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桑椹多酚HPLC指纹图谱的研究

HPLC Fingerprint of Phenolic Compounds in Mulberry

【作者】 孙伟

【导师】 王华;

【作者基本信息】 西北农林科技大学 , 发酵工程, 2007, 硕士

【摘要】 桑椹含有多种酚类物质,包括矢车菊素、天竺葵素、碧冬茄素、白藜芦醇、芦丁、槲皮素、金丝桃苷、绿原酸、桑色素等,具有很好的保健与医疗作用,这些物质的种类和含量受基因和环境条件的影响,从而会影响桑椹及其产品的质量,建立桑椹多酚的HPLC指纹图谱对控制桑椹及其产品的质量非常重要。实验通过超声波辅助提取法,优化了桑椹多酚的最佳提取工艺;通过试凑试验法,建立了桑椹多酚的HPLC检测条件;并利用该HPLC检测条件,建立了桑椹多酚的HPLC指纹图谱。研究结果表明:(1)桑椹多酚的最佳提取工艺是:最佳提取剂是pH为2.0的纯甲醇,提取温度为25℃,提取时间为20min,料液比为1:12,超声波频率为60kHz,提取次数为4次。(2)桑椹多酚HPLC指纹图谱检测条件为:色谱柱:Shimpack VP-ODS(250mm×4.6mm,5μm);流动相A:含0.05%磷酸的水,流动相B:乙腈:四氢呋喃=50:50(v/v);梯度洗脱程序:0-15-28-34-34-0min、0-8-28-40-40-0B%、100-92-72-60-60-100A%;流速:0.8mL/min;检测波长为313nm;柱温为30℃;进样量为20μL。(3)桑椹多酚HPLC指纹图谱本试验所建立的大十和红果2号桑椹多酚HPLC指纹图谱,其样品收集、制备及色谱条件优选均符合HPLC中药指纹图谱的技术要求和建立原则。其方法学考察结果和指纹图谱评价结果,均符合国家中药指纹图谱的标准,说明本实验建立的桑椹HPLC指纹图谱共有模式可以用于建立不同地区的桑椹HPLC指纹图谱,并且可以作为科学评价桑椹及其产品质量的检测方法。

【Abstract】 Mulberry has good efficacy in health protection and medical care mostly due to its rich polyphenols, such as cyanidin, pelargonidin, chlorogenic acid, gallic acid, resveratrol, rutin, quercetin, morin, ursolic acid andβ-sitosterol, which are powerfull antioxidants. These polyphenols are greatly responsible for the quality of mulberry and phenolic compound fingerprint can give a full evaluation of mulberry quality, so it is necessary to build mulberry fingerprint for its quality assessment.In this study, we optimized the extraction technology of mulberry polyphenolics using ultrasonic extraction, built a suitable HPLC condition for mulberry polyphenolics detection by Cut-and-Try Procedure, and finally built the chromatography fingerprints of mulberry polyphenolics by HPLC.The main results were as follows:1. The best conditions for extraction technique of mulberry polyphenols are: Ultrasonic extraction at 25℃under 60Hz, 100%MeOH as extractant, the suitable pH 2.0, extracted time 20min, the ratio of mulberry pomace and extraction 1:12 and four-time extraction.2. The suitable conditions for HPLC dectection of mulberry polyphenols are: A column of Shimpack VP-ODS (250mm×4.6mm, 5μm) was employed. Solvent A: water/phosphoric acid = 99.95/0.05 (v/v); Solvent B: ACN/TDF = 50/50 (v/v); the flow rate, 0.8 mL/min; a linear gradient profile consisting of solvent A with the following proportions (v/v) of solvent B: 0min, 0%B; 0-15min, 0-8%B; 15-28min, 8-28%B; 28-34min, 28-40%B; 34-45min, 40-0%B; the detecting wavelength, 313nm; the temperature, 30℃; the inject volume, 20μL.3. HPLC fingerprints of polyphenols in Dashi and Hongguo 2 were obtained: In this research, the collection and the preparation of samples, the confirmation of the HPLC condition, the result of methodology survey, the consequences of fingerprint assessment, all accord with the national standards in technology and principles of fingerprint establishment, so the method for establishing the chromatography fingerprints of mulberry polyphenolics by HPLC in this study, can be able to a common mode to build the mulberry fingerprints of different varieties and different areas, and also can be a reliable means for appraisal and assessment the quality of mulberry and the products made from mulberry.

【关键词】 桑椹多酚HPLC指纹图谱
【Key words】 MulberrypolyphenolicsHPLC Fingerprint
  • 【分类号】R284
  • 【被引频次】13
  • 【下载频次】1152
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