Improvement of National Standard Method for Determination of 8 Biogenic Amines in Livestock and Poultry Meat by High Performance Liquid Chromatography

In view of the cumbersome operation and the long detection time (about 6 h) of the national standard method of GB 5009.208-2016, extraction and derivatization methods were improved, and the proposed method was applied to the determination of 8 biogenic amines in livestock and poultry meat, including tryptamine,cadaverine, β-phenethylamine, tyramine, spermidine, putrescine, histamine and spermine. The minced sample (10 g)JP was taken and mixed with 20 mL of 50 g·L^-1 trichloroacetic acid solution, 10 mL of n-hexane and 1.0 mL of 1 000 mg·L^-1 1,7-diaminoheptane internal standard solution by vortex for 15 min. After ultrasonic extraction for 15 min, and centrifugation for 5 min, 0.5 mL of the supernatant was taken and placed into 10 mL-colorimetric tube. 100 μL of 2 mol·L^-1 sodium hydroxide solution, 300 μL of saturated sodium bicarbonate solution, and 2.0 mL of 10 g·L^-1 dansyl chloride derivative solution were added subsequently. The mixed solution was mixed by shaking, reacted at 60℃ for 15 min, and 100 μL of ammonia water was added for reaction at 60℃ for 15 min to stop the derivatization reaction. The solution obtained was diluted to 5 mL with acetonitrile, and passed through a 0.22 μm filter membrane. The filtrate was introduced into high performance liquid chromatograph, separated on Phenomenex Kinetex column with methanol-water system at different volume ratios for gradient elution, and analyzed with diode array detector. It was shown that the sample could be analyzed within 1.5 h, and the mass concentrations of the 8 biogenic amines were all linearly related with ratios of the peak areas between the target and the internal standard within 5.00-250 mg·L^-1, with detection limits (3S/N) of 4-21 μg·kg^-1. Tests for spiked recovery and intra-day and inter-day precision were made on the actual samples, giving recoveries in the range of 80.4%-95.6%, and RSDs of the determined values were 1.9%-5.1% (for intra-day precision test, n=6) and 2.3%-6.2% (for inter-day precision test, n=5), respectively. The proposed method was used for the analysis of 20 batches of livestock and poultry meat. Except for histamine, the other 7 biogenic amines were detected to varying degrees.