食品科学 ›› 2022, Vol. 43 ›› Issue (4): 190-197.doi: 10.7506/spkx1002-6630-20201114-159

• 成分分析 • 上一篇    下一篇

超高效液相色谱法测定特殊医学用途婴儿配方食品中的19 种游离氨基酸和牛磺酸

卢兰香,薛霞,宿书芳,张艳侠,尹丽丽,程志,郑文静,孙立臻,王骏,祝建华,刘艳明   

  1. (山东省食品药品检验研究院,山东省特殊医学用途配方食品质量控制工程技术研究中心,山东省食品药品安全检测工程技术研究中心,山东 济南 250101)
  • 出版日期:2022-02-25 发布日期:2022-03-08
  • 基金资助:
    国家食品安全关键技术研发重点专项(2017YEC1601600);山东省重点研发计划(医用食品专项计划)项目(2019YYSP020)

Determination of 19 Free Amino Acids and Taurine in Formulas for Special Medical Purpose Intended for Infants by Ultra-high Performance Liquid Chromatography

LU Lanxiang, XUE Xia, SU Shufang, ZHANG Yanxia, YIN Lili, CHENG Zhi, ZHENG Wenjing, SUN Lizhen, WANG Jun, ZHU Jianhua, LIU Yanming   

  1. (Shandong Institute for Food and Drug Control, Shandong Research Center of Engineering and Technology for Quality Control of Food for Special Medical Purposes, Shandong Research Center of Engineering and Technology for Safety Inspection of Food and Drug, Jinan 250101, China)
  • Online:2022-02-25 Published:2022-03-08

摘要: 建立超高效液相色谱法同时测定特殊医学用途婴儿配方食品中19 种游离氨基酸和牛磺酸的分析方法。样品经α-淀粉酶酶解,调pH值沉淀蛋白等杂质净化,采用6-氨基喹啉基-N-羟基-琥珀酰亚氨基甲酸酯为柱前衍生试剂,以甲酸-甲酸铵缓冲液(20 mmol/L,pH 2.25)为流动相A、甲酸-甲酸铵缓冲液(20 mmol/L,pH 3.00)为流动相B和乙腈为流动相C进行梯度洗脱,经AccQ-Tag Ultra C18色谱柱分离后,二极管阵列检测器检测,外标法定量。结果表明,胱氨酸浓度在5~125 μmol/L之间,其余18 种氨基酸和牛磺酸浓度在10~250 μmol/L范围内线性良好,相关系数均大于0.999;检出限胱氨酸为1.5 μmol/L,其他分析物均为3 μmol/L,定量限胱氨酸为5 μmol/L,其他分析物为10 μmol/L。方法测定的平均回收率为90.16%~109.84%,相对标准偏差均不大于4.77%。该方法操作简单、结果准确、重复性好,可以满足特殊医学用途婴儿配方食品中19 种游离氨基酸和牛磺酸的测定,可为企业质量控制和政府监管提供有力的技术支撑。

关键词: 特殊医学用途婴儿配方食品;氨基酸;牛磺酸;AQC柱前衍生;超高效液相色谱法

Abstract: An ultra-high performance liquid chromatography (UPLC) method was developed for the simultaneous determination of 19 free amino acids and taurine in formulas for special medical purpose intended for infants. The samples were hydrolyzed by α-amylase, and purified by precipitating proteins and other impurities through pH adjustment. The method was based on pre-column derivatization with 6-aminoquinolinyl-N-hydroxysuccinimidate (AQC). Formic acid-ammonium formate (20 mmol/L, pH 2.25), formic acid-ammonium formate (20 mmol/L, pH 3.00), and acetonitrile were used as mobile phases A, B and C for gradient elution, respectively. The analytes were separated on an AccQ-Tag Ultra C18 column, detected with a diode-array detector and quantified by an external standard method. Good linearity was observed for cystine in the range of 5–125 μmol/L and 18 other amino acids and taurine in the range of 10–250 μmol/L with correlation coef?cients higher than 0.999. The limit of detection (LOD) was 1.5 μmol/L for cystine, and 3 μmol/L for the other analytes. The limit of quantitation (LOQ) was 5 μmol/L for cystine, and 10 μmol/L for the other analytes. The average recoveries of the method were in the range of 90.16%–109.84% with relative standard deviation (RSD) not more than 4.77%. The method was easy to operate, accurate and highly reproducible, and could be suitable for the determination of 19 amino acids and taurine in formulas for special medical purpose intended for infants, providing strong technical support for enterprise quality control and government regulation.

Key words: formulas for special medical purpose intended for infants; amino acids; taurine; AQC pre-column derivatization; ultra-high performance liquid chromatography

中图分类号: