摘要
目的建立HPLC法测定银杏叶提取物中9种小分子有机酸类成分莽草酸、儿茶素、表儿茶素、没食子酸、原儿茶酸、6-羟基犬尿喹啉酸、对羟基苯甲酸、对羟基肉桂酸、咖啡酸含量的方法,并结合多元统计分析方法比较不同厂家生产的银杏叶提取物间质量差异。方法采用Inertsil ODS-3 C18(250 mm×4.6 mm,5μm)色谱柱;以乙腈-0.4%磷酸水溶液为流动相,梯度洗脱,波长切换测定(220 nm检测莽草酸、儿茶素、表儿茶素,254 nm检测没食子酸、原儿茶酸、6-羟基犬尿喹啉酸、对羟基苯甲酸,310 nm检测对羟基肉桂酸、咖啡酸),柱温40℃。结果不同厂家生产的银杏叶提取物中有机酸总含量有差异,厂家内部样品中各有机酸含量同样存在差异。经聚类分析、主成分分析、相关与回归分析等多元统计方法分析,筛选出儿茶素、没食子酸、原儿茶酸及6-羟基犬尿喹啉酸为体现样品质量差异的特征成分,同时成分间具有内在相关性。结论建立的方法操作简便、重复性好、结果可靠,可用于银杏叶提取物中有机酸类成分的质量评价。筛选出的儿茶素、没食子酸、原儿茶酸及6-羟基犬尿喹啉酸是体现质量差异的特征成分,又是具有内在关联的共性成分,为提升银杏叶提取物整体质量控制能力提供了依据,同时表明银杏叶提取物的提取工艺并不统一,企业内部工艺的稳定性也有待提高。
Objective A method for simultaneous determination of nine active organic acids [shikimic acid, catechinic,(-)-epicatechin, gallic acid, protocatechuic acid, 6-hydroxykynurenic acid(6 HKA), p-hydroxybenzoic acid(PHBA), hydroxycinnamic acid, and caffeic acid] in Ginkgo biloba extracts(GBEs) by HPLC wavelength switching method was established, and its quality was evaluated by statistical analysis. Methods An Inertsil ODS-3 C18 column(250 mm×4.6 mm, 5 μm) was used with a column temperature of 40 ℃ and a mobile phase gradient of acetonitrile-0.4% phosphoric acid. The detection wavelengths were 220 nm [shikimic acid, catechinic,(-)-epicatechin], 254 nm(gallic acid, protocatechuic acid, 6 HKA, PHBA), 310 nm(hydroxycinnamic acid, caffeic acid), respectively. Results Remarkable differences were found among the nine compositions in different producing areas samples. The samples from different manufacturers showed significant difference in organic acid compounds. There were great difference of the contents of organic acid in various batches from different manufacturers. The data were processed through unsupervised principal component analysis(PCA), cluster analysis(CA), correlation, and regression analysis to compare the quality differences. Catechinic, gallic acid, protocatechuic acid, and 6 HKA were recognized as characteristic chemical markers that contributed most to reflect the difference between samples, and there were internal relations among these compounds. Conclusion The established method was simple, accurate, and reliable with good precision, repeatability and stability, which can be used for the simultaneous determination of nine organic acid compounds. Through multivariate statistical analysis, the Q-markers that may affect the difference of GBEs were better identified, which can lay the foundation for the comprehensive quality control of GBEs. Meanwhile, the extracting process of GBEs were not uniform. The stability of process parameter in the enterprise was prepared to be further enhanced.
作者
赵一懿
殷海利
郭洪祝
傅欣彤
陈有根
ZHAO Yi-yi;YIN Hai-li;GUO Hong-zhu;FU Xin-tong;CHEN You-gen(Beijing Institute for Drug Control,NMPA Key Laboratory for Quality Evaluation of Traditional Chinese Medicine(Traditional Chinese Patent Medicine),Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine,Beijing 102206,China;College of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)
出处
《中草药》
CAS
CSCD
北大核心
2020年第14期3679-3685,共7页
Chinese Traditional and Herbal Drugs
基金
国家中医药管理局中药标准化资助项目(ZYBZH-C-HEB-16)。
关键词
银杏叶提取物
有机酸
多元统计分析
儿茶素
没食子酸
原儿茶酸
6-羟基犬尿喹啉酸
莽草酸
表儿茶素
对羟基苯甲酸
对羟基肉桂酸
咖啡酸
Ginkgo biloba extracts
organic acid
multivariate statistical analysis
catechinic
gallic acid
protocatechuic acid
6-hydroxykynurenic acid
shikimic acid
(-)-epicatechin
p-hydroxybenzoic acid
hydroxycinnamic acid
caffeic acid
