水产品中腐胺和亚精胺的快速分离与检测

Rapid Separation and Determination of Putrescine and Spermidine in Aquatic Products

基于腐胺、亚精胺能够与Ru（bpy）3^2＋发生电化学反应,显著增强Ru（bpy）3^2＋的发光强度,通过毛细管电泳-电化学发光联用,对水产品中的腐胺和亚精胺进行快速分离检测。经试验得到最优条件：检测电位1.15 V;Ru（bpy）3^2＋浓度5 mmol/L（溶于pH 7.50磷酸盐缓冲液）;运行高压16 kV;进样电压10 kV,进样时间10 s;运行缓冲液使用pH 7.00,50 mmol/L磷酸盐缓冲液。在此条件下,5×10^-5-1×10^-2mg/L的腐胺标准品浓度与发光强度呈良好的线性关系,线性方程为I=57.524c＋292.88,相关系数r=0.9888（n=5）,最低检出限（S/N=3）为1×10^-7mg/L。1×10^-5-1×10^-3mg/L的亚精胺标准品浓度与发光强度呈良好的线性关系,线性方程I=7 581.2c＋1 155.6,相关系数r=0.9967（n=5）,最低检出限（S/N=3）为1×10^-8 mg/L。在最优条件下对样品中腐胺、亚精胺进行分离、检测,4种样品中均未发现亚精胺,沙丁鱼、巴浪鱼中未发现腐胺。带鱼中腐胺的峰高、迁移时间的相对标准偏差分别为2.35%,0.71%,加标回收率,102.76%-103.2%;秋刀鱼中腐胺的峰高、迁移时间的相对标准偏差分别为3.41%,0.52%,加标回收率96.1%-117.76%。

A sensitive, simple and rapid CE-ECL method is established for the determination of putrescine and spermine in aquatic product. This method is based on the electrochemical reaction of putrescine, spermidine and Ru（bpy）3^2 ＋occurred on the platinum electrode. Experimental conditions for the biogenic amines＇ separation and detections were optimized in details. The optimal conditions were：a detection potential of 1.15 V; 50 mmol, pH 7.00 phosphate buffer in the detection cell containing 5 mmol Ru（bpy）3^2＋（pH=7.00）; a separation voltage of 15 kV; a sample injection at 10 kV ×10 s; Under these conditions, the ECL intensity was linear with the concentrations of putrescine over the ranges from 5×10^-5-1×10^-2mg/L. The linear equation for I = 57.524 c ＋ 292.88, correlation coefficient r = 0.9888（n = 5） with detection limit（S/N = 3） of 1×10^-7mg/L. For spermidine, the ECL intensity was linear with the concentrations of sper mine over the ranges from 1×10^-5-1×10^-3mg/L. The linear equation for I = 7 581.2c ＋ 1 155.6, correlation coefficient r =0.9967（n = 5） with detection limit（S/N = 3） of 1×10^-8mg/L. Under optimal conditions, no spermidine is detected in all four samples and there is none putrescine detected in the sardine and decapterus maruadsi samples. For the determination of putrescine, in the hairtail samples, the relative standard deviation（RSD） of peak height and migration time are2.35% and 0.71%, recovery is 102.76%-103.2%. In the saury samples, the relative standard deviation（RSD） of peak height and migration time are 3.41% and 0.52%, recovery is 96.1%-117.76%.