柱后衍生高效液相色谱法检测膝沟藻毒素

食品科学 ›› 2013, Vol. 34 ›› Issue (2): 170-174.

柱后衍生高效液相色谱法检测膝沟藻毒素

姜沛宏,章超桦,秦小明,卢虹玉,林华娟,梁颖瑜

广东海洋大学食品科技学院

收稿日期:2011-11-12 修回日期:2012-12-20 出版日期:2013-01-25 发布日期:2013-01-15
通讯作者: 林华娟 E-mail:huajuanlin@vip.163.com

Analysis of gonyautoxins (GTXs) by a post-column derivatization high performance liquid chromatography

Received:2011-11-12 Revised:2012-12-20 Online:2013-01-25 Published:2013-01-15
Contact: Hua-Juan LIN E-mail:huajuanlin@vip.163.com

摘要/Abstract

摘要：

本研究探讨柱后衍生条件和流动相条件对膝沟藻毒素群(GTXs，麻痹性贝毒的一类)的反应荧光强度和分离效果的影响。柱后衍生条件分析结果显示，各GTXs对反应液pH值、温度、甲酰胺体积分数以及甲酸铵浓度等柱后衍生反应条件的感度有较大差异。确定最佳柱后衍生反应条件为70mmol/L高碘酸与含0.12mol/L氢氧化钾和0.75mol/L甲酸铵的20%甲酰胺溶液的混合液作为柱后衍生反应液、pH7.3～7.5、流速0.8min/mL、柱后反应温度50℃。流动相条件分析结果显示，甲醇体积分数、庚烷磺酸钠浓度、pH值和磷酸浓度均显著影响各GTXs的保留时间和分离效果，确定最佳流动相条件为甲醇体积分数1%、庚烷磺酸钠浓度1.5mmol/L、磷酸浓度10mmol/L、pH7.3。灵敏度分析结果显示，除GTX2无明显差异外，GTX1、GTX3和GTX4的灵敏度分别高于大岛法4.7、3.4和3.8倍。

Abstract:

Paralytic shellfish poisoning (PSP) toxins is an important risk for sea food safety, and detecting technology of high performance liquid chromatography (HPLC) for PSP toxins is an important part for promoting safety supervision system of sea food in China. In this paper, effects of post-column derivatization condition on reaction fluorescent intensity and mobile phase condition on separation of gonyautoxins (GTXs) base on HPLC were discussed. Analysis of post-column derivatization showed the effects of pH of reaction solution, reaction temperature, formamide concentration and ammonium formate concentration on fluorescent sensitivity of individual GTX compounds were remarkable different. Considering the relationship between reaction fluorescent intensity and reaction conditions, the optimum post-column derivatization conditions were defined as: reaction solution mixtured by 70 mmol/L periodic acid and 0.12 mol/L potassium hydrate, 0.75 mol/L formamide of 20% ammonium formate solution, with pH of 7.3～7.5, flow rate of 0.8 ml/min, and reaction temperature of 50℃. Analysis of mobile phase showed the effects of methanol concentration, sodium heptanesulfonate concentration, pH, and phosphate concentration on separation of GTXs were all obvious, and the optimum mobile phase conditions were determined as: the concentrations of methanol, sodium heptanesulfonate, phosphate were 1%, 1.5 mmol/L, 10 mmol/L respectively, with pH of 7.3. Comparison to HPLC of oshima method, the response rate of GTX1, GTX3 and GTX4 increased to 4.7, 3.4 and 3.8 times, respectively, beside that of GTX2.

中图分类号:

Q6573

姜沛宏章超桦秦小明卢虹玉林华娟梁颖瑜. 柱后衍生高效液相色谱法检测膝沟藻毒素[J]. 食品科学, 2013, 34(2): 170-174.

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