柱前衍生高效液相色谱法测定乳与乳制品中羟脯氨酸的含量  

Determination of hydroxyproline in milk and milk products with pre-column derivatization by high performance liquid chromatography

作　　者：魏秀莲[1] 丁美方[1] 董颖超[1] 杨定忠[1] 

机构地区：[1]北京市饲料监察所

出　　处：《中国饲料》2011年第18期41-43,共3页China Feed

摘　　要：本实验建立了羟脯氨酸含量的柱前衍生高效液相色谱法(HPLC)测定乳与乳制品中皮革水解蛋白特征性成分。样品经酸水解后采用异硫氰酸苯酯(PITC)衍生后采用高效液相色谱-紫外可见检测器进行测定。采用AgelaVenusil-AA氨基酸分析专用色谱柱(4.6 mm×150 mm,i.d.5μm),柱温40℃,检测波长254 nm,梯度洗脱,流速1.0mL/min,进样量50μL。方法定量限为0.046μg/mL(S/N=10),高中低不同浓度加标回收率为98.2%～99.0%。该法具有样品处理简单,灵敏度高,回收率高,分析时间短等优点,适用于乳及乳制品中羟脯氨酸含量测定。The experiment established an assay method for hydroxyproline in milk and milk products with pre-column derivatization by high performance liquid chromatograph （HPLC）.After hydrolysis with hydrochloric acid,the samples were derived with phenylisothiocyanate （PITC）.The content of hydroxyproline was determined by （HPLC）and Ultraviolet Detector.The separation of hydroxyproline was performed using a Agela Venusil-AA column （ 150 mm ×4.6 mm i. d. ,5 μm）.The column temperature was kept at 40℃.The mobile phase system was gradient elution with a flow rate at 1.0 mL/min.The detection wavelength was set at 254 nm and the injection volume was 50 μL.The quantification limit was 0.046μg/mL based on a signal-to-noise ratio of 10：1.Recovery rates for the same sample spiked with three levels of hydroxyproline were ranged from 98.2 % to 99.0 %.The method was highly sensitive and selective, pretreatment method was simple and quick,average recovery accorded with the requirement of experiment,and the results was satisfactory.The method can be applied to the detection and analysis of hydroxyproline in milk and milk products.

关 键 词：羟脯氨酸 乳与乳制品 皮革水解蛋白 高效液相色谱法 

分 类 号：S816.17[农业科学—饲料科学]