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| Determination of Sialic Acid in Fish by Ultra High Performance Liquid Chromatography with DMBA Derivatization |
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| DOI： |
| KeyWord:sialic acids ultra performance liquid chromatography derivatization fish |
| Author | Institution |
| LI Pei-pei，YAN Zhong-yong，CHEN Yin，HE Peng-fei，HUANG Li-ying，FANG Yi |
1. Marine Fisheries Research Institute of Zhejiang，Zhoushan ，China;
2. Key Lab of Sustainable Utilization of Technology Research for Fishery Resource of Zhejiang Province，Zhoushan ，China;
3. Marine Biomedical Products Engineering Technology Research Center of Zhejiang Province，Zhoushan ，China |
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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. |
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