超高效液相色谱紫外-柱前衍生法测定肉制品中的9种生物胺

Determination of 9 kinds of biogenic amines in meat products by ultra performance liquid chromatography with ultraviolet-pre-column derivatization

中文关键词:  超高效液相色谱法  生物胺  样品提取  衍生条件  火腿肠  肉制品

英文关键词:ultra performance liquid chromatography  biogenic amine  sample extraction  derivatization conditions  ham sausage  meat products

中文摘要:

      目的 建立超高效液相色谱紫外-柱前衍生法测定肉制品中的9种生物胺的分析方法。方法 实验选取西式火腿肠为样品, 以三氯乙酸/高氯酸混合溶液对样品中生物胺进行提取, 调节提取液于pH 10的碱性环境下, 使用1 mL的10 mg/mL丹磺酰氯溶液进行衍生化处理, 60 ℃水浴衍生15 min, 100 μL氢氧化钠中止衍生, 乙酸乙酯进行最终萃取。使用超高效液相色谱仪紫外配二极管阵列检测器进行测定, 流动相为1%乙酸乙腈-10 mmol/L乙酸铵溶液; ACQUITY BEH C18 (100 mm×2.1 mm, 1.7 μm)色谱柱进行分析。结果 在线性范围5~250 mg/kg内9种生物胺的相关系数均大于0.9990, 检出限范围为0.2~0.5 mg/kg, 定量限范围为0.5~ 1.5 mg/kg, 9种生物胺的日内精密度相对标准偏差(relative standard deviations, RSDs)范围为1.58%~5.21%, 回收率均大于等于85%。结论 本研究所建方法具有良好的准确性、精密度、灵敏度, 可为我国肉制品中生物胺的检测提供参考依据。

英文摘要:

      Objective To establish an analytical method for the determination of 9 kinds of biogenic amines in meat products by ultra performance liquid chromatography with ultraviolet-pre-column derivatization. Methods In the experiment, western-style ham sausage was taken as the sample, and the biogenic amines in the sample were extracted with the mixed solution of trichloroacetic acid and perchloric acid, the extracting solution was adjusted to the alkaline environment at pH 10 for derivatization using 1 mL of 10 mg/mL dansyl chloride solution, the derivatization was carried out in a water bath at 60 ℃ for 15 min, and the derivatization was stopped with 100 μL sodium hydroxide, and the final extraction was conducted with ethyl acetate. The measurement was performed using an ultra performance liquid chromatographic instrument with an ultraviolet diode array detector, and the mobile phase was 1% acetonitrile-10 mmol/L ammonium acetate solution, ACQUITY BEH C18 (100 mm×2.1 mm, 1.7 μm) column was used for analysis. Results The correlation coefficients were greater than 0.9990 in the linear range of 5?250 mg/kg, the detection limits range of 0.2?0.5 mg/kg, the limits of quantitation range of 0.5?1.5 mg/kg, and the relative standard deviations (RSDs) of intra-day precision of the 9 kinds of biogenic amines were 1.58%?5.21%, and the recoveries were all greater than or equal to 85%. Conclusion The established method has good accuracy, precision and sensitivity, and can provide reference for the determination of biogenic amines in meat products in China.