科研之友
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摘要
A new method for the determination of detecting beta-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 beta-fructofruanosidase enzyme with raffinose in the medium of citric acid-sodium hydroxide solution (pH=4.50) for 16 h in water bath at 50 degrees C. 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 degrees C. 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 beta-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 beta-fructofruanosidase concentration is more than 20 U/kg,
- 出版日期2012-10
