建立了醋酸锌在线衍生高效液相色谱法同时测定血浆中色氨酸(Trp)、犬尿氨酸(Kyn)、5-羟吲哚乙酸(5-H iaa)和犬尿喹啉酸(Kyna)的方法.以3-硝基酪氨酸为内标(IS),采用Hypersil C-18柱(250 mm×4.0 mm,5μm),以250 mmol/L醋酸锌溶液(pH 5.5)-乙腈(95∶5,v/v)为流动相,流速为0.8 mL/min,柱温30℃.荧光检测波长设定:5-Hiaa为278 nm(λex)/343 nm(λem),Kyna为244 nm(λex)/400 nm(λem);紫外检测波长设定:Kyn和IS为360 nm,Trp为302 nm.4种物质的回收率在91.62%~114.17%之间;线性范围分别为2.50 ~ 320.00μmol/L(Trp),0.32~15.36 μmol/L(Kyn),3.27 ~ 104.60 nmol/L(5-Hiaa),14.00 ~ 464.80 nmol/L(Kyna);检出限分别为0.078 μmol/L(Trp),0.056 μmol/L(Kyn),0.690 nmol/L(5-Hiaa),1.290 nmol/L(Kyna).利用该方法对30例正常孕妇和28例女性健康志愿者的血浆进行测定,结果表明两组间Trp,Kyn和Kyna含量有显著性差异.该方法操作简便,重复性好,灵敏度高,适合于临床检测.
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