HPLC示差折光法测定水产品中还原糖、磷酸化单糖和蔗糖

Determination of Reducing Sugars,Phosphorylated Sugars and Sucrose in Fish Products by HPLC-RI

DOI：

中文关键词: 水产品 还原糖 磷酸化糖 碱性磷酸酶 HPLC-RI

英文关键词: fish product reducing sugars phosphorylated suagers sucrose HPLC-RI

基金项目:国家“十一五”科技支撑计划项目(2009BAD9B05);浙江省科技计划项目(2008C02015);浙江省重大科技专项(2010C12012)

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中文摘要:

以市售淡水鱼为试材,建立水产品中还原糖(葡萄糖、核糖)、磷酸化糖(磷酸化核糖和磷酸化葡萄糖)和蔗糖的定性、定量检测方法。对鱼肉中各糖进行检测,建立鱼肉中糖的高效液相示差折光分析法检测法。将鱼肉分成背部肌肉和腹部肌肉两个部分,用乙醇/水提取鱼肉中各糖物质,经树脂吸附处理后,对鱼肉中各糖物质进行分析。磷酸化糖检测,首先用碱性磷酸酶脱磷酸化,然后用同法进行检测。结果显示:各糖物质分离效果较好,方法学验证,各物质线性关系良好,回收率在70.2%~98.5%之间,重复性RSD均小于5%。

英文摘要:

The freshwater fish were selected as materials,and a HPLC-RI method for detecting reducing sugars,phosphorylated reducing sugars and sucrose from fish meat and other foods was established in this study.The chromatographic conditions of reducing sugars were investigated and optimized.A Hypersil NH2(4.6 250 mm,2.5 μm) column was used,the mobile phases were 80% ethanol and 20% purified water at the flow rate of 0.6 mL/min,the column temperature was 25 ℃,and the differential refractive index detector temperature was 40 ℃.Ethanol/water(80% v/v) was used to extract mono-and disaccharides from dorsal and belly of fish,and adding resin to remove the amino acids and salts from the extract.The sugars in fish dorsal and belly were determined by the established HPLC-RI method.Phosphorylated sugars were first dephosphorylated by alkaline phosphatase and then determined by the same method.The result showed that the method had a good resolution and good linearity,the recovery rate was between 70.2% and 98.5%,and the RSD of repeatability were all lower than 5%.The result of detection showed that there were differences in category and content of sugars between different parts of fish.

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