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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| Published in | 东北农业大学学报 Vol. 48; no. 5; pp. 58 - 64 |
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| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
东北农业大学食品学院,哈尔滨,150030
2017
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| Subjects | |
| Online Access | Get full text |
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| Abstract | 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。 |
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| AbstractList | 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。 TS252.7; 利用酸水解释放出的糖基化产物中氨基葡萄糖,采用Elite C18色谱柱分离,以68％的0.025 mol· L-1pH 3.6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1 mL· min-1,进样体积10 μL,柱温35℃.定量分析结果表明,氨基葡萄糖0.1～100 μg·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、1.0 mg 3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0.02 μg·mL-1.糖基化乳清蛋白中氨基葡萄糖含量为2.03 mg· g-1.RP-HPLC测定法用于乳清蛋白-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. |
| Abstract_FL | 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 x 250 mm) utilizing an isocratic elution of acetic acid sodium acetate buffer solution (containing 68％ 0.025 mol· L-1 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 μL 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 0.02 μg·mL-1.Content of glucosamine in glycosylated whey protein was 2.03 mg·g1.Therefore,this method had the characteristics of simple operation,high reproducibility and sensitivity,It could be widely applied to determine glucosamine in glycosylated protein. |
| Author | 张英华 刘艳乐 刘天舒 朱敏 |
| AuthorAffiliation | 东北农业大学食品学院,哈尔滨150030 |
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| Author_FL | LIU Yanle LIU Tianshu ZHANG Yinghua ZHU Min |
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| DocumentTitleAlternate | Analysis of glucosamine in glycosylated whey protein-glucosamine using reversed-phase high-performance liquid chromatography |
| DocumentTitle_FL | Analysis of glucosamine in glycosylated whey protein-glucosamine using reversed-phase high-performance liquid chromatography |
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| Keywords | 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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| Notes | 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 |
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| Snippet | ... TS252.7; 利用酸水解释放出的糖基化产物中氨基葡萄糖,采用Elite C18色谱柱分离,以68％的0.025 mol· L-1pH 3.6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1 mL· min-1,进样体积10 μL,柱温35℃.定量分析结果表明,氨基葡萄糖0.1～100... |
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| SubjectTerms | 乳清蛋白 反相高效液相色谱 氨基葡萄糖 |
| Title | RP-HPLC测定乳清蛋白糖基化产物中的氨基葡萄糖 |
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