柱前衍生化HPLC法测定植物水解蛋白中的17种氨基酸  

Determination of 17 Amino Acids of HVP by Pre-Column Derivatization HPLC

作　　者：陈建芳 江学文[1] 梁科 王莹莹 CHEN Jianfang;JIANG Xuewen;LIANG Ke;WANG Yingying(College of Life Science,South China Normal University,Guangzhou 510631,China;Guangdong Dayuan Oasis Food Safety Technology Co.,Ltd.,Guangzhou 510631,China)

机构地区：[1]华南师范大学生命科学学院,广东广州510631 [2]广东达元绿洲食品安全股份有限公司,广东广州510631

出　　处：《食品安全导刊》2023年第11期99-104,共6页China Food Safety Magazine

摘　　要：本文建立了一种2,4-二硝基氟苯柱前衍生测定植物水解蛋白(Hydrolyzed Vegetable Protein,HVP)中17种氨基酸的高效液相色谱方法。色谱柱为Diamonsil AAA,柱温为45℃,检测波长为360nm,进样体积为10μL,流速为1.0 m L·min^(-1);流动相A为0.02 mol·L^(-1)Na(2)HPO_(4)+0.02mol·L^(-1)NaH_(2)PO_(4)水溶液,流动相B为甲醇∶乙腈=10∶90(V∶V),梯度洗脱,17种氨基酸得到了很好的分离,衍生方法操作简单,方法精密度高、稳定性和重复性均良好,适用于植物水解蛋白中氨基酸的定量测定。HVP与酿造酱油中的氨基酸组成差异较大,主要体现在谷氨酸、天冬氨酸、精氨酸、甘氨酸以及脯氨酸,可为酿造酱油与配制酱油的鉴别提供一定依据。This article establishes a high-performance liquid chromatography method for the determination of 17 amino acids in hydrolyzed vegetable protein(HVP)using pre column derivatization of 2,4-dinitrofluorobenzene.The chromatographic column is DiamonsilAAA,with a column temperature of 45℃,detection wavelength of 360 nm,and injection volume of 10μL.The flow rate was 1.0 mL·min^(-1);the mobile phase A was a 0.02 mol·L^(-1)Na_(2)HPO_(4)+0.02 mol·L^(-1)NaH_(2)PO_(4)aqueous solution,and the mobile phase B was methanol:acetonitrile=10∶90(V∶V).Gradient elution was performed,and 17 amino acids were well separated.The derivation method was simple,with high precision,stability,and repeatability,making it suitable for the quantitative determination of amino acids in plant hydrolyzed proteins.The amino acid composition of HVP and fermented soy sauce is quite different,mainly reflected in glutamic acid,aspartic acid,arginine,glycine and proline,which can provide some basis for the identification of fermented soy sauce and prepared soy sauce.

关 键 词：植物水解蛋白(HVP) 氨基酸 2 4-二硝基氟苯 高效液相色谱法 酿造酱油 

分 类 号：O657.72[理学—分析化学] TS264.21[理学—化学]