Effect of Refrigeration Time on Changes in Biogenic Amine Contents in Large Yellow Croaker and Pomfret

To establish a high-performance liquid chromatography (HPLC) method for the determination of nine biogenic amines and to study the changes of biogenic amine contents in large yellow croaker and pomfret during cold storage.

The samples were extracted with perchloric acid solution and derivatized with dansylchloride, followed by addition of ammonia water to terminate the derivatization reaction, standing in the dark and making up the volume with acetonitrile. Chromatographic separation was performed on an Agilent Eclipse XDB-C₁₈ column (4.6 mm ×250 mm, 5 μm) with gradient elution using a mobile phase consisting of 5 mmol/L ammonium acetate in water and acetonitrile at a flow rate of 0.8 mL/min. Detection was carried out using a diode array detector at a wavelength of 254 nm.

The proposed method effectively separated the nine biogenic amines within 35 min. The calibration curve for the mixed standard of biogenic amines exhibited good linearity in the concentration range of 0.1–80.0 mg/L with correlation coefficients greater than 0.995. The limit of detection (LOD) was 0.5 mg/kg, and the limit of quantitation (LOQ) was 1.0 mg/kg. The recoveries of spiked samples ranged from 75.70% to 105.40% with relative standard deviation (RSD) ≤ 5% (n = 6).

This method is simple, accurate and reliable. Furthermore, it was used to determine the changes in biogenic amine contents of large yellow croaker and pomfret during refrigerated storage. The results showed that the contents of putrescine, cadaverine and tyramine changed significantly, while the changes in spermidine and spermine contents were gentle. After 30 days of refrigeration, the contents of putrescine, cadaverine, and tyramine increased by 161, 143, and 49.2 mg/kg in large yellow croaker, respectively. The contents of putrescine, cadaverine, and tyramine increased by 168, 192, and 221 mg/kg in pomfret, respectively. The above findings indicate that under the same storage conditions, pomfret was more likely to produce tyramine than large yellow croaker, and tyramine content changed more significantly than did putrescine and cadaverine contents.