不同产地赤芝药材的HPLC指纹图谱建立及聚类分析
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摘要
目的:建立赤芝药材的高效液相指纹图谱并进行聚类分析。方法:收集不同产地、不同批号的10批赤芝药材,以灵芝酸A、灵芝酸G为参照物,采用ZORBAX SB-C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相:乙腈-0.2%磷酸水,梯度洗脱;流速1.0 mL·min~(-1);检测波长254 nm。利用"中药色谱指纹图谱相似度评价系统2004A"建立指纹图谱,评价图谱的相似性,并进行聚类分析。结果:建立了10批赤芝药材的HPLC指纹图谱,共标定31个共有色谱峰,方法学考察结果符合指纹图谱的技术要求,相同产地的药材基本可以聚为一类。结论:该方法稳定、可靠、重复性好,为灵芝药材的质量评价提供了新的方法。
Objective:To establish an HPLC fingerprint of Ganoderma lucidum.Method:10 batches of G.lucidum from different origins and batches were collected,ganoderic acid A and ganoderic acid were used as reference materials,the HPLC method was used in the study with ZORBAX SB-C18(250 mm×4.6 mm,5 μm)column and mobile phase of acetonitrile-0.2% phosphoric acid water,the flow rate was 1.0 mL·min~(-1),and detection wavelength was 254 nm.The similarity of the obtained fingerprints were evaluated by the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2004 A.Results:The 10 batch of G.lucidum HPLC fingerprints were established,a total of 31 common peaks were calibrated,the method was validated.The medicinal materials of the same origin could be basically grouped into one class.Conclusion:The method is stable,reliable,reproducible,and provides a new method to evaluate the quality of G.lucidum.
Objective:To establish an HPLC fingerprint of Ganoderma lucidum.Method:10 batches of G.lucidum from different origins and batches were collected,ganoderic acid A and ganoderic acid were used as reference materials,the HPLC method was used in the study with ZORBAX SB-C18(250 mm×4.6 mm,5 μm)column and mobile phase of acetonitrile-0.2% phosphoric acid water,the flow rate was 1.0 mL·min~(-1),and detection wavelength was 254 nm.The similarity of the obtained fingerprints were evaluated by the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2004 A.Results:The 10 batch of G.lucidum HPLC fingerprints were established,a total of 31 common peaks were calibrated,the method was validated.The medicinal materials of the same origin could be basically grouped into one class.Conclusion:The method is stable,reliable,reproducible,and provides a new method to evaluate the quality of G.lucidum.
引文
[1] 江丽青.川北灵芝质量标准及其指纹图谱研究[D].四川:成都理工大学,2013.
[2] 赵如诗,姜慧洁,张晓静,等.灵芝三萜的高效液相色谱及薄层色谱指纹图谱研究[J].中华中医药杂志,2017,32(11):5167-5170.
[3] 张晓冬,陈龙,白玉,等.近红外光谱结合主成分分析和聚类分析鉴别炉甘石生品、伪品和炮制品[J].中国实验方剂学杂志,2018,24(12):1-8.
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[11] 何春龙,张屏,鞠爱华,等.灵芝超微粉、普通粉和灵芝提取物高效液相色谱指纹图谱的比较研究[J].中华中医药杂志,2017,32(2):575-578.
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[15] 黄必奎,陆兴毅,农凤鸣,等.广西百色地产灵芝FTIR特征图谱的系统聚类分析与主成分分析[J].中国民族民间医药,2010,19(5):24-26.
[16] CHAO W Z,TANG C H,ZHANG J S,et al.Development of a stable SCAR marker for rapid identification of Ganoderma lucidum Hunong 5 cultivar using DNA pooling method and inter-simple sequence repeat markers[J].J INTEGR AGR,2018,17(1):130-138.
[17] 冯柏康,李其凤,李卫民,等.HPLC指纹图谱结合系统聚类分析法对灵芝孢子油质量的研究[J].中国药房,2011,22(35):3326-3328.
[18] 贾红岩,王亚涛,张芝华,等.高效液相色谱法测定不同产地及品种灵芝三萜类成分的含量[J].微生物学通报,2017,44(1):238-244.
[2] 赵如诗,姜慧洁,张晓静,等.灵芝三萜的高效液相色谱及薄层色谱指纹图谱研究[J].中华中医药杂志,2017,32(11):5167-5170.
[3] 张晓冬,陈龙,白玉,等.近红外光谱结合主成分分析和聚类分析鉴别炉甘石生品、伪品和炮制品[J].中国实验方剂学杂志,2018,24(12):1-8.
[4] 张华锋,刘炯,张杰,等.基于聚类分析和典型相关分析的北柴胡保肝作用谱效关系研究[J].中草药,2013,44(19):2696-2702.
[5] 孟令文,田淋淋,李志成,等.长白山林下仿野生栽培紫芝主要活性成分研究[J].中国现代中药,2016,18(2):137-142.
[6] 赵维良,程夏倩,严爱娟,等.灵芝孢子油指纹图谱研究及其混淆品鉴别[J].中国现代应用药学,2017,34(7):1011-1014.
[7] WU W W,YE Z J,XU Y M,et al.Ergosta-7,22-diene-2β,3α,9α-triol(EGDT)from Ganoderma lucidum inhibits nasopharyngeal carcinoma cells by blocking EGFR signaling pathway[J].CHM,2018,10(1):27-33.
[8] 丁平,黄海波,邱金英,等.应用色谱技术区别赤芝和紫芝[J].华西药学杂志,2009,24(4):404-406
[9] 黄生权,魏刚,姚松君,等.赤芝、紫芝三萜类成分指纹图谱的优化与比较[J].华南理工大学学报(自然科学版),2010,38(8):121-124
[10] 郭晓蕾,翟旭峰,王怀豫,等.不同产地灵芝(赤芝)的HPLC指纹图谱评价[J].中华中医药学刊,2011,29(12):2782-2784.
[11] 何春龙,张屏,鞠爱华,等.灵芝超微粉、普通粉和灵芝提取物高效液相色谱指纹图谱的比较研究[J].中华中医药杂志,2017,32(2):575-578.
[12] LIANG J Y,ZHOU J H,ZHOU J P,et al.The Impact on Extraction Effect of Polysaccharide from Ganoderma lucidum by Different Pretreatment[J].Medicinal Plant,2018,9(3):14-16,20.
[13] 徐雪峰,闫浩,张玉.不同产地赤芝多糖和三萜含量的比较[J].海峡药学,2016,28(12):39-41.
[14] 孙小梅.灵芝子实体多糖色谱指纹分析方法的研究[D].无锡:江南大学,2014.
[15] 黄必奎,陆兴毅,农凤鸣,等.广西百色地产灵芝FTIR特征图谱的系统聚类分析与主成分分析[J].中国民族民间医药,2010,19(5):24-26.
[16] CHAO W Z,TANG C H,ZHANG J S,et al.Development of a stable SCAR marker for rapid identification of Ganoderma lucidum Hunong 5 cultivar using DNA pooling method and inter-simple sequence repeat markers[J].J INTEGR AGR,2018,17(1):130-138.
[17] 冯柏康,李其凤,李卫民,等.HPLC指纹图谱结合系统聚类分析法对灵芝孢子油质量的研究[J].中国药房,2011,22(35):3326-3328.
[18] 贾红岩,王亚涛,张芝华,等.高效液相色谱法测定不同产地及品种灵芝三萜类成分的含量[J].微生物学通报,2017,44(1):238-244.