柱前衍生化HPLC测定中药材氨基酸的含量

目的:建立中药材有效成份氨基酸柱前衍生HPLC分析测定方法。方法:从常见中药材党参和山楂中提取氨基酸,邻苯二甲醛(OPA)柱前衍生,反相高效液相色谱分离测定,以保留时间定性,面积外标法定量。结果:水解温度108℃,提取24 h,可有效提取药材中的氨基酸;梯度洗脱程序为(t,C%):(t:保留时间C:甲醇):(0,0);(9,30);(16,45);(25,60);(28,65);(32,80),二种药材中的十六种氨基酸得到较好分离和定性定量结果,其中天冬氨酸和胱氨酸的平均加样回收率分别为101.0%和94.4%。结论:采用HPLC柱前衍生分离分析党参和山楂中十六种氨基酸的方法灵敏、简便、快捷、高效,适于推广应用,可为解决中药材氨基酸分离测定难题提供参考。

Determination of Amino Acids in Chinese Medicinal Materials by HPLC with Pre-column Derivation

Objective To establish an effective method by which we can determine the content and concentration of amino acids in them.Methods The most suitable procedures are selected and applied to the extraction and further purification of amino acids from common herbs;Employing OPA as a pre-column derivative reagent,reversed-phase(RP) high performance liquid chromatogram(HPLC) is applied to detect.Both qualitative and quantitative analyses are separa-tely performed on sample amino acids recording to retentive time and peak area(with exterior standard).Results It is advisable to extract amino acid under the proper conditions of hydrolyzing temperature at 108 degree and hydrolyzing duration of 24 hours.When gradient elution of flowing phase is taken as(t,C%):(0,0);(9,30);(16,45);(25,60);(28,65);(32,80),where it is retentive time and C is methanol amount;sixteen amino acids can be obtained from Radix Codonpsispilosulae and hawthorn respectively.The average recoveries are 101.0% and 94.4% separately for Cystine and Aspartic acid therein.Conclusion It is reco-mmendable to make analysis on amino acids by pre-column derivatization method,which is inventive,effective,simple and convenient.It can supply a reference for dealing with the difficulties in analysis of amino acids.