壳聚糖纳米粒修饰的开管毛细管电色谱柱对生物胺的分析

Analysis of Amines by Open Tubular Capillary Electrochromatography Column Modified with Nano-Chitosan Particles

目的建立应用壳聚糖纳米粒修饰的开管毛细管电色谱柱对生物胺的分析检测方法。方法通过静电作用将壳聚糖纳米粒修饰于毛细管内壁;优化应用该柱对8种生物胺包括组胺、色胺、酪胺、5-羟色胺、多巴胺、苯乙胺、腐胺和尸胺的分离条件,通过测定日内、日间和柱间的相对标准偏差（RSD）考察该方法的重现性;通过加标-计回收率的方法检测鱼露中生物胺的含量。结果在pH 6.7的50mmol/L的磷酸盐缓冲液、10kV电压的条件下,8种生物胺在15min内得到完全分离,柱效范围为52 397~95 302plates/m,并且日内、日间和柱间的迁移时间的RSD分别小于1.8%,3.6%和8.0%,峰面积RSD分别小于4.0%,6.7%和13.0%;检测定量限为1.4~5.0mg/L;回收率为82%~112%。结论壳聚糖纳米粒子修饰的开管毛细管电色谱柱的应用提高了分析生物胺的效果,并可用于实际样品中生物胺的检测。

Objective To develop a capillary electrochromatography method with open tubular capillary column modified by nano-chitosan particles for determination of biogenic amines,including histamine,tryptamine,tyramine,5-hydroxytryptamine,dopamine,phenethyl-amine,putrescine and cadaverine. Method The chitosan nano-particle were coated onto the inner wall of fused silica capillary via electrostatic adsorption;optimal separation conditions of 8amines were investigated,and the reproducibility of the method was also assessed via RSD of intra-day,inter-day and column to column;recoveries of 8amines were determined after standard substance addition. Result Under the condition of 50mmol/L phosphate buffer solution at pH 6.7with 10 kV voltage,8biogenic amines were separated completely within 15 min,and the column efficiency range was 52 397~95 302plates/m. The RSD of retention times were lower than 1.8%,3.6%and 8.0%for intra-day,inter-day and column to column,respectively,and correspondingly the RSD of peak areas were lower than 4.0%,6.7% and 13.0%,respectively. This method was applied in biogenic amine analysis in fish sauce,and the average recoveries were in the range of82%~112%. The proposed method allowed amines to be determined with limit of detection of1.4~5.0mg/L. Conclusion Efficient separation of amines was performed by CEC using nano-CS coating column,which can be used in the analysis of biogenic amines in fish sauce.