6种农业特种养殖动物肉中18种氨基酸含量分析

Content s of 18 Amino Acids in Meat from 6 Species of Special Farm Animals

针对目前国内新兴的6种农业特种养殖动物,即蓝孔雀、眼镜蛇、滑鼠蛇、海蛇、海龙和海星,探讨6种动物肉中18种氨基酸含量的检测方法。运用全自动氨基酸分析仪法测定上述6种动物的18种氨基酸含量,样品经90%乙醇沉淀蛋白质后用6 mol/L盐酸溶液水解22 h后离心,上清液氮吹干,用样品稀释液复溶,过滤膜上机检测。以柠檬酸钠和氢氧化钠溶液为流动相进行梯度洗脱,荧光检测器测定,除脯氨酸(Pro)检测波长为440 nm外,其余17种氨基酸检测波长均为570 nm。结果表明:除胱氨酸质量浓度范围为1.6～8.0 mg/L、牛磺酸为2.0～10.0 mg/L外,其余16种氨基酸质量浓度范围为3.2～16.0 mg/L时线性良好(r=0.998 5～0.999 9);方法检出限为0.04～0.42 mg/L;低质量浓度(16种氨基酸质量浓度为6.4 mg/L,胱氨酸为3.2 mg/L,牛磺酸为4.0 mg/L)条件下的加标回收率为96.11%～109.76%,相对标准偏差(relative standard deviation,RSD)为0.14%～1.62%,高质量浓度(16种氨基酸质量浓度为12.8 mg/L,胱氨酸为6.4 mg/L,牛磺酸为8.0 mg/L)条件下的加标回收率为96.12%～104.13%,RSD为0.11%～1.88%;精密度RSD为0.08%～0.94%,稳定性RSD为0.16%～0.97%,重复性RSD为0.09%～0.91%。表明该方法快速易行、准确可靠,可用于蓝孔雀、眼镜蛇、滑鼠蛇、海蛇、海龙和海星的检验方法优化。

In this paper, the contents of 18 amino acids in meat from 6 newly developed special farm animals, namely blue peacock, cobra, mouse snake, sea snake, sea dragon and sea star, were determined with an automatic amino acid analyzer.After protein precipitation with 90% ethanol, the samples were hydrolyzed with 6 mol/L hydrochloric acid solution for 22 h and then centrifuged. The supernatant was blown to dryness with nitrogen and the residue was re-dissolved and ?ltrated before being injected and analyzed. A mixture of sodium citrate and sodium hydroxide solutions was used as the mobile phase for gradient elution and ?uorescence detection was performed at 440 nm for Pro and at 570 nm for 17 other amino acids. The results showed that the linear ranges were 1.6–8.0 mg/L for cystine, 2.0–10.0 mg/L for taurine, and 3.2–16.0 mg/L for other amino acids(r = 0.998 5–0.999 9). The detection limits of the method were 0.04–0.42 mg/L. Recoveries from samples spiked at 3.2 mg/L for cystine, 4.0 mg/L for taurine, and 6.4 mg/L for other amino acids were in the range between 96.11% and109.76% with relative standard deviations(RSDs) between 0.14% and 1.62%. Recoveries from samples spiked at 6.4 mg/L for cystine, 8.0 mg/L for taurine, and 12.8 mg/L for other amino acids were in the range between 96.12% and 104.13% with RSDs were 0.11%-1.88%. The precision, stability and repeatability RSDs were 0.08%–0.94%, 0.16%–0.97% and 0.09%–0.91%,respectively. This method was rapid, accurate and reliable, and could be used to determine amino acids in blue peacock, cobra,mouse snake, sea snake, sea dragon and sea star.