半夏白术天麻汤物质基准HPLC指纹图谱及多成分定量分析

Fingerprint and multicomponent quantitative analysis of Banxia Baizhu Tianma Decoction by HPLC

目的建立半夏白术天麻汤的HPLC指纹图谱,并同时测定7种成分(天麻素、甘草苷、柚皮苷、橙皮苷、异甘草苷、白术内酯Ⅲ、甘草次酸)的含量,旨在为半夏白术天麻汤的质量控制提供依据。方法采用Inert Sustain C18色谱柱(250 mm×4.6 mm,5μm)对其进行分离,采用乙腈-水溶液为流动相进行梯度洗脱,流速为1.0 mL/min,检测波长254 nm,柱温30℃。建立18批半夏白术天麻汤样品的HPLC指纹图谱,并进行相似度评价,对共有峰进行归属和指认,测定指认出的成分含量。结果18批样品指纹图谱与对照指纹图谱的相似度均>0.983;共标定25个共有峰,其中2个来自半夏,1个来自天麻,6个来自白术,6个来自橘红,2个来自茯苓,5个来自甘草,另外3个共有峰不能明确其来源。通过对共有峰进行指认,发现8号峰为天麻素,14号峰为甘草苷、15号峰为柚皮苷,16号峰为橙皮苷,19号峰为异甘草苷,21号峰为白术内酯Ⅲ及25号峰为甘草次酸。7种成分的含量测定结果分别为天麻素2.1933~5.0626 mg/g、甘草苷0.1220~0.2867 mg/g、柚皮苷7.3165~15.8486 mg/g、橙皮苷0.1301~0.3928 mg/g、异甘草苷0.1733~0.4524 mg/g、白术内酯Ⅲ0.2361~0.7849 mg/g、甘草次酸0.0180~0.0372 mg/g。结论所建立的半夏白术天麻汤HPLC指纹图谱及定量测定方法,稳定性、重复性均较好,可为半夏白术天麻汤质量控制和评价提供参考。

Objective To establish HPLC fingerprint of Banxia Baizhu Tianma Decoction,and simultaneously determine the contents of seven components(gastrodin,liquiritin,naringin,hesperidin,isoliquiritin,atractylodes lactoneⅢ,glycyrrhetinic acid).To provide the basis for the quality control of Banxia Baizhu Tianma Decoction.Methods The separation was carried out on an Inert Sustain C18 column(250 mm×4.6 mm,5μm).The mobile phase was acetonitrile-water solution,gradient elution was used.Volume flow rate was 1.0 mL/min.Detection wavelength was 254 nm.Column temperature was 30℃.HPLC fingerprints of 18 batches of Banxia Baizhu Tianma Decoction samples were established,the similarity was evaluated.The common peaks were assigned and identified,and the quantity of components identified was determined.Results The similarity of fingerprints of the control and 18 batches of samples were greater than 0.983.And 25 common peaks were calibrated,including 2 from Pinellia Ternata,1 from Gastrodia Elata,6 from Atractylodes Macrocephala,6 from Exocarpium Citri Rubrum,2 from Poria Cocos and 5 from Glycyrrhiza Uralensis Fisch,of which 2 common peaks could not be clearly identified.Peak 8 was gastrodin.Peak 14 was liquiritin.Peak 15 was naringin.Peak 16 was hesperidin.Peak 19 was isoliquiritin.Peak 21 was atractylodes lactoneⅢ.Peak 25 was glycyrrhetinic acid.Contents of the seven components respectively were:gastrodin 2.1933-5.0626 mg/g,liquiritin 0.1220-0.2867 mg/g,naringin 7.3165-15.8486 mg/g,hesperidin 0.1301-0.3928 mg/g,isoliquiritin 0.1733-0.4524 mg/g,atractylodes lactoneⅢ0.2361-0.7849 mg/g,glycyrrhetinic acid 0.0180-0.0372 mg/g.Conclusion HPLC fingerprint and quantitative determination method of Banxia Baizhu Tianma Decoction have merits of good repeatability and stability,which provide the reference for quality control and evaluation.