高效液相色谱-荧光检测法测定红肉及其加工肉中唾液酸含量  

Determination of Sialic Acid in Red Meat and Its Processed Products by High Performance Liquid Chromatography-Fluorescence Detection

作　　者：赵非 陈宝英 李克峰 王旭 ZHAO Fei;CHEN Baoying;LI Kefeng;WANG Xu(State Key Laboratory of Food Nutrition and Safety,Tianjin University of Science&Technology,Tianjin 300457,China;Tianjin SunnyPeak Biotech Co.Ltd.,Tianjin 300457,China)

机构地区：[1]天津科技大学,省部共建食品营养与安全国家重点实验室,天津300457 [2]天津桑尼匹克生物科技有限公司,天津300457

出　　处：《食品科学》2021年第4期313-318,共6页Food Science

基　　金：天津市自然科学基金项目(16JCQNJC1440);天津科技大学省部共建食品营养与安全国家重点实验室开放课题(SKLFNS-KF-201820)。

摘　　要：采用高效液相色谱-荧光检测法对红肉及其加工肉中两种唾液酸N-乙酰神经氨酸(N-acetylneuraminic acid,Neu5Ac)和N-羟乙酰神经氨酸(N-glycolylneuraminic acid,Neu5Gc)进行定性和定量分析。利用单因素试验对衍生化与样品酸解条件进行优化,并借助超声缩短酸解时间。结果表明,以盐酸作为酸解试剂,超声辅助酸解30 min,4,5-亚甲二氧基-1,2-邻苯二胺盐作为衍生化试剂,衍生化试剂浓度为13 mmol/L,样品与衍生化试剂比值为1∶1,衍生化时间120 min时,检测效果最佳。Neu5Ac和Neu5Gc在0.1~10μg/mL范围内线性良好,回收率为91.2%~119.7%,检出限分别为0.003 mg/kg和0.01 mg/kg,重复性的相对标准偏差(relative standard deviation,RSD)为0.7%~1.8%,精密度RSD分别为1.4%和1.2%。本方法具有灵敏度高、分析时间短、重复性及准确性好等特点。A high performance liquid chromatography-fluorescence detection(HPLC-FLD)method was developed for the qualitative and quantitative analysis of the sialic acids N-acetylneuraminic acid(Neu5Ac)and N-glycolylneuraminic acid(Neu5Gc)in red meat and its processed products.Single factor experiments were carried out to optimize the derivatization and acid hydrolysis conditions.Besides,ultrasound-assisted acidification was used to shorten the acidification time.It was found that the optimal sample preparation procedure involved ultrasound-assisted acidification time for 30 min with hydrochloric acid as the hydrolysis reagent and derivatization for 120 min with the same volume of 13 mmol/L 4,5-methylenedioxy-1,2-phenylenediamine dihydrochloride(DMB)as the derivatization reagent.The calibration curves for Neu5Ac and Neu5Gc displayed good linearity in the range of 0.1–10μg/mL.The recoveries were in the range of 91.2%–119.7%.The limits of detection(LODs)for Neu5Ac and Neu5Gc were 0.003 and 0.01 mg/kg,respectively.The precision expressed as relative standard deviations(RSDs)for the two sialic acids was in the range of 0.7%–1.8%and the repeatability RSDs were 1.4%and 1.2%,respectively.The HPLC-FLD method has the characteristics of high sensitivity,short analysis time,and good repeatability and accuracy.

关 键 词：高效液相色谱-荧光检测 红肉 加工肉 N-乙酰神经氨酸 N-羟乙酰神经氨酸 

分 类 号：TS207.3[轻工技术与工程—食品科学]