示差折光-液相色谱法测定蜂蜜中β-呋喃果糖苷酶残留量  

Determination of β-Fructofuranosidase by Liquid Chromatography with Refractive Index Detector

作　　者：王艳[1] 吴斌[1] 张晓燕[1] 沈崇钰[1] 张睿[1] 李丽花[1] 费晓庆[1] 赵增运[1] 

机构地区：[1]江苏出入境检验检疫局食品实验室,南京210001

出　　处：《分析化学》2012年第10期1602-1606,共5页Chinese Journal of Analytical Chemistry

摘　　要：基于β-呋喃果糖苷酶与棉子糖在柠檬酸-氢氧化钠缓冲介质(pH 4.50)中50℃下反应16 h后的酶解产物蜜二糖含量的变化,建立了高效液相色谱示差折光检测(HPLC-RID)技术测定蜂蜜中β-呋喃果糖苷酶残留量的新方法。以Agilent Carbohydrate柱为分离柱,乙腈-水(78∶22,V/V)为流动相,流速为1.4 mL/min,检测器为示差折光检测器,柱温和检测器温度均为35℃。结果表明,蜂蜜样品经过聚丙烯酰胺凝胶柱分离后,样品峰形得到简化,灵敏度提高了1.3倍;方法的线性范围为10～200 U/kg;测出限为10 U/kg;回收率为81.3%～112.4%;相对标准偏差为3.7%～9.7%(n=6)。当蜂蜜样品中β-呋喃果糖苷酶含量>20 U/kg时,判定为阳性样品,即掺假蜂蜜。A new method for the determination of detecting β-fructofruanosidase in honey was developed by liquid chromatography with refractive index detector（HPLC-RID）.The method was based on the changes of product concentration of melibiose by the reaction of β-fructofruanosidase enzyme with raffinose in the medium of citric acid-sodium hydroxide solution（pH=4.50） for 16 h in water bath at 50 ℃.Agilent Carbohydrate was chosen as the separation column,acetonitrile-water（78 ∶ 22,V/V） was selected as mobile phase with flow rate of 1.4 mL/min.The column temperature and detector temperature was 35 ℃.The results showed that the numbers of peaks were simplified and the sensitivity of this proposed method was improved 1.3 fold as well by using polyacrylamide gel column for pre-separation.A good linear fit was received for the concentration of β-fructofruanosidase enzyme from 10 U/kg to 200 U/kg with the detection limit of 10 U/kg.Recovery was obtained between 81.3% and 112.4% with the relative standard deviation of 3.7% to 9.7%（n=6）.It is judged to be adulteration honey if β-fructofruanosidase concentration is more than 20 U/kg,

关 键 词：Β-呋喃果糖苷酶 蜂蜜 示差折光 液相色谱 掺假 

分 类 号：S896.1[农业科学—特种经济动物饲养] O657.72[农业科学—畜牧兽医]