摘要
该研究建立了一种亲水性高效液相色谱同时测定婴幼儿配方乳粉中胞嘧啶核苷酸、尿嘧啶核苷酸、次黄嘌呤核苷酸、鸟嘌呤核苷酸和腺嘌呤核苷酸共5种核苷酸含量的检测方法。婴幼儿配方乳粉样品经超纯水提取其中的核苷酸,沉淀蛋白后上清液用Oasis PRiME HLB固相萃取柱净化,采用Waters BEH Amide色谱柱分离,以0.1%磷酸溶液和乙腈为流动相进行梯度洗脱,二极管阵列检测器采集信号,外标法定量。结果表明,5种核苷酸在2~40 mg/L内线性关系良好(R^(2)>0.999),方法的检出限和定量限分别为0.5~1.0 mg/kg和1.5~3.0 mg/kg;空白样品添加6、15、30 mg/kg三个浓度水平,平均加标回收率为90.17%~101.20%,相对标准偏差为1.6%~4.5%(n=6)。该方法快速准确、基质干扰小、灵敏度高,适用于婴幼儿配方乳粉中核苷酸的测定。
A method for simultaneous determination of five nucleotides(cytidine monophosphate,uridine monophosphate,inosine monophosphate,guanosine monophosphate,and adenosine monophosphate)in infant formula by hydrophilic high performance liquid chromatography was established.The nucleotides in infant formula samples were extracted with ultrapure water and then purified with Oasis PRiME HLB solid-phase extraction column after being deproteinated,separated on a Waters BEH Amide column with a mobile phase consisting of 0.1%(V/V)phosphoric acid-water and acetonitrile by gradient elution,finally determined using diode array detector and quantitated with external standard method.Five nucleotides showed good linearity in the concentration range of 2-40 mg/L with the correlation coefficients greater than 0.999,the limits of detection(LOD)and limits of quantitation(LOQ)were 0.5-1.0 mg/kg and 1.5-3.0 mg/kg,respectively.The average recoveries of blank samples spiked at 6 mg/kg,15 mg/kg and 30 mg/kg were 90.17%-101.20%,with relative standard deviation(RSD)values of 1.6%-4.5%(n=6).This method proved to be quick,accurate,less matrix interference,highly sensitive and suitable for the simultaneous determination of five nucleotides in infant formula.
作者
黄坤
张菊
王会霞
HUANG Kun;ZHANG Ju;WANG Huixia(Hubei Provincial Institute for Food Supervision and Test,Key Laboratory of Detection Technology of Focus Chemical Hazards in Animal-derived Food for State Market Regulation,Hubei Provincial Engineering and Technology Research Center for Food Quality and Safety Test,Wuhan 430075,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2024年第10期312-317,共6页
Food and Fermentation Industries
基金
国家市场监督管理总局科技计划项目(2022MK087)。
关键词
核苷酸
亲水性高效液相色谱
婴幼儿配方乳粉
通过式固相萃取净化
nucleotides
hydrophilic high performance liquid chromatography
infant formula
pass-through solid phase extraction purification
作者简介
第一作者:黄坤,硕士,高级工程师;通信作者:王会霞,正高级工程师,E-mail:972567848@qq.com。
