微孔板法与传统方法对比测定花菇、猴头菇和茶树菇的抗氧化活性  

Comparison on the determination results of microplate method and traditional spectrophotometric method on antioxidant activity of Flower Mushroom,Hericium and Agrocybe Aegerita

作　　者：王明娟[1] 武彦舒[2] 李竹[3] 王琳琳[3] 王晨琳[3] 

机构地区：[1]河南中医学院第三附属医院第一药剂科,郑州450008 [2]陕西省榆林市第二医院药剂科,陕西榆林719000 [3]河南中医学院药学院,郑州450046

出　　处：《中国当代医药》2015年第16期7-11,共5页China Modern Medicine

基　　金：国家自然科学基金资助项目(81473435);河南中医学院科研苗圃工程项目(MP2011-06);河南中医学院大学生创新学习项目(YXCX〔2014〕22);河南中医学院研究生创新基金项目(2014YCX009)

摘　　要：目的对比微孔板法和传统方法分光光度法测定花菇、猴头菇和茶树菇体外抗氧化活性的差异。方法超声波法提取3种菇的水提物和醇提物,以VC为对照品,均与底物1,1-二苯基-2-三硝基苯肼自由基(DPPH·)反应,其中微孔板法为微孔板结合酶标仪于517 nm波长处快速测定菇类提取物的DPPH·抑制率,传统分光光度法于紫外-可见分光光度计517 nm波长处检测。对2种方法的测定结果进行曲线拟合,并进行方法重复性对比。结果 2种方法抗氧化结果 IC50基本一致,曲线拟合度较好,差异无统计学意义(P>0.05);其中对照品VC的微孔板法拟合模型公式为y=23.872Lnx+152.526,分光光度法模型公式为y=22.469Lnx+146.044,2种方法的模型中系数和常数经t检验差异有统计学意义(P<0.01),提示重叠率高。分别对3种样品的提取液进行曲线拟合,结果显示3种菇提取液2种抗氧化活性测定结果的曲线拟合度均较好,差异有统计学意义(P<0.05)。实验中微孔板法试剂用量较小,精密度更高。结论 2种菇类体外抗氧化活性测定方法实验结果一致,差异不明显,微孔板法在试剂用量和精密度方面略具优势。Objective To compare the determination results about microplate method and traditional spectrophotometric method on the antioxidant activity in vitro of Flower Mushroom,Hericium and Agrocybe Aegerita. Methods Ultrasonic extraction was used to extract the aqueous extracts and ethanol extracts of 3 mushrooms,VC was used as the reference substance,and given the reaction with the substrate 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-（2,4,6-trinitrophenyl） hydrazyl free radical （DPPH·）.Microplate method was that using microplate combined with microplate reader in 517 nm wavelength for determination of DPPH·inhibition ratio of mushroom extracts fastly,traditional spec-trophotometric method was that determining in the UV Vis spectrophotometry in 517 nm wavelength for determination. Then the curve fitting and method repeated comparison was given,and method repeatability comparison was done. Re-sults Two methods of antioxidant results IC50 were consistent relatively,curve fitting was better,and no statistical differ-ence （P〉0.05）; microplate method fitting model formula of reference substance VC was y=23.872Lnx＋152.526,spec-trophotometry methed was y=22.469Lnx＋146.044,the model of the two methods’ coefficients and constants was signifi-cant （P〈0.01） by the t-test,with high overlapping rate.Then respectively curve fitting on extracts of three kinds samples, the results showed that the determination results of two method curve fitting were good of three mushrooms extracts’.

关 键 词：抗氧化活性 微孔板法 分光光度法 

分 类 号：R965[医药卫生—药理学]