超高效液相色谱-荧光法测定大鼠尿液中的黄蝶呤、异黄蝶呤、酪氨酸和色氨酸

Determination of xanthopterin,isoxanthopterin,tyrosine and tryptophan in mouse urine by UPLC-LD

建立了一种大鼠尿液中黄蝶呤、异黄蝶呤、酪氨酸和色氨酸的超高效液相色谱-荧光分析方法(UPLCFLD),大鼠尿液利用甲醇和乙腈除蛋白后,采取C18固相萃取小柱进行净化,然后用UPLC-FLD法进行测定。黄蝶呤、异黄蝶呤、酪氨酸和色氨酸分别在0.01~10.0,0.005~10.0,0.5~100,0.5~200mg·L-1范围内与色谱峰面积呈线性关系,4种物质的仪器检测限分别为0.005,0.000 3,0.004和0.002mg·L-1,加标回收率除黄蝶呤在61.1%~78.0%之间,其余各物质回收率均在86.0%~110.0%之间,RSD小于10.5%。

A simple,rapid and sensitive method for the determination of xanthopterin,isoxanthopterin,tyrosine and tryptophan in mouse urine by UPLC-FLD was proposed in this study.After deproteinized with methanol and acetonitrile,the mouse urine samples were purified with C18solid phase extraction column and then detected directly by UPLC-FLD.Xanthopterin,isoxanthopterin,tyrosine and tryptophan behaved linearly in the range of 0.01~10.0mg/L,0.005~10.0mg·L-1,0.5~100mg·L-1 and 0.5~200mg·L-1,respectively.The limits of instrument detection of the four analytes were found to be 0.005、0.0003、0.004and 0.002mg·L-1,respectively.The recoveries were in the range of 86.0% ~110.0%except that of xanthopterin were in the range of 61.1% ~78.0%.The relative standard deviations were lower than10.5%.