一测多评法测定羊肚菌中17种氨基酸的含量

目的:建立以谷氨酸为内参物，同时测定羊肚菌中17种氨基酸含量的一测多评法。方法:采用高效液相色谱，以乙腈-乙酸铵水溶液为流动相，梯度洗脱，以谷氨酸为内参物，建立其与其他16种氨基酸的相对校正因子（RCF），并用相对校正因子计算16种氨基酸的含量；同时采用内标法对羊肚菌中17种氨基酸进行测定，比较一测多评法与内标法结果的差异，并对方法耐用性进行考察。结果:17种氨基酸在线性范围内线性关系良好，线性相关系数R2>0.9990，平均加标回收率在92.42%~101.16%之间，RSD在0.25%~1.94%，在不同实验条件下重现性良好（RSD<2.0%）；一测多评法和内标法测定结果的对比分析，无显著性差异（P>0.05）。在不同色谱柱、色谱体系中的相对校正因子、相对保留时间的RSD均小于2.0%，耐用性良好。结论:本文建立的一测多评法简单易操作，结果准确，节约了测定成本，为羊肚菌中氨基酸的测定提供了一种新的模式，为羊肚菌的质量评价提供有价值的参考。

Determination of Seventeen Amino Acids in Morchella Esculenta by Quantitative Analysis of Multi-Components by Single-Marker Method

Objective: To establish a quantitative analysis of multi-components by single-marker (QAMS) with using glutamic acid as the internal reference, the content of 17 amino acids in Morchella esculenta was determined simultaneously. Methods: High performance liquid chromatography (HPLC) was used with acetonitrile-ammonium acetate aqueous solution as mobile phase, gradient elution and glutamic as the internal reference substance was used to establish its relative correction factors (RCF) of other 16 amino acids. The content of 17 amino acids in Morchella esculenta was calculated by using the relative correction factor. Internal standard method was used to determine 17 amino acids in Morchella esculenta. The validity of the QAMS method was evaluated by comparison of the quantitative results of both methods and to validate the durability of this method. Results: The 17 amino acids had a great linear relationship in the linear range, and the correlation coefficients higher than 0.9990, the average recovery rate of 17 amino acids were in the range of 92.42%~101.16%, and the relative standard deviations (RSD) were in the range of 0.25%~1.94%, and reproducibility was good in different experimental conditions (RSD<2.0%); through the comparative analysis of determination results of QAMS and the internal standanrd method, the two methods had no significant difference (P>0.05). The RCF and the relative retention time RSD in different chromatographic columns and chromatographic systems all less than 2.0%, and the durability was good. Conclusion: This method is simple, easy to operate, accurate and can save the cost of content determination. It can be used as a new model for the determination of amino acid content in Morchella esculenta, and can also provide a valuable reference for the quality evaluation of Morchella esculenta.