柱前衍生HPLC法测定开心果中还原型谷胱甘肽(GSH)和总巯基(-SH)

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• 英文论文题名：HPLC Determination of Reduced Glutathione and Total Thiol(-SH) in Pistachio with Precolumn Derivatization
• 论文作者：刘畅 ; 刘亚君 ; 张小勇 ; 崔胜云
• 英文论文作者：Liu Chang ; Liu Ya-jun ; Zhang Xiao-yong ; Cui Sheng-yun ; Key Laboratory of Natural Resources & Functional Molecules of Changbai Mountain of Ministry of Education ; Yanbian University
• 年：2016
• 作者机构：延边大学长白山生物资源与功能分子教育部重点实验室;
• 论文关键词：高效液相色谱法 ; 开心果 ; 谷胱甘肽 ; 巯基化合物
• 会议召开时间：2016-07-01
• 会议录名称：中国化学会第30届学术年会摘要集-第二十三分会：复杂样品分离分析
• 语种：中文
• 分类号：O657.72;TS255.7
• 学会代码：ZGHY
• 会议名称：中国化学会第30届学术年会-第二十三分会 ; 复杂样品分离分析
• 会议地点：中国辽宁大连
• 主办单位：中国化学会
• 学会名称：中国化学会
• 页数：1
• 文件大小：223k
• 原文格式：D
• 会议级别：全国

摘要

用5,5'-二硫双-2-硝基苯甲酸(DTNB)为柱前衍生化试剂,采用细胞破壁、衍生化为一体的同步衍生化提取,HPLC法选择性测定了开心果中GSH和总巯基(-SH)含量。对色谱流动相等条件进行了优化,建立了一种同步测定开心果中含半胱氨酸残基的巯基化合物的测定方法~([1-2])。采用Inertsil ODS-SPC_(18)色谱柱(5μm,4.6 mm×150 mm),流动相为含1%甲酸、0.07%三乙胺的去离子水和乙腈,梯度洗脱,流速为0.8 ml/min,检测波长为327nm。结果表明:开心果中GSH和总巯基(-SH)的含量分别为1.2303μmol/g和4.7718μmol/g,测定方法的回收率达100.52%和101.11%,检测限分别为4.62μmol/L和7.28μmol/L。该方法准确、灵敏度高,为植物细胞中巯基化合物的研究提供了检测手段。
Sample were prepared by simultaneous derivatization during cell rupture.Reduced glutathione(GSH) and total thiols in pistachio were determined by High performance liquid chromatography using 5,5'-dithio-bis-nitrobenzoic acid(DTNB) as derivatization agents.Chromatographic conditions including chromatographic mobile phase are optimized,and established a determination method of simultaneous determination thiol content in the compounds containing cysteine residues~([1-2]).The chromatographic column was Inertsil ODS-SP C_(18)(5μm,4.6 mm×150mm).The elution was performed at a flow rate of 0.8mL/min using the mobile phases of acetonitrile and 1%methanoic acid-0.07%triethylamine deionized water.The detection wavelength was 327 nm.The detection results indicted that abundant GSH and total thiols existed in pistachio with amounts of 1.2303μmol/gμmol/g of GSH and4.7718μmol/g of total thiols.The recoveries of the methods are 100.52%and 101.11%,the detection limits are4.62μmol/L and 7.28μmol/L respectively.The method is sensitive,accurate and suitable for the simultaneous determination of thiol compounds in plant cell.

引文

[1]Donald S.Backos,Christopher C.Franklin,Philip Reigan.Biochemical Pharmacology.2012,83:1005.
    [2]周蓉,曹赵云,牟仁祥,张志刚,柯志成.色谱,2015,30(1):35