| DMBA衍生/超高效液相色谱法测定鱼肉中唾液酸含量 |
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| 基金项目:浙江省海洋生物医用制品工程技术研究中心开放基金 |
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| 中文摘要:建立了同时检测鱼肉中N-乙酰神经氨酸（Neu5Ac）、N-羟乙酰神经氨酸（Neu5Gc）和酮基－脱氧壬酮糖酸（KDN）3种核心唾液酸的超高效液相色谱－荧光检测（UPLC－FLD）方法。冷冻干燥后的样品在8 mL 0.6 mol/L盐酸溶液中80 ℃酸解20 min，采用4，5-二甲基-1，2-苯二胺（DMBA）衍生试剂进行衍生。前处理后的样品经Waters Acquity UPLC BEH C18柱（100 mm × 2.1 mm，1.7 μm）分离，以纯水－乙腈为流动相梯度洗脱，荧光检测器检测。结果表明，Neu5Ac、Neu5Gc和KDN在0.02 ~ 5.0 μg/mL范围内呈良好的线性关系（r2 > 0.999），定量下限（S/N = 10）分别为0.20、0.20、0.10 μg/g；加标回收率为90.8% ~ 103%，相对标准偏差（RSD）为2.8% ~ 5.0%。该方法分析时间短，4 min内可完成3种唾液酸的分离，分离效果好，且方法灵敏度、准确度高，适用于鱼肉中唾液酸的测定。 |
| 中文关键词:唾液酸 超高效液相色谱 衍生 鱼肉 |
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| Determination of Sialic Acid in Fish by Ultra High Performance Liquid Chromatography with DMBA Derivatization |
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| Abstract:An ultra high performance liquid chromatography－fluorescence detection（UPLC－FLD） method was developed for the simultaneous determination of 3 core kinds of sialic acids（SAs） including N-acetylneuraminic acid（Neu5Ac），N-glycolylneuraminic acid（Neu5Gc） and 2-keto-3-deoxynononic acid（KDN） in fish. The freeze-dried samples were acid hydrolyzed with 8 mL 0.6 mol/L hydrochloric acid solution for 20 min at 80 ℃，followed by derivatization with 4，5-dimethylbenzene-1，2-diamine（DMBA）．The derivative conditions for DMBA were optimized by single factor test．All the target compounds were separated on an Waters Acquity UPLC BEH C18 column（100 mm × 2.1 mm，1.7 μm） by gradient elution，using pure water and acetonitrile as eluents，then detected by UPLC－FLD．Under the optimized conditions，there were good linear relationships for Neu5Ac，Neu5Gc and KDN in the range of 0.02－5.0 μg/mL（r2 > 0.999），with their limits of quantitation（S/N = 10） of 0.20 μg/g，0.20 μg/g and 0.10 μg/g，respectively．The spiked recoveries for three SAs at three levels ranged from 90.8% to 103%，with relative standard deviations（RSDs） of 2.8%－5.0%．The analysis was rapid，and it could be achieved within 4 min with good separation and sharp peak shapes．With the characteristics of high accuracy and good precision，the method was suitable for the simultaneous determination of Neu5Ac，Neu5Gc and KDN in fish. |
| Key Words:sialic acids ultra performance liquid chromatography derivatization fish |
| 引用本文：李佩佩，严忠雍，陈荫，何鹏飞，黄丽英，方益.DMBA衍生/超高效液相色谱法测定鱼肉中唾液酸含量[J].分析测试学报,2022,41(11):1684-1689. |
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