Optimization of extraction technology and determination of sodium hyaluronate in solid drinks by ethanol precipitation combined with enzymatic hydrolysis- reversed phase chromatography
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(1. Institute of Analysis, Guangdong Academy of Sciences 〔China National Analytical Center, Guangzhou〕, Guangzhou, Guangdong 510070, China; 2. Guangdong Provincial Key Laboratory of Chemical Measurement and Emergency Test Technology, Guangzhou, Guangdong 510070, China; 3. Guangdong Provincial Engineering Research Center for Efficacy Component Testing and Risk Substance Rapid Screening of Health Food, Guangzhou, Guangdong 510070, China)

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    Abstract:

    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 C18 (4.6 mm×250 mm,5 μm) column with mobile phase: acetonitrile 10 mmol/L sodium octane sulfonate solution (Vacetonitrile∶Voctane 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 0.999 9. 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.

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张志舟,林嘉婷,潘灿盛,等.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[J].食品与机械英文版,2023,39(5):64-69.

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  • Received:October 03,2022
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  • Online: June 09,2023
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