贵州医科大学学报

2016, v.41;No.193(10) 1176-1180

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贵州不同地区山银花中4种异绿原酸含量测定方法的建立
Establish of HPLC Method for Determination of Four Isochlorogenic Acids of Flos Lonicerae Produced in Different Regions in Guizhou

印酬;向红霞;王洪凤;孟小夏;张敏;高秀丽;
YIN Chou;XIANG Hongxia;WANG Hongfeng;MENG Xiaoxia;ZHANG Min;GAO Xiuli;College of pharmacy,Guizhou Medical University;

摘要(Abstract):

目的:建立测定贵州不同地区山银花中4种异绿原酸含量的高效液相色谱(HPLC)方法。方法:考察HPLC法[色谱柱为Diamonsil-C18(250 mm×4.6 mm,5μm)、流动相为乙腈(A)-0.4%磷酸(B),梯度洗脱、流速为1m L/min,柱温30℃,检测波长327 nm]测定山银花中新绿原酸、绿原酸、异绿原酸A(3,5-二咖啡酰奎宁酸)、异绿原酸C(4,5-二咖啡酰奎宁酸)含量的色谱条件、线性关系及回收率,观察贵州不同产地的10批山银花中的4种异绿原酸的含量。结果:本研究设定的色谱条件可测得山银花中4种异绿原酸的色谱峰,山银花中新绿原酸、绿原酸、异绿原酸A、异绿原酸C分别在2.73~87.45 mg/L(r=0.999 9)、14.24~455.95 mg/L(r=0.999 8)、5.75~184.0 mg/L(r=0.999 3)及5.25~168.0 mg/L(r=0.999 7)范围内呈良好线性关系,山银花中的4种异绿原酸回收率均>99.2%;10批山银花中新绿原酸的含量以遵义最高,绿原酸、异绿原酸A及异绿原酸C含量最高的采集地均为毕节,毕节地区山银花中4种异绿原酸的总含量也高于其他地区。结论:成功建立了测定山银花异绿原酸的含量的HPLC法,贵州山银花中的4种异绿原酸含量不相同。
Objective: To establish an HPLC method for simultaneous determination of four isochlorogenic acids of Flos Lonicerae produced in different regions in Gui Zhou. Methods: The chromatographic separation was carried out on a Diamonsil-C18column( 250 mm × 4. 6 mm,5 um) with gradient elution of acetonitrile and 0. 4% phosphoric acid at a flow rate of 1m L / min. Column temperature 30 ℃and the detection wave-length was 327 nm. The chromatographic conditions,linear relationship and recovery rate were determined in detecting the content of neochlorogenic acid,chlorogenic acid,isochlorogenic acid A( 3,5-two caffeoylquinic acid,isochlorogenic acid C( 4,5-two caffeoylquinic acid).The contents of 4 different chlorogenic acid were evaluated and compared between 10 batch of Flos Lonicerae produced in different area of Guizhou province. Results: 4 different chromatographic peaks of chlorogenic acid in Flos Lonicerae on the chromatographic conditions set in this study can be measured. The chromatographic peak of neochlorogenic acid,chlorogenic acid,isochlorogenic acid A and isochlorogenic acid C was in 2. 73 ~ 87. 45 mg / L( r = 0. 999 9),14. 24 ~ 455. 95 mg / L( r = 0. 999 8),5. 75 ~ 184. 0 mg / L( r = 0. 999 3) and 5. 25 ~ 168. 0( mg / L r = 0. 999 7),respectively. A good linear relationship was showed within these ranges of 4 different chlorogenic acid in Flos Lonicerae and recovery rate was over 99. 2%. In the 10 batch of Flos Lonicerae,the content of neochlorogenic acid was the highest in Flos Lonicerae produced in Zunyi. The content of chlorogenic acid,isochlorogenic acid A and isochlorogenic acid C were the highest in Flos Lonicerae produced in Bijie. The total content of4 kinds of isochlorogenic acid in Flos Lonicerae produced in Bijie area were higher than those of Flos Lonicerae produced in other areas. Conclution: The HPLC method for the determination of the content of chlorogenic acid in Flos Lonicerae is successfully established. The content of 4 kinds of different chlorogenic acid in Flos Lonicerae produced in different areas of Guizhou is not the same.

关键词(KeyWords): 山银花;有机酸;色谱法,高压液相;贵州
Flos Lonicerae;organic acids;chromatography;high pressure liquid;Guizhou

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技成果转化引导基金计划项目[黔科合成转字(2015)5202]

作者(Author): 印酬;向红霞;王洪凤;孟小夏;张敏;高秀丽;
YIN Chou;XIANG Hongxia;WANG Hongfeng;MENG Xiaoxia;ZHANG Min;GAO Xiuli;College of pharmacy,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2016.10.013

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