不同来源金线莲的HPLC指纹图谱
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摘要
目的建立不同来源金线莲(组培、栽培、野生)HPLC指纹图谱,并比较不同来源金线莲及其近缘植物整体的差异,为金线莲质量评价及"真伪"的鉴别提供方法。方法采用Agilent TC-C_(18)色谱柱(250 mm×4.6 mm,5μm),乙腈-水为流动相进行梯度洗脱,流速为1 mL·min~(-1),检测波长为260 nm,柱温30℃,采用相似度评价和聚类分析,分别对37批不同来源金线莲(组培、栽培、野生)及11批近缘植物(台湾金线莲、血叶兰)指纹图谱进行分析。结果分别建立了不同来源即组培、栽培和野生金线莲HPLC指纹图谱,分别确定了11、19和12个共有峰,指认了5、5和2个共有峰,不同来源金线莲批次内相似度较高,但批次间化学成分存在差异。台湾金线莲与野生金线莲典型指纹图谱相似度为0.6～0.739,不能很好区分;血叶兰与野生金线莲典型指纹图谱明显不同,可用于鉴别。结论建立的不同来源金线莲的HPLC指纹图谱精密度、重复性、稳定性良好,为金线莲整体质量评价及来源的鉴别提供参考。
Objective To develop the HPLC fingerprints of different sources of Anoectochilus roxburghii(tissue culture,cultivated and wild)and to apply for the identification of different types of A.roxburghii and its relative plants.The finding also provided an effective analysis method for quality management of A.roxburghii and for authentification of flake product.Methods The HPLC fingerprint was performed on an Agilent TC-C_(18) column(250 mm×4.6 mm,5 μm)with a mobile phase of water-acetonitrile in gradient elution.The flow rate was 1 mL·min~(-1) and column temperature was set at 30 ℃.The detection wavelength was set at 260 nm.The similarity and cluster analysis were used to evaluate the fingerprints of 37 batches of A.roxburghii(tissue culture,cultivated and wild)and 11 batches of the allied plants(Anoectochilus formosanus and Ludisia discolor).Results The general chromatographic fingerprints for tissue culture,cultivated and wild A.roxburghii were established with 11,19,12 common fingerprint peaks,and 5,5,2 main compounds peaks were identified,respectively.The similarity was high among the batches of different sources of A.roxburghii,but there were significant differences in chemical composition between its batches.The similarity between the fingerprint of wild A.roxburghii and A.formosanus was 0.6 to 0.739,which could not be distinguished.However,there was obvious differences between the fingerprint of wild A.roxburghii and L.discolor,which could be used for identification.Conclusion The established HPLC fingerprints of different sources of A.roxburghii have a good precision,repeatability,stability,providing a reference for the objective and effective identification and quality evaluation of A.roxburghii.
Objective To develop the HPLC fingerprints of different sources of Anoectochilus roxburghii(tissue culture,cultivated and wild)and to apply for the identification of different types of A.roxburghii and its relative plants.The finding also provided an effective analysis method for quality management of A.roxburghii and for authentification of flake product.Methods The HPLC fingerprint was performed on an Agilent TC-C_(18) column(250 mm×4.6 mm,5 μm)with a mobile phase of water-acetonitrile in gradient elution.The flow rate was 1 mL·min~(-1) and column temperature was set at 30 ℃.The detection wavelength was set at 260 nm.The similarity and cluster analysis were used to evaluate the fingerprints of 37 batches of A.roxburghii(tissue culture,cultivated and wild)and 11 batches of the allied plants(Anoectochilus formosanus and Ludisia discolor).Results The general chromatographic fingerprints for tissue culture,cultivated and wild A.roxburghii were established with 11,19,12 common fingerprint peaks,and 5,5,2 main compounds peaks were identified,respectively.The similarity was high among the batches of different sources of A.roxburghii,but there were significant differences in chemical composition between its batches.The similarity between the fingerprint of wild A.roxburghii and A.formosanus was 0.6 to 0.739,which could not be distinguished.However,there was obvious differences between the fingerprint of wild A.roxburghii and L.discolor,which could be used for identification.Conclusion The established HPLC fingerprints of different sources of A.roxburghii have a good precision,repeatability,stability,providing a reference for the objective and effective identification and quality evaluation of A.roxburghii.
引文
[1] 福建省食品药品监督管理局.福建中药材标准[M].福州:海风出版社,2006:154-157.
[2] ZHANG J G,LIU Q,LIU Z L,et al.Antihyperglycemic activity of Anoectochilus Roxburghii Polysaccharose in diabetic mice induced by high-fat diet and streptozotocin [J].Journal of Ethnopharmacology,2015,164:180-185.
[3] 陈学香,夏向南.金线莲治疗高龄老年高尿酸血症疗效观察[J].中华保健医学杂志,2010,12(4):308-309.
[4] 刘政芳,李芹.复方金线莲口服液联合恩替卡韦治疗慢性乙型肝炎30例临床观察[J].福建中医药,2008,39(5):3-4.
[5] YE Shenyi,SHAO Qingsong,ZHANG Ailian.Anoectochilus roxburghii:a review of its phytochemistry,pharmacology,and clinical applications[J].Journal of Ethnopharmacology,2017,209:184-202.
