摘要
建立了反相高效液相色谱-蒸发光散射检测（HPLC-ELSD）直接测定6种未衍生游离氨基酸（脯氨酸Pro、缬氨酸Val、蛋氨酸Met、异亮氨酸Ile、亮氨酸Leu、苯丙氨酸Phe）含量的分析方法。采用VenusilMP-C18色谱柱,以甲醇和水为流动相进行线性梯度洗脱,流速为0.8mL·min-1,漂移管温度40℃,氮气流速2.5L·min-1;优化了流动相的洗脱条件。结果表明,在优化实验条件下,6种氨基酸峰面积的对数值与氨基酸质量浓度的对数值呈良好线性,检出限为10.0～15.0mg·mL-1,方法精密度及稳定性良好。采用该法对微波法提取的蛹虫草子实体中的氨基酸进行测定,待检蛹虫草样本中不含缬氨酸（Val）,其余5种氨基酸的总量为16.00mg/g,约占蛹虫草总质量的1.60%,其中脯氨酸（Pro）（占0.72%）和苯氨酸（Phe）（占0.56%）的含量较高。已检出的5种Pro、Met、Ile、Leu、Phe含量分别为7.20、0.88、0.69、1.65、5.58mg/g。
A high performance liquid chromatographic method coupled with evaporative light scattering detection（HPLC-ELSD）was developed for the determination of six underivatized free amino acids（Pro,Val,Met,Ile,Leu,Phe）.The sample was separated on a Venusil MP-C18 column using methanol-water as mobile phase by linear gradient elution.The optimal conditions were as the follows：flow rate：0.8 mL·min-1,drift tube temperature：40 ℃,nitrogen flow rate：2.5 L·min-1.The mobile phase elution conditions were optimized and the working system applicability was tested.The results showed that,under the optimal conditions,the logarithm correlation between peak area and amino acid concentration was in a good linearity.The detection limits of six amino acids were in the range of 10.0-15.0 mg·L-1.The method was applied in the detection of 6 amino acids in the fruiting body of Cordyceps militaris,and the results showed that except for Val,the other five amino acids（Pro,Met,Ile,Leu,Phe）were detected in the tested sample,and their total amount was 16.00 mg/g,accounting for 1.60% of the total mass of Cordyceps militaris,in which Pro（occupying 0.72% of the total mass of Cordyceps militaris）and Phe（occupying 0.56% of the total mass of Cordyceps militaris）were higher than the other three amino acids.The detected contents of five amino acids of Pro,Met,Ile,Leu and Phe were 7.20,0.88,0.69,1.65 and 5.58 mg/g,respectively.
出处
《分析测试学报》
CAS
CSCD
北大核心
2010年第10期1091-1094,共4页
Journal of Instrumental Analysis
基金
国家自然科学基金资助项目(30972287)
关键词
高效液相色谱
微波提取
蛹虫草
蒸发光散射检测器
high performance liquid chromatography
microwave extraction
Cordyceps militaris
evaporative light-scattering detector
