食品科学 ›› 2021, Vol. 42 ›› Issue (4): 313-318.doi: 10.7506/spkx1002-6630-20191008-031

• 安全检测 • 上一篇    下一篇

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

赵非,陈宝英,李克峰,王旭   

  1. (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)
  • 出版日期:2021-02-25 发布日期:2021-02-25
  • 基金资助:
    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)

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

ZHAO Fei, CHEN Baoying, LI Kefeng, WANG Xu   

  1. (1. State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology, Tianjin 300457, China; 2. Tianjin SunnyPeak Biotech Co. Ltd., Tianjin 300457, China)
  • Online:2021-02-25 Published:2021-02-25

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0.1~10 μg/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。

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

Abstract: 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 0.1–10 μg/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.

Key words: high performance liquid chromatography-fluorescence detection; red meat; processed meat products; N-acetylneuraminic acid; N-glycolylneuraminic acid

中图分类号: