固相萃取-高效液相色谱法同时测定羊肉制品中9种杂环胺

建立了同时测定羊肉制品中9种杂环胺的固相萃取-高效液相色谱(SPE-HPLC)分析方法。样品经2 mol/L NaOH超声提取、80 mL二氯甲烷液-液萃取,利用阳离子交换柱(MCX柱)净化和富集后进行HPLC分析。采用反相C18色谱柱,以乙腈和0.01 mol/L磷酸溶液(用三乙胺调整pH 3.6)为流动相对杂环胺进行梯度洗脱,使用二极管阵列检测器(DAD)分别在228 nm(2-氨基-9H-吡啶并［2,3-b］吲哚(AaC)、2-氨基-3-甲基-9H-吡啶并［2,3-b］吲哚(MeAaC))、253 nm(2-氨基-3-甲基咪唑并［4,5-f］喹啉(IQ)、1-甲基-9H-吡啶并［3,4-b］吲哚(Harman)、2-9H-吡啶并［3,4-b］吲哚(Norharman))、263 nm(2-氨基-3,8-二甲基咪唑并［4,5-f］喹喔啉(MeIQx)、2-氨基-3,4,8-三甲基咪唑并［4,5-f］喹喔啉(4,8-DiMeIQx)、3-氨基-1-甲基-5H-吡啶并［4,3-b］吲哚(Trp-p-2))、321 nm(2-氨基-1-甲基-6-苯基咪唑并［4,5-b］吡啶(PhIP))处进行检测。实验结果表明,9种杂环胺分离效果良好,回收率为50.27%～94.77%(n=6),相对标准偏差为0.08%～4.42%;通过全波长扫描,确定检出限(以信噪比(S/N)=3计)为1.6～41.0 μg/L。该方法操作简单,结果准确,重现性好,可用于同时检测羊肉制品中9种杂环胺的含量。

Simultaneous determination of nine heterocyclic aromatic amines in mutton products by solid phase extraction-high performance liquid chromatography

method was established for the simultaneous determination of nine heterocyclic aromatic amines (HAAs) in mutton products by solid phase extraction-high performance liquid chromatography (SPE-HPLC). As a result, the sample was prepared by ultrasound in 2 mol/L NaOH, and dichloromethane was selected as the extraction solvent. The extract was purified and concentrated with an MCX SPE column. The chromatographic separation was achieved on a reverse-phase C18 column by gradient elution using 0.01 mol/L phosphoric acid (adjusted to pH 3.6 by triethylamine) and acetonitrile, detected with a diode array detector (DAD) at 228 nm for 2-amino-9H-pyrido［2,3-b］indole (AaC) and 2-amino-3-methyl-9H-pyrido［2,3-b］indole (MeAaC); 253 nm for 2-amino-3-methylimidazo［4,5-f］quinoline (IQ), 1-methyl-9H-pyrido［3,4-b］indole (Harman), 9H-pyrido［3,4-b］indole (Norharman); 263 nm for 2-amino-3,8-dimethylimidazo［4,5-f］quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo［4,5-f］quinoxaline (4,8-DiMeIQx), 3-amino-1-methyl-5H-pyrido［4,3-b］indole (Trp-p-2); 321 nm for 2-amino-1-methyl-6-phenylimidazo［4,5-b］pyridine (PhIP). The results showed that the nine HAAs can be separated efficiently. The recoveries (n=6) of the nine HAAs spiked in meat were between 50.27% and 94.77% with the relative standard deviations of 0.08%~4.42%. The limits of detection of this method were 1.6~41.0 μg/L for the nine HAAs. The method is simple, accurate, rapid and repeatable. It can be used for the simultaneous determination of the nine HAAs in mutton products.