RP-HPLC测定乳清蛋白糖基化产物中的氨基葡萄糖

利用酸水解释放出的糖基化产物中氨基葡萄糖，采用EliteC18色谱柱分离，以68％的O．025mol·L^-1pH3．6乙酸-乙酸钠缓冲溶液和32％甲醇作为流动相等度洗脱，流速为1mL·min^-1，进样体积10灿，柱温35℃。定量分析结果表明，氨基葡萄糖0．1～100μg·mL^-1与氨基葡萄糖峰面积线性关系良好（R2=0．9998）；以0．3、0．5、1．0mg3个添加水平作回收试验，氨基葡萄糖平均回收率为88．05％～110．79％，相对标准偏差为2．45％~5．92％；氨基葡萄糖检出限为0．02μg·m^-1。糖基化乳清蛋白中氨基葡萄糖含量为2．03mg·g^-1。RP-HPLC测定法...

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Bibliographic Details
Published in东北农业大学学报 Vol. 48; no. 5; pp. 58 - 64
Main Author 张英华 刘艳乐 刘天舒 朱敏
Format Journal Article
LanguageChinese
Published 东北农业大学食品学院,哈尔滨,150030 2017
Subjects
乳清蛋白
反相高效液相色谱
氨基葡萄糖
glucosamine
whey protein
乳清蛋白
氨基葡萄糖
反相高效液相色谱
reversed-phase high-performance liquid chromatography (RP-HPLC)
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Summary:利用酸水解释放出的糖基化产物中氨基葡萄糖，采用EliteC18色谱柱分离，以68％的O．025mol·L^-1pH3．6乙酸-乙酸钠缓冲溶液和32％甲醇作为流动相等度洗脱，流速为1mL·min^-1，进样体积10灿，柱温35℃。定量分析结果表明，氨基葡萄糖0．1～100μg·mL^-1与氨基葡萄糖峰面积线性关系良好（R2=0．9998）；以0．3、0．5、1．0mg3个添加水平作回收试验，氨基葡萄糖平均回收率为88．05％～110．79％，相对标准偏差为2．45％~5．92％；氨基葡萄糖检出限为0．02μg·m^-1。糖基化乳清蛋白中氨基葡萄糖含量为2．03mg·g^-1。RP-HPLC测定法用于乳清蛋白一氨基葡萄糖糖基化产物制备条件优化，方法简单、重复性好、灵敏度高，可用于蛋白质-氨基葡萄糖糖基化产物中氨基葡萄糖含量分析测定。
Bibliography:reversed-phase high-performance liquid chromatography （RP-HPLC）; whey protein;glucosamine
ZHANG Yinghua, LIU Yanle, LIU Tianshu, ZHU Min（School of Food Sciences, Northeast Agricultural University, Harbin 150030, China）
23-1391/S
Glucosamine in glycosylated whey protein was released hydrochloric acid hydrolysis. The RP-HPLC separation was performed on an Elite C18 （Hypersil ODS25 μm 4.6 mm ×250 mm） utilizing an isocratic elution of acetic acid sodium acetate buffer solution （containing 68% 0.025 mol. L1 pH 3.6） and methyl alcohol （containing 32%） as the mobile phase at a flow rated of 1 mL. min-1. Injiction volume and column temperature were set at 10 IJL and 35 ℃. The results indicate that the linear ranage was from 0.1 to 100 μg·mL^-1 for glucosamine and correlation coefficient （R2 was greater than 0.99）. The average recoveries spiked at the three concentration levels of 0.30, 0.50, 1.00 mg ranaged between 88.05% and 110.79% with the relative standard deviations from 1.22% to 5.92%. The limit of detection was 0
ISSN:1005-9369