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