柱前衍生反相高效液相色谱法测定狭叶红景天中氨基酸含量
目的 建立柱前衍生反相高效液相色谱法(reversed-phase high-performance liquid chromatography, RP-HPLC)测定狭叶红景天中游离及水解氨基酸种类及含量。方法 以异硫氰酸苯酯为柱前衍生化试剂; 色谱柱采用Ultimate Amino Acid氨基酸柱 (4.6 mm×250 mm, 5 μm); 流动相A为80%(V:V)的乙腈-水溶液, 流动相B为7%(V:V)的乙腈-CH3COONa溶液(pH=6.5); 检测波长为254 nm。结果 21种氨基酸在0.0019~2.700 mmol/L内呈良好的线性关系(r2>0.9990)。游离氨基酸平均回收率在97.80%~116.82%之间, RSD在1.93%~2.47%之间(n=6); 水解氨基酸平均回收率在99.26%~111.23%之间, RSD在1.70%~2.23%之间(n=6)。结论 柱前衍生RP-HPLC法灵敏、准确, 具有良好的重复性和稳定性, 适合对狭叶红景天中氨基酸含量的测定。
Objective To determine the amino acid content in Rhodiola kirilowii (Regel) Maxim by pre-column derivatization reversed-phase high performance liquid chromatography (RP-HPLC). Methods The samples were derived with phenyl isothiocyanate (PITC). The column was of specific column from Ultimate Amino Acid (4.6 mm×250 mm, 5 μm). Mobile phase A was a mixture of acetonitrile and water (80%, V:V) and mobile phase B was acetonitrile (7%, V:V)-CH3COONa(pH=6.5). Wavelength of ultraviolet detector was set at 254 nm. Results The linear range of 21 kinds of amino acids was 0.0019~2.7000 mmol/L (r2>0.9990). The average recovery rates of 21 kinds of free amino acids ranged from 97.80% to 116.82% (RSD=1.93%~2.47%, n=6) and the average recovery of rates of 21 hydrolysis amino acids ranged from 99.26% to 111.23% (RSD=1.70%~2.23%, n=6). Conclusion This method is sensitive, accurate, repeatable and stable for the determination of amino acids in Rhodiola kirilowii (Regel) Maxim.
标题：柱前衍生反相高效液相色谱法测定狭叶红景天中氨基酸含量
英文标题：Determination of amino acid content in Rhodiola kirilowii (Regel) Maxim by pre-column derivatization reversed-phase high performance
作者：
赵岩 吉林农业大学中药材学院
赵天琦 吉林农业大学中药材学院
陆珞 吉林省食品药品审评中心
阎墨 吉林省食品药品审评中心
郜玉钢 吉林农业大学中药材学院
何忠梅 吉林农业大学中药材学院
张连学 吉林农业大学中药材学院
中文关键词:狭叶红景天,柱前衍生,氨基酸,高效液相色谱法,
英文关键词:Rhodiola kirilowii (Regel) Maxim,pre-column derivatization,amino acid,high performance liquid chromatography,
发表日期：2015-03-30
- 文件大小:
- 716.45 KB
- 下载次数:
- 60
-
高速下载
|
|
