柴胡龙骨牡蛎汤高效液相色谱指纹图谱的建立
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摘要
目的:建立柴胡龙骨牡蛎汤水煎液和正丁醇萃取液的反相高效液相指纹图谱的检测方法。方法:采用高效液相色谱法,Thermo Fisher Hypersil GOLD C18色谱柱,流动相乙腈-0.1%磷酸水溶液,梯度洗脱,柱温35℃,流速1 m L·min-1,检测波长276,210 nm。结果:分别建立了柴胡龙骨牡蛎汤水煎液和正丁醇萃取液指纹图谱共有模式,并指认其中的共有色谱峰。通过与对照品的保留时间和紫外光谱图比较,水煎液指纹图谱中8,12,14,15号峰分别归属为黄芩苷、肉桂酸、桂皮醛、大黄酸。正丁醇萃取液指纹图谱中16,17,28,29号峰分别归属为人参皂苷Rg1,人参皂苷Re,柴胡皂苷a,柴胡皂苷b2。10批柴胡龙骨牡蛎汤水煎液和正丁醇萃取液的HPLC指纹图谱相似度,均>0.88。结论:该方法灵敏度高、稳定性和准确性良好,为柴胡龙骨牡蛎汤质量标准建立提供了可靠的科学依据。
Objective: To establish the chromatographic fingerprint for Chaihu Longgu Muli Tang and nbutanol extraction by RP-HPLC-DAD. Method: HPLC method was applied to establish the chromatographic fingerprint. The separation was performed on a Thermo Fisher Hypersil GOLD C18 column with a gradient elution composed of acetonitrile and 0. 1% phosphoric acid. The column temperature was 35 ℃ and the flow rate was 1. 0m L·min- 1. The detective wavelength was 276,210 nm. Result: The common pattern of chromatographic fingerprint of Chaihu Longgu Muli Tang and n-butanol extraction was established, and 4 peaks of them were identified respectively. In decoction,the 8th,12 th,14th and 15 th peaks were assigned to baicalin,cinnamic acid,cinnamaldehyde and rhein respectively by comparison with the retention times and UV spectra of reference substances. The 16 th, 17 th, 28 th and 29 th peaks were assigned to ginsenoside Rg1, ginsenoside Re,saikosaponin a and saikosaponin b2 respectively of n-butanol extraction. The similarity of 10 batches of Chaihu Longgu Muli Tang and n-butanol extraction were all more than 0. 88. Conclusion: The established HPLC fingerprint of Chaihu Longgu Muli Tang has desirable precision, reproducibility, and can provide a basis for quality evaluation of Chaihu Longgu Muli Tang.
Objective: To establish the chromatographic fingerprint for Chaihu Longgu Muli Tang and nbutanol extraction by RP-HPLC-DAD. Method: HPLC method was applied to establish the chromatographic fingerprint. The separation was performed on a Thermo Fisher Hypersil GOLD C18 column with a gradient elution composed of acetonitrile and 0. 1% phosphoric acid. The column temperature was 35 ℃ and the flow rate was 1. 0m L·min- 1. The detective wavelength was 276,210 nm. Result: The common pattern of chromatographic fingerprint of Chaihu Longgu Muli Tang and n-butanol extraction was established, and 4 peaks of them were identified respectively. In decoction,the 8th,12 th,14th and 15 th peaks were assigned to baicalin,cinnamic acid,cinnamaldehyde and rhein respectively by comparison with the retention times and UV spectra of reference substances. The 16 th, 17 th, 28 th and 29 th peaks were assigned to ginsenoside Rg1, ginsenoside Re,saikosaponin a and saikosaponin b2 respectively of n-butanol extraction. The similarity of 10 batches of Chaihu Longgu Muli Tang and n-butanol extraction were all more than 0. 88. Conclusion: The established HPLC fingerprint of Chaihu Longgu Muli Tang has desirable precision, reproducibility, and can provide a basis for quality evaluation of Chaihu Longgu Muli Tang.
引文
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[2]邓暖繁.柴胡龙骨牡蛎汤治疗恶性肿瘤化疗后抑郁症临床观察[J].光明中医,2012,27(1):76-78.
[3]尚俊平.柴胡加龙骨牡蛎汤治疗老年抑郁症30例[J].甘肃中医,2010,23(2):247-248.
[4]王洪然.柴胡加龙骨牡蛎汤治疗中风后抑郁临床观察[J].中国实用医药,2011,6(7):175-176.
[5]石月平,赵建宇,周亚滨.柴胡加龙骨牡蛎汤治疗心血管神经症体会[J].中医药信息,2006,19(3):53.
[6]蔡华,叶方,黄良永,等.超高效液相质谱联用法研究柴胡特征指纹图谱[J].中国医院药学杂志,2014,34(5):359-364.
[7]赵恒强,王晓,周洁,等.山东产区黄芩HPLC定量指纹图谱研究[J].中国现代中药,2014,16(7):528-533.
[8]王举涛,徐文方,刘金旗,等.HPLC-ELSD法测定黄芪中黄芪甲苷的含量[J].安徽医药,2006,10(1):28-29.
[9]刘金欣,孟繁蕴,魏英勤,等.UPLC-ELSD同时测定北柴胡中柴胡皂苷a,c,d的含量[J].中国实验方剂学杂志,2013,19(5):83-86.
[10]刘晓帆,娄子洋,朱臻宇,等.采用HPLC-TOF/MS对中药复方小柴胡汤中化学成分的快速分析鉴别[J].第二军医大学学报,2009,30(8):941-946.
[11]林佳,徐丽珍,刘江云.中药桂枝的HPLC指纹图谱研究[J].中成药,2006,28(2):169-171.