Simultaneous Determination of Contents of 17 Amino Acids in Fritillariae Pallidiflorae Bulbus Produced in Xinjiang by Post-column Derivatization Cation-exchange Chromatography / 中国药房

ABSTRACT

OBJECTIVE:To establish a method of contents determination of 17 amino acids in Fritillariae Pallidiflorae Bulbus produced in Xinjiang. METHODS:Cation-exchange column was used to separate 17 kinds of amino acids;post-column derivatiza-tion liquid chromatography was used to determine the contents of amino acidsthe column was strong sulfonic acid cation exchange resin LCAK06/Na with mobile phase A(weighing trisodium citrate 11.9 g,citric acid 6 g,phenol 1 g,dissolving by water,then adding 65 mL of methanol and 6 mL of concentrated hydrochloric acid,bringing the volume with tap water,adjusting pH to 3.4), mobile phase B(weighing trisodium citrate 11.9 g,NaOH 2.8 g,phenol 1 g,boric acid 5.0 g,adding water to dissolve,adjustingpH to 10.8),gradient elution,flow rate was 0.45 mL/min for A pump and 0.25 mL/min for M pump,the detection wavelength was 440 nm for proline and 570 nm for the remaining amino acids,the injection volume was 50 μL. RESULTS:The linear range were 20~400 nmol/mL of aspartic acid,threonine,serine,glutamic acid,glycine,alanine,valine,methionine,isoleucine,leucine,ty-rosine,phenylalanine,histidine,lysine,arginine,proline,10-200 nmol/mL of cystine(r were higher than 0.9890);RSDs of preci-sion,stability and reproducibility tests were lower than 2.0%;limits of detection were 0.16 nmol/mL except for cystine(0.08 nmol/mL);recovery was 98.5%-99.5%(RSD was 0.06%-0.21%,n=6). CONCLUSIONS:The method is simple with good precision, stability and reproducibility,and can be used for the simultaneous determination of amino acids in Fritillariae Pallidiflorae Bulbus produced in Xinjiang.