|
For effective determination of methylthiohydantoin amino acids(MTHs) by gas liquid chromatography in the protein sequencing, derivatization with N-methyl-N-(tert.-butyl-dimethylsilyl)trifluoroacetamide(MTBSTFA), a new silylating reagents, was attempted instead of trimethylsilyl(TMSi) derivatives by N,O-bis(trimethylsilyl)trifluoroacetamide(BSTFA) used up to the present and N(O)-butyldimethylsilyl MTHs derivatized by MTBSTFA were analysed on HP-1 capillary column. Twenty one protein amino acids except cystine were indentified. Especially arginine that did not detected with TMSi derivative on packed column until now was resolved by derivatization with MTBSTFA. N(O)-butyldimethylsilyl MTHs showed multiple peaks by MTBSTFA were proline, isoleucine, glycine and tyrosine and hydroxyproline especially showed several extraneous peaks more than two. Calibration curves of N(O)-butyldimethysilyl MTHs of amino acids in the range of 2.5 nmol∼7.5 nmole showed good linearity. however, those of lysine, histidine and arginine showed linearity in the range of 5.0 nmole∼15.0 nmole. Correlation coefficients and regression coefficients of all calibration curves were highly significant(p$lt;0.001)(Received February 25, 1992, accepted April 17, 1992).
|