摘要
目的:建立柱前衍生-反相高效液相色潽法测定乳及乳制品中L-羟脯氨酸含量。方法:乳及乳制品中蛋白质经酸水解后,用2,4二硝基氟苯对L-羟脯氨酸进行衍生,采用250 mm×4.6 mm,5μm Dia-monsilAAA分析柱进行分离,检测波长360 nm,流动相为0.02 mol/L磷酸氢二钠与磷酸二氢盐缓冲液、乙腈、甲醇,流速1.0 ml/mim,采用梯度洗脱。结果:L-羟脯氨酸的浓度与峰面积成良好的线性关系,相关系数r为0.9994,回收率在94.0%～97.4%之间,样品的测定结果良好。结论:本方法测定乳及乳制品中L-羟脯氨酸灵敏度较高,干扰因素少,结果稳定、准确,适合乳及乳制品中L-羟脯氨酸的测定。
Objective：Precolumn derivatization-Reversed phase high performance liquid chromatography method（RP-HPLC） was established for determination of L-Hydroxyproline in dairy and dairy products.Methods： Samples were hydrolyzed in HCl.2,4-Dinitrofluorobenzene,a derivatization reagent,was used for sample derivatization.A 5 μm DiamonsilAAA column was used for separation with a mobile phase consist of 0.02 mol/L NaH2PO4/ Na2HPO4-methanol-Acentrile（9∶ 1） for gradient elution.The detection wavelength was at 360 nm.Results： The results showed that concentration of L-Hydroxyproline and peak areas achieved a nice linear relation,r was 0.9994.The recovery was from 94.0% to 97.4%.Conclusion： The proposed method was stable,accurate,high sensitive,less-interference and suitable for determination of L-Hydroxyproline in dairy and dairy products.
出处
《中国卫生检验杂志》
北大核心
2012年第9期2056-2057,2060,共3页
Chinese Journal of Health Laboratory Technology
关键词
高效液相色谱法
柱前衍生
L-羟脯氨酸
乳及乳制品
Reversed Phase High performance liquid chromatography
Precolumn derivatization
L-Hydroxyproline
Dairy and dairy products
