【摘要】 目的:建立测定野生四叶参中氨基酸含量的反相高效液相色谱法(RP-HPLC)。方法:用盐酸水解大别山区野生四叶参中的蛋白质。采用反相高效液相色谱法,色谱条件为:色谱柱:HypersilODS-BP C18柱(4.6mm×250mm,5μm);流动相A:醋酸-醋酸钠缓冲液(pH=7.2);流动相B:甲醇,流速1.0mL/min,流动相采用梯度洗脱方式,荧光检测波长为450nm。分析四叶参中各种氨基酸含量。采用外标法定量。结果:氨基酸在25.0~800.0μmol/L范围内浓度与色谱峰面积有良好的线性关系(r≥0.993),该方法的回收率在89.5%~109.2%,RSD在3.2%~9.0%,检测限在1.0~3.7μmol/L。野生四叶参中含有12种氨基酸,总氨基酸含量为51.49mg/g,必需氨基酸含量为6.95mg/g。结论:建立的分析方法精密度好、准确度高,适用于野生四叶参中氨基酸含量的分析。更多还原

【Abstract】 Aim:To determine the contents of amino acids in Wild Codonopis Lanceolata by RP-HPLC.Methods:Proteins were hydrolyzed by 6 mol/L HCl. The separation was performed in a Hypersil ODS-BP C18 (4.6 mm×250 mm,5 μm)column with a mobile phase of NaAc-HAc-CH3OH(gradient elution).The flow rate was 1.0 mL/min. RP-HPLC was applied to determine the contents of amino acids at 450 nm with fluorescence detection(FLD)with OPA pre-column residuederivation.Results: The calibration curves were linear with 25.0～800.0 μmoL/L between the peak area and concentration,and correlation coefficient was higher than 0.993.RSD was 3.2%～9.0%,and the recovery rate was 89.5%~109.2%. There were 12 kinds of amino acids,the total content of amino acids was 51.49 mg/g,and the content of essential amino acids was 6.95 mg/g.Conclusion: This method is simple, rapid, sensitive, and accurate, and can be used for the determination of amino acids in Wild Codonopis Lanceolata.更多还原