乙醇沉淀结合酶解—高效液相色谱法测定固体饮料中透明质酸钠  

Optimization of extraction technology and determination of sodium hyaluronate in solid drinks by ethanol precipitation combined with enzymatic hydrolysis-reversed phase chromatography

作　　者：张志舟 林嘉婷 潘灿盛 张飞 胡含含 ZHANG Zhi-zhou;LIN Jia-ting;PAN Can-sheng;ZHANG Fei;HU Han-han(Institute of Analysis,Guangdong Academy of Sciences〔China National Analytical Center,Guangzhou〕,Guangzhou,Guangdong 510070,China;Guangdong Provincial Key Laboratory of Chemical Measurement and Emergency Test Technology,Guangzhou,Guangdong 510070,China;Guangdong Provincial Engineering Research Center for Efficacy Component Testing and Risk Substance Rapid Screening of Health Food,Guangzhou,Guangdong 510070,China)

机构地区：[1]广东省科学院测试分析研究所〔中国广州分析测试中心〕,广东广州510070 [2]广东省化学测量与应急检测技术重点实验室,广东广州510070 [3]广东省保健食品功效成分检测与风险物质快速筛查工程技术研究中心,广东广州510070

出　　处：《食品与机械》2023年第5期64-69,共6页Food and Machinery

基　　金：广东省科学院测试分析研究所(中国广州分析测试中心)自筹科研资金项目(编号:202103)。

摘　　要：目的:建立反相液相色谱测定固体饮料中透明质酸钠的分析方法。方法:以透明质酸钠含量为评价指标,样品提取液经乙醇沉淀后,采用透明质酸酶对沉淀物进行酶解处理,通过单因素试验和正交试验法优化得到最佳酶解条件:酶解时间120 min,酶解温度34℃,pH值6.5。色谱柱为Ulitimate C18(4.6 mm×250 mm, 5μm),流动相:乙腈10 mmol/L辛烷磺酸钠溶液(V乙腈∶V辛烷磺酸钠=10∶90),流速0.6 mL/min;检测波长232 nm,外标法定量。结果:透明质酸钠的相关系数为0.999 9,加标回收率96.4%~101%,相对标准偏差3.16%,检出限为0.062 g/kg。结论:乙醇沉淀结合酶解—反相色谱专属性强、准确度高、重复性好和干扰较小,适用于固体饮料中透明质酸钠的含量测定。Objective:This study aimed to establish the methods for determining the content of sodium hyaluronate in solid drinks by reversed-phase high performance liquid chromatography.Methods:With sodium hyaluronate content as evaluation index,after the sample extract was precipitated by ethanol,hyaluronase was used to hydrolyze the precipitate.The optimal enzymolysis conditions obtained by single factor test and orthogonal test were as follows:enzymolysis,at 34℃and pH 6.5,for 120 min.The determination was performed on Uliumate C 18(4.6 mm×250 mm,5μm)column with mobile phase:acetonitrile 10 mmol/L sodium octane sulfonate solution(V acetonitrile∶V octane sulfonate=10∶90)at flow rate of 0.6 mL/min.The detection wavelength was 232 nm and quantified by external standard method.Results:The correlation coefficient of sodium hyaluronate was 0.9999.The recoveries of samples were between 96.1%~101%,and the relative standard deviation was 3.16%.The determination of limit was 0.062 g/kg.Conclusion:Ethanol precipitation combined with enzymolysis-reversed phase chromatography has strong specificity,high accuracy,good repeatability and less interference,which is suitable for the determination of sodium hyaluronate in solid drinks.

关 键 词：固体饮料 透明质酸钠 反相液相色谱法 乙醇沉淀 酶解 

分 类 号：TS278[轻工技术与工程—农产品加工及贮藏工程] O657.72[轻工技术与工程—食品科学与工程]