优化HPLC测定黑皮鸡枞菌中氨基酸含量样品处理的条件

Optimization Conditions of Processed Sample for the Determination of Amino Acid Content in Termitomyces spp. by HPLC

针对不同种类氨基酸采取不同水解方式,优化黑皮鸡枞菌的水解方法,利于高效液相色谱法(high performance liquid chromatography,HPLC)测定样品不同部位氨基酸的组成及含量。通过对黑皮鸡枞菌进行酸水解、碱水解和过甲酸氧化法处理,并进行正交试验优化酸水解的条件,以2,4-二硝基氟苯(2,4-dinitrofluorobenzene,DNFB)为柱前衍生剂,反相高效液相色谱法检测黑皮鸡枞菌菌伞、菌柄及子实体中的氨基酸组成。结果表明:当HCl浓度为6 mol/L、加量4 mL、110℃水解24 h时可获得较优的水解效果。采用HPLC检测两个地区产的黑皮鸡枞菌子实体和菌伞、菌柄中,3种水解方法结合处理的水解液,均检出21种氨基酸,总量分别为21.51%、23.68%;菌伞比菌柄中氨基酸含量约高1.28倍。3种水解方法结合处理样品可更全面的检测黑皮鸡枞菌中的氨基酸。

Different hydrolysis methods for different kinds of amino acids were used and optimised in Termitomyces spp.and that was better for the determination of the composition and content of amino acids in different parts of the sample by high performance liquid chromatography(HPLC).Acid hydrolysis,alkali hydrolysis and peroxy formic acid treatments were used in this study.And the orthogonal test was carried out to optimize the acid hydrolysis.With 2,4-dinitrofluorobenzene(DNFB)as the pre-column derivatization reagent,reversed phase HPLC was used for determination the content and composition of amino acids in pileus,stipe and sporophore of Termitomyces spp..The results showed that the optimal hydrolysis was obtained when the concentration of HCl was 6 mol/L,the adding dosage was 4 mL and hydrolysis at 110℃for 24 h.Amino acids in pileus,stipe and sporophore of Termitomyces spp.in two growing area was detected by HPLC combined with the three hydrolysis methods,and 21 kinds of amino acids were detected with 21.51%and 23.68%of the total,respectively.In the pileus,the amino acid content was about 1.28 times higher than that of stipe.Combined the sample with the three hydrolysis methods can be more comprehensive detection of the amino acids in Termitomyces spp.