高通量检测大样本桑黄发酵液总黄酮含量方法的建立  

High-throughput Quantitative Assay of Total Flavonoids in Large-scale Samples

作　　者：毛东霞 阮芹芹 赵敏[2] 郭丹丹[1] 王苗[1] 田晓雪[1] 刘陶[1] 

机构地区：[1]陕西师范大学生命科学学院,西安710100 [2]长庆油田分公司油气工艺研究院,西安710021

出　　处：《天然产物研究与开发》2015年第10期1706-1711,1788,共7页Natural Product Research and Development

基　　金：陕西省特种资源开发与利用项目(693102);中央高校科研业务费特别支持项目(1301030208)

摘　　要：建立了大样本的桑黄发酵液样品中总黄酮的高通量测定方法。通过L9（3^4）正交实验设计优化，确定了基于亚硝酸钠．硝酸铝比色法的微孔板检测方法，优化后总黄酮含量测定的最佳反应条件：5％亚硝酸钠30μL，10％硝酸铝30μL，10％氢氧化钠50μL，室温反应10min，检测波长为510nm。检测结果表明，针对大样本检测，该法稳定性高，平均回收率为100．7％，相对标准偏差为0．607％，线性范围为0—50μg／mL，线性相关系数r^2=0．9994。比较分析结果显示，和多种传统总黄酮测定方法相比，实验所建立的高通量测定方法检测大样本快速、准确、用药量小，且稳定性高。The aim of this study was to develop a high-throughput quantitative assay for detecting the total flavonoids in large-scale samples from the broth of Phelllnus sp.. The orthogonal experimental design was preformed according to L9 （34 ） orthogonal table using 96 microplate. Based on the variance analysis of the results from the orthogonal experiments, the optimum experimental ehromogenie conditions were obtained as follows： the addition volume of 5% sodium nitrite, 10% aluminum nitrate,10% sodium hydroxide were 30 μL,30 μL,100 μL,respeetively,and then the sample should be kept for 10 rain at room temperature. The maximum absorption wavelength for detecting the total flavonoids from the broth ofPhellinus sp. was determined to be 510 nim. The results showed that the developed method was precision and stable. The average recovery was 100.7%. The relative standard deviations of the evaluation test was 0. 607%, and the linear range was 0-50 μg/mL （r2 = 0.9994）. The comparative results between different methods indicated that the developed high-throughput assay for the total flavonoids content determination assay of large-scale samples hereby was more accurate, rapid, reliable and using less sample than the traditional method.

关 键 词：高通量 超微量微孔板分光光度计 亚硝酸钠-硝酸铝比色法 桑黄 总黄酮 

分 类 号：Q93[生物学—微生物学] TS3[轻工技术与工程]