食品科学 ›› 2018, Vol. 39 ›› Issue (12): 174-178.doi: 10.7506/spkx1002-6630-201812027

• 生物工程 • 上一篇    下一篇

高效液相色谱法评价海洋生物中半胱氨酸双加氧酶活性

范子瑞,胡佳妃,江瑞妮,杨星玲,金火喜*   

  1. (浙江海洋大学食品与医药学院,浙江?舟山 316022)
  • 出版日期:2018-06-25 发布日期:2018-06-15
  • 基金资助:
    浙江海洋大学人才引进科研启动项目(Q1420); 2016年浙江省大学生科技创新活动计划暨新苗人才计划项目(2016R411039); 2016年舟山市科技局浙江海洋大学专项(2016C41002)

Evaluation of Cysteine Dioxygenase Activity in Marine Organisms by High Performance Liquid Chromatography

FAN Zirui, HU Jiafei, JIANG Ruini, YANG Xingling, JIN Huoxi*   

  1. (School of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, China)
  • Online:2018-06-25 Published:2018-06-15

摘要: 建立半胱亚磺酸的高效液相色谱分析方法,并利用该方法测定不同海洋生物组织中半胱氨酸双加氧酶的活性。应用邻苯二甲醛(o-phthalaldehyde,OPA)为衍生试剂进行柱前衍生,色谱柱为Agilent Eclipse XDB-C18。结果发现,当检测波长为315?nm、半胱亚磺酸与OPA物质的量比1∶2、衍生时间3?min、流动相甲醇-乙酸钠(8∶2,V/V)、流速1?mL/min时,半胱亚磺酸在10~400?μg/mL质量浓度范围内具有良好的线性关系(R2>0.99),平行样品的相对标准偏差为0.9%~6.4%,保留时间为1.46?min。以此方法为基础,对不同海产品组织中半胱氨酸双加氧酶活性进行检测,结果显示,所选择的海洋贝类、蟹类、虾类、鱼类中均检测到半胱氨酸双加氧酶活性,其中南美白对虾中活性最高。不同组织比较,鱼体肝脏中半胱氨酸双加氧酶活性最高,而鳃中活性最低。

关键词: 海产品, 半胱氨酸双加氧酶, 半胱亚磺酸, 高效液相色谱

Abstract: An analytical method for cysteine sulfonate by high performance liquid chromatography (HPLC) was established and applied to determine the activity of cysteine dioxygenase in different marine organism tissues. Cysteine sulfinate was subjected to 3 min pre-column derivatization with o-phthalaldehyde (OPA) at a molar ratio of 1:2. The separation was performed on an Agilent Eclipse XDB-C18 using a mobile phase composed of methanol and sodium acetate (8:2, V/V) at a flow rate of 1 mL/min. The analyte was detected at 315 nm. The calibration curve showed a good linearity (R2 > 0.99) in the concentration range of 10–400 μg/mL. The relative standard deviations (RSDs) for parallel samples were 0.9%–6.4% and the retention time was 1.46 min. Using this method, cysteine dioxygenase activity was detected to be present in marine shellfish, crab, shrimp and fish, with the highest activity being observed in Penaeus vannamei. Among different tissues of fish, the highest activity of cysteine dioxygenase was detected in the liver, while the lowest was found in the gill.

Key words: seafood, cysteine dioxygenase, cysteine sulfinate, high performance liquid chromatography (HPLC)

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