冷藏时间对大黄鱼、鲳鱼中生物胺含量变化的影响

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

目的:建立一种以高效液相色谱法测定9种生物胺含量的方法,研究冷藏时间对大黄鱼、鲳鱼生物胺含量的影响。方法:大黄鱼、鲳鱼样品经高氯酸溶液溶解提取,丹磺酰氯衍生剂衍生,氨水终止衍生反应,避光静置,乙腈定容。利用Agilent Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm)分离,以5 mmol/L乙酸铵和乙腈为流动相进行梯度洗脱,流速为0.8 mL/min,二极管阵列检测器在波长254 nm下检测。结果:此方法可将9种生物胺在35 min内有效分离;在0.1~80.0 mg/L范围内,9种生物胺混合标准溶液质量浓度与峰面积呈良好的线性关系,相关系数均大于0.995;方法检出限为0.5 mg/kg,定量限为1.0 mg/kg;加标回收率介于75.70%~105.40%之间,相对标准偏差≤5%(n=6)。结论:所建立的方法能够对大黄鱼、鲳鱼中9种生物胺含量进行简便、准确、可靠的分析;利用该方法研究大黄鱼和鲳鱼中生物胺含量随冷藏时间的变化规律,结果表明,腐胺、尸胺和酪胺含量变化较明显,亚精胺和精胺含量变化平缓,冷藏30 d的大黄鱼,腐胺、尸胺和酪胺含量分别增加161、143、49.2 mg/kg,冷藏30 d的鲳鱼,腐胺、尸胺和酪胺含量分别增加168、192、221 mg/kg,同一贮藏条件下,鲳鱼比大黄鱼更易产生酪胺,含量变化最明显。

Objective: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.Methods: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,5μm)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.Results: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).Conclusion: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.