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目的采用柱前衍生反相高效液相色谱法对草本水杨梅叶中氨基酸进行测定。方法样品以异硫氰酸苯酯(PITC)为衍生化试剂,与草本水杨梅叶中氨基酸柱前衍生,采用Ultimate Amino Acid氨基酸分析专用柱(4.6 mm×250 mm,5μm);流动相:A为乙腈-水(80:20,V:V),B为乙腈-三水合乙酸钠缓冲溶液(7:93,V:V,p H 6.5),梯度洗脱;柱温35℃,流速1.0 m L/min,检测波长为254 nm。结果 20种氨基酸在0.039~1.250 mmol/L内呈良好的线性关系(r2>0.9996)。20种游离氨基酸平均回收率在96.51%~108.06%之间,RSD在0.36%~2.08%之间(n=6);水解氨基酸平均回收率在95.29%~102.13%之间,RSD在0.85%~2.45%之间(n=6)。结论该方法可用于草本水杨梅中氨基酸的检测,其灵敏度高且准确,具有良好的重复性及稳定性。
OBJECTIVE To determine the amino acids in leaves of Myrica rubra by precolumn derivatization reversed-phase high performance liquid chromatography. Methods PITC was used as a derivatization reagent to derivatize the amino acids in Herba pakaemirensis leaves. Ultimate Amino Acid column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of A Was acetonitrile-water (80:20, V: V) and B was acetonitrile-sodium acetate trihydrate buffer solution (7:93, V: V, p H 6.5) L / min, detection wavelength of 254 nm. Results The 20 amino acids showed good linearity (r2> 0.9996) in the range of 0.039-1.250 mmol / L. The average recoveries of 20 kinds of free amino acids ranged from 96.51% to 108.06%, RSD ranged from 0.36% to 2.08% (n = 6). The average recoveries of hydrolyzed amino acids ranged from 95.29% to 102.13% with RSD from 0.85% 2.45% (n = 6). Conclusion The method can be used for the determination of amino acids in Herba Maywendra. Its sensitivity is high and accurate, with good repeatability and stability.