两种柱前衍生RP-HPLC法测定酱腌菜中生物胺的比较

Comparison on Two Pre-column Derivatization Methods for RP-HPLC Determination of Biogenic Amines in Pickled Vegetable

本研究分别采用4-氟-3-硝基三氟甲苯(FNBT)柱前衍生RP-HPLC紫外检测分析方法(简称FNBT衍生法)和丹磺酰氯柱前衍生RP-HPLC紫外检测分析方法(简称DNS-Cl衍生法)测定酱腌菜中生物胺含量,并比较这两种方法对酱腌菜中8种生物胺(组胺、色胺、β-苯乙胺、腐胺、酪胺、尸胺、亚精胺及精胺)检测的差异。结果表明,在浓度范围1~100mg/L内,两种方法对8种生物胺的检测呈现良好的线性相关性,相关系数R^2均大于0.999,FNBT衍生法的检出限(0.016~0.071mg/L)优于DNS-Cl衍生法(0.066~0.191 mg/L)。方法的精确度试验表明:FNBT衍生法的加标平均回收率(92.87%~100.18%)略优于DNS-Cl衍生法的加标平均回收率(83.52%~102.33%),两种方法的测量精密度RSD%均小于5%。DNS-Cl衍生法衍生试剂消耗大、操作繁琐,对生物胺检测的灵敏度低,而FNBT衍生法衍生试剂消耗少、操作简便、快速、灵敏度及准确性更高,适用于大批量食品中微量生物胺的快速、准确分析。用两种方法检测酱腌菜中8种生物胺的含量存在显著差异(p<0.05)。

This study used the 4-fluoro-3-nitrobenzotrifluoride（FNBT） pre-column derivatization RP-HPLC analysis method with UV detector（abbreviated as FNBT method） and the dansyl chloride pre-column RP-HPLC analysis method with UV detector（abbreviated as DNS-Cl method） to determine biogenic amines in pickled vegetable, and compared the differences between the two methods in 8 types of biogenic amines in pickled vegetable（histamine, tryptamine, guanidine-phenylethylamine, putrescine, tyramine, cadaverine, spermidine and spermine）. Results showed that in the concentration range of 1-100 mg/L, the two methods showed a good linear with the detection of the eight biogenic amines with the correlation coefficient R2 greater than 0.999 and detection limit of the FNBT derivatization method（0.016-0.071 mg/L） lower than that of DNS-Cl derivatization method（0.066-0.191 mg/L）. The accuracy test of the methods showed that the average recovery of the FNBT derivatization method（92.87%-100.18%） were slightly better than that of DNS-Cl derivatization method（83.52%-102.33%）, with the precision（RSD%） of the two different methods less than 5%. DNS-Cl derivatization method requires a large amount of expensive reagents and cumbersome operation with low sensitivity to biogenic amines analysed, whilst the FNBT derivatization method allows simple, rapid and accurate analyses of eight biogenic amines with low consumption of derivatization reagents and higher sensitivity for biogenic amines. FNBT derivatization method is more suitable for rapid and accurate analysis of trace biogenic amines in large quantities of food. There were significant differences in the levels of eight biogenic amines in pickled vegetables analyzed by two methods（P〈0.05）.