固相萃取-高效液相色谱法同时测定传统禽肉制品中的9种杂环胺类化合物

建立了固相萃取-高效液相色谱同时测定传统禽肉制品中9种杂环胺类化合物(HAAs)(包括2-氨基-3-甲基咪唑并［4,5-f］喹啉、2-氨基-3,4-二甲基咪唑并［4,5-f］喹啉、2-氨基-3,8-二甲基咪唑并［4,5-f］喹喔啉、2-氨基-3,4,8-三甲基咪唑并［4,5-f］喹喔啉、2-氨基-1-甲基-6-苯基-咪唑并［4,5-b］吡啶、3-氨基-1-甲基-5H-吡啶并［4,3-b］吲哚、3-氨基-1,4-二甲基-5H-吡啶并［4,3-b］吲哚、9H-吡啶并［4,3-b］吲哚、1-甲基-9H-吡啶并［4,3-b］吲哚)含量的分析方法。经过条件优化,肉样选用乙酸乙酯进行提取,提取液经丙基磺酸(PRS)和C18固相萃取小柱净化,采用TSK-gel ODS-80TM色谱柱,以乙腈和0.05 mol/L醋酸-醋酸铵缓冲液(pH 3.4)为流动相进行梯度洗脱分离,紫外-荧光检测器串联方式对目标化合物进行检测。通过波长扫描,确定紫外检测波长为263 nm,荧光激发波长/发射波长随时间切换程序为: 0～21 min, 300 nm/440 nm; 21～23.8 min, 315 nm/410 nm; 23.8～35 min, 265 nm/410 nm。在上述条件下,9种HAAs在35 min内实现基线分离。3个加标水平的平均回收率为60.47%～90.55%(n=6),相对标准偏差(RSD)为0.49%～9.74%(n=6),检出限(以信噪比为3计)为0.1～3.6 μg/kg。该方法简便快速、结果准确、灵敏度高,可作为测定传统禽肉制品中多种杂环胺类化合物的有效方法。

Simultaneous determination of 9 heterocyclic aromatic amine in poultry products by solid-phase extraction-high performance liquid chromatography

A method was developed for the simultaneous determination of 9 heterocyclic aromatic amines (HAAs) including 2-amino-3-methylimidazo[4,5-f] quinoline (IQ), 2-amino-3,4-dimethylimidazo[4, 5-f] quinoline (MeIQ), 2-amino-3, 8-dimethylimidazo [4, 5-f] quinoxaline (MeIQx), 2-amino-3, 4, 8-trimethyl-imidazo [4,5-f quinoxaline (4,8-DiMeIQx), 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP), 3-amino-1 -methyl-5H-pyrido [4, 3-b] indole (Trp-p-2), 3-amino-1, 4-dimethyl-5H-pyrido [4,3-b] indole (Trp-p-1), 9H-pyrido [3,4-b] indole (Norharman), 1-methyl-9H-pyrido 3, 4-b] indole (Harman)) in poultry products using solid-phase extraction-high performance liquid chromatography (SPE-HPLC). The performance of 3 different organic extraction solutions, 2 solid phase extraction (SPE) procedures, three different reversed-phase columns and five different mobile phases were tested for optimizing separation conditions of the 9 HAAs from poultry products. In the end, ethyl acetate was selected as the extraction solution, and the extract was purified with propylsulfonic acid silica (PRS) and C18 SPE columns. The analysis was performed on a TSK-gel ODS-80TM column using a gradient elution with the mobile phases of 0.05 mol/L aqueous acetic acid-ammonium acetate buffer (pH 3.4) and acetonitrile. The results showed that the average recoveries (n = 6) of the 9 HAAs spiked in meat samples at 3 levels ranged from 60.47% to 90.55% with the relative standard deviations (RSDs) between 0.49% and 9.74%, and the limits of detection (LODs, S/N = 3) were in the range of 0.1 - 3.6 microg/kg. The method is simple, rapid, accurate and sensitive enough for the analysis of HAAs in poultry products.