PICO·TAG法测定王锦蛇中不同部位游离氨基酸含量

PICO·TAG Determination of Amino Acid Content of Different Parts in Elaphe Carinat

关键词:  王锦蛇  氨基酸  高效液相色谱法  PITC柱前衍生

Key words:  Elaphe carinat  amino acids  HPLC  pre-column derivated by PITC

摘要:

目的 建立PITC(异硫氰酸苯酯)柱前衍生化反相高效液相色谱法测定王锦蛇(菜花蛇)中蛇肉、蛇心、蛇肺、蛇肠中的17种游离氨基酸含量，进一步开发和利用王锦蛇的食用与药用价值。方法 Waters PICO·TAG^TM游离氨基酸分析柱(3.9 mm×150 mm，5 μm)，以醋酸钠缓冲液为流动相A，乙腈-水(60∶40)为流动相B梯度洗脱，254 nm紫外条件下检测分析。结果 20 min内17种氨基酸均得到较好的分离；17种氨基酸在各自的浓度范围内线性关系良好，r在0.997 6～0.999 9之间，平均加样回收率为98.3%～107.5%。结论 该方法操作简便，准确、可靠，重复性好，适用于王锦蛇中蛇肉、蛇心、蛇肺、蛇肠的氨基酸含量测定，为王锦蛇资源的进一步开发利用和保护提供了实验依据。

Abstract:

OBJECTIVE To establish a reversed-phase high-performance liquid chromatographic method derivated by PITC (phenylisothiocyanate) pre-column for the determination of 17 amino acids content of fresh, heart, lung and intestines in Elaphe carinat (Cauliflower), and to develop the edible and medical value of Elaphe carinat further. METHODS Using Waters PICO·TAG^TM column(3.9 mm×150 mm, 5 μm), gradient elution with sodium acetate buffer as mobile phase A, acetonitrile-water (60∶40) as mobile phase B, detection wavelength of 254 nm. RESULTS All of 17 amino acids were well separated within 20 min; 17 amino acids had a good linear relationship in their respective concentration ranges (0.997 6<r<0.999 9). The average recoveries were in 98.3%-107.5%. CONCLUTION This method is simple, accurate and reliable with a good reproducibility and can be used to determine amino acids content of snake fresh, heart, lung and intestines in Elaphe carinat, which provides experimental evidence of the further development, usage and protection of the Elaphe carinat resource.