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高效液相色谱法测定蜂蜜中β-呋喃果糖苷酶酶活
引用本文:王介平,郭军,陈杨,王海燕,李睿,王永亮.高效液相色谱法测定蜂蜜中β-呋喃果糖苷酶酶活[J].蜜蜂杂志,2011,31(12):21-23.
作者姓名:王介平  郭军  陈杨  王海燕  李睿  王永亮
作者单位:重庆市畜牧科学院,重庆荣昌,402460
摘    要:建立了一种检测蜂蜜中β-呋喃果糖苷酶酶活的方法,即样液中加棉籽糖作为反应底物,在pH为5.4的缓冲液中,45aC摇床200r/min条件下反应2h,测定样液中的密二糖含量。色谱条件为:Phenomenex NH2 Luan 5U 250×4.60mm;流动相:乙腈:水=80:20;进样量:10μL;流速:1.2mL/min;柱温:40℃;示差折光检测器,温度40cc。实验结果表明:β-呋喃果糖苷酶酶活在0.3—1.7U/mL范围内线性良好,相关系数R为0.998 10。相对标准偏差为1.72%(n=5),平均回收率为94.5%。该分析方法简便、准确、灵敏,重现性好,适用于蜂蜜中β-呋喃果糖苷酶酶活的检测。

关 键 词:掺假蜂蜜  液相色谱法(HPLC)  β-呋喃果糖苷酶  棉籽糖

Assay of Beta-fructofuranosidase Activity in Honey by High Cerformance Liquid Chromatography
WANG Jie-ping,GUO Jun,CHEN Yang,WANG Hai-yang,LI Rui,WANG Yong-liang.Assay of Beta-fructofuranosidase Activity in Honey by High Cerformance Liquid Chromatography[J].Journal of Bee,2011,31(12):21-23.
Authors:WANG Jie-ping  GUO Jun  CHEN Yang  WANG Hai-yang  LI Rui  WANG Yong-liang
Institution:WANG Jie-ping,GUO Jun,CHEN Yang,WANG Hai-yang,LI Rui,WANG Yong-liang(ChongQing Academy of Animal Sciences,Rongchang,Chongqing 402460,China)
Abstract:We analyzed the concentration of melibiose in the sample solution,which reacted 2h after add raffinose as reaction substrate (reaction conditions: pH 5.4 buffer solution,45℃ shaking table 200r/min).At last,A method was developed for the determination of beta-fructofuranosidase activity in honey. The sample was separated with NH2 column (250 ×4.60mm, 5u) at 40℃, which was then eluted with 80% acetonitrile aqueous solution at a rate of 1.2mL/min,and the signal was detected by RID.The results showed that the linear range for beta-fructofuranosidase activity were in the range of 0.3-1.7 u/mL with a correlation coeficient of 0.998 10. The relative standrd deviations for beta-fructo- furanosidase activity were 1.72% (n=5) and the average recoveries were 94.5%.This method is simple, accurate, sensi- tive and better reproducibility.
Keywords:honey adulterated  high performance liquid chromatography  beta-fructofuranosidase  raffinose  
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