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

Determination of amino acid content in Rhodiola kirilowii (Regel) Maxim by pre-column derivatization reversed-phase high performance

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

中文摘要:

      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。

英文摘要:

      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.