高效液相色谱法检测酱油中羟脯氨酸的含量  

Detection of hydroxyproline in soy sauces by high performance liquid chromatography

作　　者：胡华[1,2] 吕定 杜青 张惟广[2] 朱永红 

机构地区：[1]重庆市计量质量检测研究院国家农副加工产品及调味品质量监督检验中心,重庆400020 [2]西南大学食品科学学院,重庆400716

出　　处：《中国调味品》2010年第4期106-109,共4页China Condiment

基　　金：重庆市科技攻关计划项目(CSTC;2007AC1021);国家质检公益性行业科研专项项目(10-100)

摘　　要：实验建立了柱前衍生高效液相色谱分析酱油中羟脯氨酸(hydroxyproline,Hyp)含量的方法。样品经乙醇提取,以2,4-二硝基氟苯为衍生试剂,采用GeminiC18(4.6×250mm,5μm)色谱柱分离,检测波长为360nm。Hyp质量浓度与其峰面积在10～320mg/L范围内具有良好的线性关系,相关系数为0.9992。回收率为97.19%～110.18%,检出限为2.5mg/L。对实际样品中相对Hyp含量(表示为每克氨态氮中所含羟脯氨酸的毫克数)进行检测发现,纯酿造酱油在0.67～1.93mg/g之间,酸水解植物蛋白液在9.51～20.11mg/g之间,酸水解猪皮和猪肌肉分别为855.55,37.03mg/g,三类样品间存在较明显的差异。上述结果提示,Hyp并非胶原蛋白中所特有,但通过检测酱油样品中Hyp含量有可能区分酱油中是否添加酸水解蛋白液。An analytical method of hydroxyproline（Hyp） in soy sauces was established with pre-column derivatization by high performance liquid chromatography. The samples were extracted with ethanol and then derivatized with 2,4-dinitrofluorobenzene. Chromatographic separation was performed with Gemini C18 column （4. 6×250 mm, 5 μm）, and detection wavelength was 360 mm. There was good linear correlation between the concentrations and peak area of Hyp in the range of 10 - 320mg/L, the correlation coefficient was 0. 9992. The recovery was 97. 19%-110. 18%, and the limit of detection was 2. 5 mg/L. The relative amounts of Hyp （which expressed as milligrams of Hyp per gram of ammonia nitrogen） were determined in different samples. The relative amounts of Hyp were apparently different in samples detected. It ranged from 0. 67 to 1.93 mg/g for fermented soy sauces and from 9.51 to 20.11 mg/g for acid hydrolyzed vegetable protein solutions. The relative amounts of Hyp in acid hydrolyzed pigskins and pork were 855. 55 and 37. 03 mg/g respectively. The results indicated that the collagen protein was not the sole source of Hyp, but it was possible that we could identify whether the samples adulterated with acid hydrolyzed protein by detecting Hyp in soy sauces.

关 键 词：羟脯氨酸 酱油 水解动物蛋白 高效液相色谱 检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]