夏天无HPLC指纹图谱及多指标含量测定研究
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摘要
目的建立夏天无HPLC指纹图谱及多指标成分测定方法,为夏天无药材质量标准提升提供科学依据。方法采用HPLC法,Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱;以乙腈-0.2%冰醋酸溶液(三乙胺调pH 6.0)为流动相,梯度洗脱;体积流量1.0 mL/min;柱温35℃;进样量10μL;检测波长280 nm;建立15批夏天无药材的指纹图谱,用中药色谱指纹图谱相似度评价系统软件(2012A版)进行评价,聚类分析和主成分分析将15批药材分为2类,同时测定原阿片碱、黄藤素、荷包牡丹碱和延胡索乙素4种成分的含量。结果建立了夏天无药材的HPLC指纹图谱,共标定了10个共有峰;原阿片碱、黄藤素、荷包牡丹碱和延胡索乙素线性关系良好,r均大于0.999 9,平均回收率分别为97.12%、100.09%、98.53%、99.71%。结论 HPLC指纹图谱结合多指标成分测定可为夏天无药材质量评价提供参考。
Objective To establish the HPLC fingerprint and multicomponents determination of Corydalis Decumbentis Rhizoma, and provide a scientific basis for the improvement of its quality standards. Methods The separation was performed on a chromatographic Diamonsil C18 column(250 mm × 4.6 mm, 5 μm), with acetonitrile-0.2% acetic acid(adjusting pH to 6.0 with triethylamine) as the mobile phase for gradient elution. Volume flow rate was 1.0 m L/min. Column temperature was 35 ℃. Injection was 10 μL and the detection wavelength was 280 nm. The fingerprints of 15 batches of Corydalis Decumbentis Rhizoma were established and evaluated by the similarity evaluation system of TCM(version 2012 A), which were divided into two categories by clustering analysis and principal component analysis.Meanwhile, the content of protopine, palmatine, bicuculline and tetrahydropalmatine was determinated. Results The fingerprint of Corydalis Decumbentis Rhizoma was established. There were 10 common peaks in the fingerprint. Protopine, palmatine, bicuculline and dl-tetrahydropalmatine were separated with good linearity relationships(r > 0.999 9). The average recovery rates of the investigated compounds were 97.12%, 100.09%, 98.53%, and 99.71%, respectively. Conclusion HPLC fingerprint combined with multicomponents determination can provide the reference for the quality evaluation of Corydalis Decumbentis Rhizoma.
Objective To establish the HPLC fingerprint and multicomponents determination of Corydalis Decumbentis Rhizoma, and provide a scientific basis for the improvement of its quality standards. Methods The separation was performed on a chromatographic Diamonsil C18 column(250 mm × 4.6 mm, 5 μm), with acetonitrile-0.2% acetic acid(adjusting pH to 6.0 with triethylamine) as the mobile phase for gradient elution. Volume flow rate was 1.0 m L/min. Column temperature was 35 ℃. Injection was 10 μL and the detection wavelength was 280 nm. The fingerprints of 15 batches of Corydalis Decumbentis Rhizoma were established and evaluated by the similarity evaluation system of TCM(version 2012 A), which were divided into two categories by clustering analysis and principal component analysis.Meanwhile, the content of protopine, palmatine, bicuculline and tetrahydropalmatine was determinated. Results The fingerprint of Corydalis Decumbentis Rhizoma was established. There were 10 common peaks in the fingerprint. Protopine, palmatine, bicuculline and dl-tetrahydropalmatine were separated with good linearity relationships(r > 0.999 9). The average recovery rates of the investigated compounds were 97.12%, 100.09%, 98.53%, and 99.71%, respectively. Conclusion HPLC fingerprint combined with multicomponents determination can provide the reference for the quality evaluation of Corydalis Decumbentis Rhizoma.
引文
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[11]Ki H K,Lee I K,Cheng J P,et al.Benzylisoquinoline alkaloids from the tubers of Corydalis ternate and their cytotoxicity[J].Bioorg Med Chem,2010,20(15):4487-4490.
[12]马宏达,郭涛,颜鸣,等.HPLC法测定夏天无中原阿片碱延胡索乙素和巴马亭[J].解放军药学学报,2009,25(3):208-211.
[2]张铁军,许浚,韩彦琪,等.中药质量标志物(Q-marker)研究:延胡索质量评价及质量标准研究[J].中草药,2016,47(9):1458-1467.
[3]邓敏,宋秀媛,王家富.普洛托品的药理作用研究进展[J].中草药,2001,32(3):275-277.
[4]Li Y,Wang S,Liu Y,et al.Effect of alpha-allocryptopine on transient outward potassium current in rabbit ventricular myocytes[J].Cardiology,2008,111(4):229-236.
[5]郑洪艳.原小檗碱类生物碱作用差异的机理研究[D].天津:天津医科大学,2004.
[6]王新,薛晖,岳媛,等.高表达β1肾上腺素受体细胞膜色谱法靶向筛选夏天无中活性成分[J].中国实验方剂学杂志,2015,21(18):65-68.
[7]张金生,徐任生,陈仲良,等.夏天无化学成分的研究-白毛茛碱宁等的分离和结构[J].中草药,1987,18(l):8.
[8]廖静,梁文藻,涂国士.夏天无中十一个叔胺生物碱的分离和鉴定[J].中国药学英文版,1995,4(2):57-61.
[9]Wang Q,Li Z F,Yang Z H,et al.New alkaloids with anti-inflammatory activities from Corydalis decumbens[J].Phyto Chem Lett,2016,18:83-86.
[10]杨鑫宝,刘扬子,杨秀伟,等.磐安延胡索的化学成分研究[J].中草药,2013,44(16):2200-2207.
[11]Ki H K,Lee I K,Cheng J P,et al.Benzylisoquinoline alkaloids from the tubers of Corydalis ternate and their cytotoxicity[J].Bioorg Med Chem,2010,20(15):4487-4490.
[12]马宏达,郭涛,颜鸣,等.HPLC法测定夏天无中原阿片碱延胡索乙素和巴马亭[J].解放军药学学报,2009,25(3):208-211.