[6] 刘知远,沈廷明,吴仲玉.RP-HPLC法同时测定福建产金线莲中槲皮素、山柰素、异鼠李素的量[J].中草药,2015,46(3):432-434.
[7] 何春年.福建金线莲的化学成分研究[D].北京:中国协和医科大学,2004.
[8] 藏蔓,卢敬光,谢启师,等.基于UHPLC-TOF-MS技术建立金线莲的指纹图谱及金线莲苷的含量测定方法[J].中国实验方剂学杂志,2018,24(1):31-37.
[9] 李瑞姿,林军,汪厦霞,等.基于不同栽培阶段金线莲的代谢组学分析[J].中国中药杂志,2017,42(23):4624-4630.
[10] 蔡金艳,张勇慧,吴继洲.药王金线莲挥发油及石油醚提取物的气相色谱-质谱-数据系统联用测定[J].时珍国医国药,2008,19(7):1550-1552.
[11] 林平,华碧春,黄智锋.金线莲的质量标准探讨[J].福建中医药大学学报,2012,22(5):40-42.
[12] 王晓雪.福建金线莲粗提物多组分的快速质谱分析方法及NMR/RRLC-MS相关谱分析方法研究[D].北京:北京协和医学院,2011.
[13] 黄瑞平,黄颖桢,陈菁瑛,等.不同月龄金线莲多糖和总黄酮含量的比较[J].热带生物学报,2012,3(2):174-176.
[14] 王澄林.近红外光谱技术在金线莲定性定量分析中的应用研究[D].福州:福建中医药大学,2017.
[15] 黄颖桢,陈菁瑛,赵云青,等.金线莲遗传多样性的ISSR分析[J].中草药,2014,45(15):2230-2234.
[16] 李雪娇,李姣姣,王淼,等.参苓白术散GC指纹图谱检测方法[J].沈阳药科大学学报,2019,36(1):61-65.
[17] 吴萍萍,黄丽英.金线莲HPLC指纹图谱研究[J].中草药,2015,46(13):1975-1979.
[18] 陈莹,黄锦芳,林娜,等.HPLC法同时测定金线莲中6种成分[J].中成药,2018,40(10):2222-2227.
[2] ZHANG J G,LIU Q,LIU Z L,et al.Antihyperglycemic activity of Anoectochilus Roxburghii Polysaccharose in diabetic mice induced by high-fat diet and streptozotocin [J].Journal of Ethnopharmacology,2015,164:180-185.
[3] 陈学香,夏向南.金线莲治疗高龄老年高尿酸血症疗效观察[J].中华保健医学杂志,2010,12(4):308-309.
[4] 刘政芳,李芹.复方金线莲口服液联合恩替卡韦治疗慢性乙型肝炎30例临床观察[J].福建中医药,2008,39(5):3-4.
[5] YE Shenyi,SHAO Qingsong,ZHANG Ailian.Anoectochilus roxburghii:a review of its phytochemistry,pharmacology,and clinical applications[J].Journal of Ethnopharmacology,2017,209:184-202.
[6] 刘知远,沈廷明,吴仲玉.RP-HPLC法同时测定福建产金线莲中槲皮素、山柰素、异鼠李素的量[J].中草药,2015,46(3):432-434.
[7] 何春年.福建金线莲的化学成分研究[D].北京:中国协和医科大学,2004.
[8] 藏蔓,卢敬光,谢启师,等.基于UHPLC-TOF-MS技术建立金线莲的指纹图谱及金线莲苷的含量测定方法[J].中国实验方剂学杂志,2018,24(1):31-37.
[9] 李瑞姿,林军,汪厦霞,等.基于不同栽培阶段金线莲的代谢组学分析[J].中国中药杂志,2017,42(23):4624-4630.
[10] 蔡金艳,张勇慧,吴继洲.药王金线莲挥发油及石油醚提取物的气相色谱-质谱-数据系统联用测定[J].时珍国医国药,2008,19(7):1550-1552.
[11] 林平,华碧春,黄智锋.金线莲的质量标准探讨[J].福建中医药大学学报,2012,22(5):40-42.
[12] 王晓雪.福建金线莲粗提物多组分的快速质谱分析方法及NMR/RRLC-MS相关谱分析方法研究[D].北京:北京协和医学院,2011.
[13] 黄瑞平,黄颖桢,陈菁瑛,等.不同月龄金线莲多糖和总黄酮含量的比较[J].热带生物学报,2012,3(2):174-176.
[14] 王澄林.近红外光谱技术在金线莲定性定量分析中的应用研究[D].福州:福建中医药大学,2017.
[15] 黄颖桢,陈菁瑛,赵云青,等.金线莲遗传多样性的ISSR分析[J].中草药,2014,45(15):2230-2234.
[16] 李雪娇,李姣姣,王淼,等.参苓白术散GC指纹图谱检测方法[J].沈阳药科大学学报,2019,36(1):61-65.
[17] 吴萍萍,黄丽英.金线莲HPLC指纹图谱研究[J].中草药,2015,46(13):1975-1979.
[18] 陈莹,黄锦芳,林娜,等.HPLC法同时测定金线莲中6种成分[J].中成药,2018,40(10):2222-2227.
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