反相HPLC法测定家兔血清中N-乙酰色氨酸及其代谢产物

Reverse phase HPLC analysis of N-ATRP and its metabolite TRP in rabbit serum

以ODS C18柱为固定相,0.1mol·L^(-1)NaAc-25%甲醇水为流动相,分离家兔血清中的N-乙酰L-色氨酸(N-ATRP)及其代谢产物L-色氨酸(TRP),用荧光检测器(激发光280nm,发射光360nm)检测其天然荧光.血清中物质对该方法干扰较少,线性范围在1.0～17.0mg·L^(-1)之间,N-ATRP和TRP的回收率分别为101.8%±6.5%和92.2%±5.2%精确度和重现性较好,变异系数均在10%以内,测得的结果表明N-ATRP(27mg·kg^(-1))给予家兔一次静注后在血中迅速分解为TRP,T_(1/2)为8.5min.

A simple, rapid and sensitive method for reversed phase HPLC determination of N-acetyl-L-tryptophan (N-ATRP) and its metabolite L-tryptophan (TRP) in rabbit serum was developed based on their intrinsic fluorescence using excitation and emission wavelengths of 280 and 360 nm, respectively. The separation of two amino acids was achieved on an ODS C18 column with a mobile phase of 0. 1 mol ?L-1 NaAc-25% methanol (pH 5. 7) at a flow rate of 0. 5 ml ?min-1. The calibration curves for N-A-TRP and TRP provided the linear range about1.0 - 17. 0mg ?L-1(r=0. 9962 and 0.9978, respectively). The recoveries greater than 98% and good reproducibility were achieved with rabbit serum. The concentration-time curve of N-ATRP after iv administration was determined in rabbits. Results showed that N-ATRP was quickly converted to TRP with a half-life of 8. 5 min. The decrese of N-ATRP accorded well with the rise of TRP.