摘要
目的:采用柱前衍生化法测定板蓝根颗粒中6种游离氨基酸的含量。方法:采用6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)进行柱前衍生化,超高效液相色谱法测定板蓝根颗粒中精氨酸、脯氨酸、苏氨酸、丙氨酸、γ-氨基丁酸、缬氨酸的含量。Waters AccQ Tag TM Ultra C18色谱柱(2.1 mm×100 mm,5μm),流动相AccQ Tag Ultra Eluent A与AccQ Tag Ul-tra Eluent B进行梯度洗脱,流速0.7 mL.min-1,柱温55℃,检测波长260 nm。结果:精氨酸的线性范围为4.155～49.86μg,r=0.999 9;苏氨酸的线性范围为0.595～5.95μg,r=0.999 8;丙氨酸的线性范围为0.445～4.45μg,r=0.999 9;γ-氨基丁酸的线性范围为0.515～5.15μg,r=0.999 9;脯氨酸的线性范围为8.858～106.3μg,r=0.999 9;缬氨酸的线性范围为0.585～5.85μg,r=0.999 8。平均回收率分别为100.6%,98.34%,98.35%,100.2%,98.44%,98.18%;平均RSD分别为1.8%,1.5%,1.9%,2.0%,2.4%,2.0%(n=6)。5家企业的的板蓝根颗粒中氨基酸类成分的含量存在差异。结论:该方法操作简单,重复性好,灵敏度高,结果准确可靠。
Objective： To establish a quantitative method with precolumn derivatization for determining the contents of six common amino acids in Banlangen Keli by UPLC. Method： Using 6-acetamido-4-hydroxy-2-methyl quinoline as the derivating agent, we determined the contents of arginine, threonine, alanine, gamma aminobutyric acid, proline, and valine. The UPLC analysis was performed on a Waters AccQ Tag TM Ultra C18 column （2.1 mm×100 mm, 5 μm） with mobile phase AccQ Tag Ultra Eluent A and AccQ Tag Ultra Eluent B gradient elution at a flow rate of 0.7 mL·min^-1. The column temperature was 55 ℃ and detection wavelength was 260 nm. Result： The linear ranges of arginine, thremine, alanine, gamma aminobutyric acid, proline, and valine were 4.155-49.86 μg （r=0.999 9）, 0.595-5.95 μg （r=0.999 8）, 0.445-4.45 μg （r=0.999 9）, 0.515-5.15 μg （r=0.999 9）, 8.858-106.3 μg （r=0.999 9）, 0.585-5.85 μg （r=0.999 8）.Their average recoveries were 100.6%, 98.35%, 100.2%, 98.44%, 98.34%, 98.18% with RSD 1.8%,1.9%, 2.0%, 2.4%, 1.5% and 2.0%, respectively （n=6）.The contents of amino acids were different in samples from five productive enterprises. Conclusion： The method is efficient, good reproducible, sensitive, and accurate.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2011年第23期3306-3309,共4页
China Journal of Chinese Materia Medica
基金
国家"十一五"科技支撑计划项目(2006BAI14B01)
