高效液相色谱—荧光法测定血清苯丙氨酸和酪氨酸  

Determination of phenylalanine and tyrosine in serum by high-performance liquid chromatography with fluorescent detection

作　　者：洪敏[1] 唐爱国[1] 

机构地区：[1]中南大学湘雅二医院检验科,长沙410011

出　　处：《中南大学学报（医学版）》2004年第1期67-71,共5页Journal of Central South University ：Medical Science

摘　　要：目的 :建立高效液相色谱 荧光法测定血清苯丙氨酸 (Phe)和酪氨酸 (Tyr)含量。方法 :血清标本经高氯酸溶液去蛋白质后 ,不需要荧光衍生 ,取上清液 2 0 μl直接进样 ,用HypersilC8色谱柱 (6 .0mm× 30 0mm ,10 μm)分离 ,流动相为乙腈 水 (体积比 10 :90 ) ,流速 1.0ml/min ,在激发光波长 2 10nm ,发射光波长 30 3nm的条件下定量测定Phe和Tyr。结果 :Phe线性范围为 12 .2 12 2 0 .0 μmol/L ,最低检测限为 3μmol/L ,批内变异系数 (CV)为4 .0 7% ,批间CV为 7.17% ,平均回收率 10 2 .5 % ;Tyr线性范围为 5 .5～ 5 5 0 .0 μmol/L ,最低检测限为 0 .8μmol/L ,批内CV为2 .99% ,批间CV为 5 .2 3% ,平均回收率 10 2 .2 %。色氨酸、5 羟色胺、5 羟吲哚乙酸等物质不干扰实验。整个分析过程可在 30min内完成 ;测定结果与HPLC 紫外法比较 ,两者有很好的相关性。结论 :HPLC 荧光法能同时测定血清Phe和Tyr浓度 ,且简便、快速。Objective To determine phenylalanine and tyrosine in serum by high-performance liquid chromatography (HPLC) method.Methods Serum precipitated with perchloric acid was isocratically eluted using a base-deactivated C8 column with 10% acetonitrile in water as the mobile phase. The flow rate was 1.0 ml/min. Fluorescent detector worked at an excitation wavelength of 210 nm and emission wavelength of 303 nm. Results Good linearity was observed in the range of 12.2~1 220.0 μmol/L for phenylalanine, the intra- and inter-assay coefficients of variation (CV) were 4.07% and 7.17% respectively, and the average recovery was 102.5%. There also was a good linearity in the range of 5.5~ 550.0 μmol/L for tyrosine. The intra- and inter-assay coefficients of variation were 2.99% and 5.23% respectively, and the average recovery was 102.2%. The total assay time for the serum sample was within 30 min.We found a good correlation compared with HPLC ultraviolet detection.Conclusion This method is simple, rapid, accurate in measuring phenylalanine and tyrosine in serum quantitatively.

关 键 词：高效液相色谱-荧光法 血清学测定 苯丙氨酸 酪氨酸 精密度 回收率 

分 类 号：O657.72[理学—分析化学]