矛头蝮蛇蛇毒中游离氨基酸测定与分析
Determination and analysis of free amino acids in snake venom
薛雁 1李秀琳 1李秀娜 1丁忠福 1崔亮亮1
作者信息
- 1. 辽宁远大诺康生物制药有限公司,辽宁 沈阳 110171
- 折叠
摘要
目的 建立一种同时测定蛇毒中18种游离氨基酸含量的方法.方法 采用膜过滤法制备蛇毒游离氨基酸样品溶液.以异硫氰酸苯酯(PITC)柱前衍生化后进行HPLC分析,采用Ultimate LP-C18色谱柱,流动相A 0.05 mol/L乙酸钠,流动相B为甲醇:乙腈:水=40:40:20,梯度洗脱,流速1.0 mL/min,柱温35℃,检测波长254 nm.结果 18种氨基酸线性相关系数均大于0.99,加样回收率在74.59%~110.62%.蛇毒中含有17种氨基酸,氨基酸的总含量为0.2%.结论 该方法精密度高、重复性好、结果准确可靠,能用于蛇毒中及其相关产品中氨基酸含量测定.
Abstract
Objective To establish a method for simulataneous determination of 18 free amino acids in snake venom. Methods Preparation of free amino acid samples by membrane. HPLC analysis was performed after derivatization by using phenyl isothiocyanate (PITC) as a derivative reagent, samples were analyzed on Ultimate LP-C18 column with gradient elution column of 0.05 mol/L sodium acetate buffer and methanol-acetonitrile- water (40:40:20), and current speed was1.0 mL/min, and the column temperature was set at 35℃, and detection wavelength was 254nm. Results The 18 kinds of amino acids showed a good linearity with the correlation coefficients ≥0.99. The recovery rate was 74.59%~110.62%. Snake venom contained 17 kinds of amino acids, the total content of amino acids was 0.2%. Conclusion The method was accurate, reproducible and reliable, and can be used for the determination of amino acids in snake venom and related products.
关键词
蛇毒/氨基酸/异硫氰酸苯酯/HPLCKey words
snake venom/amino acids/phenyl isothiocyanate/HPLC引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据