文章摘要
仝爽,陈建武,王成友,杜浩,危起伟.长江鲟科鱼类尿液成分中L-色氨酸的检测分析[J].上海海洋大学学报,2019,28(4):550-558
长江鲟科鱼类尿液成分中L-色氨酸的检测分析
Determination and analysis of L-tryptophan in urine component of Acipenseriformes in Yangtze River
投稿时间:2018-07-20  修订日期:2018-09-28
DOI:10.12024/jsou.20180702367
中文关键词: 中华鲟  长江鲟  L-色氨酸  高效液相色谱  尿液
英文关键词: Acipenser sinensis  Acipenser dabryanus  L-tryptophan  high performance liquid chromatography (HPLC)  urine
基金项目:国家自然科学基金青年科学基金(31602160)
作者单位E-mail
仝爽 上海海洋大学 海洋科学学院, 上海 201306
中国水产科学研究院长江水产研究所 农业农村部淡水生物多样性保护重点实验室, 湖北 武汉 430223 
 
陈建武 农业农村部淡水鱼类种质监督检验测试中心, 湖北 武汉 430223  
王成友 中国水产科学研究院长江水产研究所 农业农村部淡水生物多样性保护重点实验室, 湖北 武汉 430223 wangcy@yfi.ac.cn 
杜浩 中国水产科学研究院长江水产研究所 农业农村部淡水生物多样性保护重点实验室, 湖北 武汉 430223  
危起伟 中国水产科学研究院长江水产研究所 农业农村部淡水生物多样性保护重点实验室, 湖北 武汉 430223  
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中文摘要:
      采用高效液相色谱-荧光法(HPLC-FLD)建立了中华鲟和长江鲟尿液中L-色氨酸的测定方法。中华鲟和长江鲟尿液样品由V(甲醇)∶V(乙腈)=1∶1提取,离心后,上清液经ENVI-18固相萃取小柱净化,利用symmetry@C18色谱柱(250 mm×4.6 mm,5 μm)进行分离,以乙腈-1%(体积分数)甲酸水溶液(10∶90,体积分数)为流动相等度洗脱,在流速1.0 mL/min,荧光波长λex=285 nm,λem=353 nm条件下进行检测。结果显示:L-色氨酸标准品的峰面积与其质量浓度在0.010~2.000 μg/mL范围内线性关系良好,线性回归方程为y=247.27x-0.125 5(R2=1)。L-色氨酸的检出限(LOD)为0.005 μg/mL,定量限(LOQ)为0.020 μg/mL。L-色氨酸空白加标回收率为82.2%~102.1%,样品基质加标回收率为85.6%~95.1%,相对标准偏差(RSD)均小于5.7%。本方法有良好的回收率和精密度,且步骤简便,经济省时,可用于中华鲟和长江鲟尿液L-色氨酸的检测,为研究L-色氨酸在濒危鲟鱼中华鲟和长江鲟信息交流方面的作用奠定方法学基础。
英文摘要:
      A method of high performance liquid chromatography-fluorescence spectroscopy (HPLC-FLD) was used to determinate L-tryptophan in urine of Acipenser sinensis and A. dabryanus. The samples were extracted by methanol and acetonitrile, by centrifugation to get supernatant, and purified by an ENVI-18 solid-phase extraction column. The acetonitrile-1% (volume fraction) formic acid aqueous solution (10:90, volume fraction) was used as the mobile phases, the extracts were separated on a symmetry@C18 chromatographic column (250 mm×4.6 mm, 5 μm). The samples were determinated under the condition of the flow rate of 1.0 mL/min, with the fluorescence wavelength of λex=285 nm, λem=353 nm. The results indicated that the significant linear relationships between peak areas and mass concentrations of the analytes were obtained in the range of 0.010-2.000 μg/mL (R2=1), and linear regression equation was y=247.27x-0.125 5. The limits of detection (LOD) and limits of quantification (LOQ) of L-tryptophan were 0.005 μg/mL and 0.020 μg/mL. The recoveries of blank matrix and sample substrate with standard samples were 82.2%-102.1% and 85.6%-95.1%, respectively, with less than 5.7% of the relative standard deviations (RSD). This method has some obvious advantage such as easy operation, time saving, high recoveries and high accuracies. It will be valuable to analyze L-tryptophan in urine of A. sinensis and A. dabryanus. A methodology basis is provided to study the role of L-tryptophan in information exchange of A. sinensis and A. dabryanus.
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