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目的:建立同时测定人血清中的犬尿氨酸和色氨酸浓度的方法。方法:血清样品用24%高氯酸以10:1进行蛋白沉淀处理,取上清液进样分析。采用高效液相色谱-紫外/荧光检测器法分别检测犬尿氨酸和色氨酸浓度,色谱柱为Accurasil-C18,流动相为乙酸钠-乙腈溶液(24.8mmol.L-1乙酸钠,用乙酸调pH值至约4.0)=90:10,流速1.0mL.min-1,紫外检测波长为360nm,荧光检测激发波长为285nm、发射波长为365nm。结果:犬尿氨酸、色氨酸血药浓度分别在0.16～20.89(r=0.9999)、0.24～61.58μmol.L-1(r=0.9996)范围线性关系良好;高、中、低3种浓度的方法回收率分别为91.33%～101.82%、91.49%～104.37%;日内及日间RSD均<6%。结论:本方法稳定、可行,可为临床疾病的诊断和药物治疗提供实验室依据。
Objective: To establish a method for simultaneous determination of kynurenine and tryptophan in human serum. Methods: Serum samples were treated with 24% perchloric acid at 10: 1 for protein precipitation and the supernatant was injected for analysis. The concentrations of kynurenine and tryptophan were determined by HPLC-UV / fluorescence detector. The chromatographic column was Accurasil-C18, and the mobile phase was sodium acetate-acetonitrile solution (24.8 mmol.L-1 sodium acetate, Acetic acid to adjust the pH to about 4.0) = 90: 10, flow rate 1.0mL.min-1, UV detection wavelength of 360nm, fluorescence detection excitation wavelength of 285nm, emission wavelength of 365nm. Results: The concentrations of kynurenine and tryptophan were linear in the range of 0.16-20.89 (r = 0.9999) and 0.24-61.58μmol.L-1 (r = 0.9996), respectively. The concentrations of kynurenine, tryptophan, The recoveries were 91.33% ~ 101.82% and 91.49% ~ 104.37%, respectively. The intra-day and inter-day RSDs were all less than 6%. Conclusion: The method is stable and feasible, which can provide laboratory evidence for clinical diagnosis and drug therapy.