Citation: WANG Yan, WU Bin, ZHANG Xiao-Yan, SHEN Chong-Yu, ZHANG Rui, LI Li-Hua, FEI Xiao-Qing, ZHAO Zeng-Yun. Determination of β-Fructofuranosidase by Liquid Chromatography with Refractive Index Detector[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(10): 1602-1606. doi: 10.3724/SP.J.1096.2012.20111 shu

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

1.
Laboratory of Food, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing 210001, China

Corresponding author: WANG Yan,
Received Date: 10 February 2012
Available Online: 18 April 2012

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,
- β-Fructofruanosidase,
- Honey,
- Refractive index detector,
- Liquid chromatography,
- Adulteration
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沈阳化工大学材料科学与工程学院沈阳 110142