反相高效液相色谱法测定酱油中的呈味核苷酸  

Determination of Flavored Nucleotides in Soy Sauce by RP-HPLC Method

作　　者：黄文武 冯纬 黄光荣 HUANG Wen-wu;FENG Wei;HUANG Guang-rong(Huzhou Institute for Food and Drug Control, Huzhou 313000, China;Hangzhou FoodBrewing Co., Ltd., Hangzhou 310021, China;College of Life Sciences,China Jiliang University, Hangzhou 310018, China)

机构地区：[1]湖州市食品药品检验研究院,浙江湖州313000 [2]杭州市食品酿造有限公司,杭州310021 [3]中国计量大学生命科学学院,杭州310018

出　　处：《中国调味品》2019年第1期150-153,共4页China Condiment

基　　金：杭州市科技计划项目(20150432B28);湖州市科技计划项目(2016GY06)

摘　　要：文章建立了酱油中呈味核苷酸(鸟苷酸GMP和肌苷酸IMP)含量测定的反相高效液相色谱方法。该方法中样品以甲醇脱蛋白、C18固相萃取小柱脱色和除杂质,以50mmol/L pH 4.6的磷酸缓冲液为流动相进行等梯度洗脱,流速为1.0mL/min,检测波长为254nm。结果显示:GMP和IMP的保留时间分别为12.2,13.8min,区分度很好,酱油样品在25min内可完全分离,GMP和IMP在5.00～50.00mg/L范围内具有良好的线性,相关系数R2均可达0.9999,变异系数分别为2.08%和1.83%,方法检出限(RSN=3)分别为0.52,0.23mg/L,定量限(RSN=10)分别为1.74,0.77mg/L,酱油样品的平均加标回收率在90.0%～105.0%之间。该方法快速准确、灵敏度高、重复性好,适用于酱油中呈味核苷酸的测定。In this paper, a revised-phase high-performance liquid chromatography (RP-HPLC) method for the determination of flavored nucleotides, including inosine- 5'-monophosphate (IMP)and guanosine-5'-monophosphate (GMP) in soy sauce is established. In this method, the sample is deproteined by methanol and decolored, decontaminated with a C 18 solid-phase extraction column. And then the sample is determined by RP-HPLC with 1.0 mL/min 50 mmol/L pH 4.6 phosphate buffer at 254 nm. The result shows that GMP and IMP are isolated and the retention time is 12.2, 13.8 min respectively. And the total determination could be finished within 25 min. The GMP and IMP content is linear correlated with the peak area at 5.00~50.00 mg/L , with the linear correlation coefficient (R 2 ) of 0.9999. The coefficients of variation for GMP and IMP determination are 2.08% and 1.83% respectively, and the limits of detection (RSN=3) and the limits of quantitation (RSN=10) are 0.52, 0.23 mg/L and 1.74, 0.77 mg/L respectively. The recovery rates are 90.0%~105.0% when the established method is used for the determination of IMP and GMP in soy sauce. This method is rapid, accurate, has high sensitivity and good repeatability, and it could be used for the determination of flavored nucleotides in soy sauce.

关 键 词：呈味核苷酸 反相高效液相色谱 酱油 鸟苷酸 肌苷酸 

分 类 号：TS264.21[轻工技术与工程—发酵工程]