm for Harpagide, 280 nm for harpagoside, and 264 nm for cinnamic acid. The injection volume was 10 μL. The fingerprints of different grades of Scrophulariae Radix from 30 batches were evaluated with a chromatographic fingerprint similarity evaluation system(version 2012), and the content of harpagide, harpagoside, and cinnamic acid was also determined at the same time. Results There were seven common peaks in the fingerprints, the RSD values of relative retention time were lower than 2.0% and those of relative peak area were quite different, which indicated that the main chemical compounds can exist stably in Scrophulariae Radix with different content. Compared with control, the results of fingerprint similarity were presented as follows: 5% under 0.7, 12.5% arranged 0.7 to 0.8, 40% between 0.8 and 0.9, and another 42.5% exceed0.9, demonstrated that there was qualitative difference in various batches of Scrophulariae Radix. Also, the quantitative analysis of multi-index showed the differences in three main compounds, the content respectively was 0.18%—2.89% in harpagide, 0.01%—0.35% in harpagoside, and 0.01%—0.24% in cinnamic acid. Conclusion Suggestions were provided in the formulation of new grade quality standard, such as adding the fingerprints and multi-index detection of main chemical components based on the original criterion of classification.
引文
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[2]明·缪希雍.神农本草经疏[M].北京:中国中医药出版社,1997.
[3]刘树民,张宁,于卉,等.玄参性味的本草考证[J].中华中医药杂志,2017,32(2):429-431.
[4]张刘强,李医明.近10年玄参属植物化学成分和药理作用研究进展[J].中草药,2011,42(11):2360-2368.
[5]刘宾,梁晨,徐思思,等.不同产地玄参饮片哈巴苷与哈巴俄苷含量比较研究[J].时珍国医国药,2014,25(3):555-556.
[6]七十六种药材商品规格标准[S].1984.
[7]杨俊杰,李林,季德,等.中药材产地加工与炮制一体化的历史沿革与现代研究探讨[J].中草药,2016,47(15):2751-2757.
[8]杨光,曾燕,郭兰萍,等.中药材商品规格等级标准研究现状及几个关键问题的商榷[J].中国中药杂志,2014,39(9):1733-1738.