高效液相色谱-荧光检测法测定血清色氨酸
[Determination of serum tryptophan by high performance liquid chromatography with fluorescence detection].
作者信息
Tang Ai-Guo, Wang Qing-Ping
机构信息
Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
出版信息
Hunan Yi Ke Da Xue Xue Bao. 2002 Dec 28;27(6):569-71.
OBJECTIVE
To develop a rapid assay for determining tryptophan in human serum.
METHODS
The serum sample was injected into the high performance liquid chromatography (HPLC) system after the precipitation of protein with perchloric acid, and was monitored with a fluorometric detector with wavelength settings of 254 nm for excitation and 313 nm for emission. The mobile phase is 20% acetonitrile.
RESULTS
Good linearity was observed in 1.22-400 mumol.L-1 range for tryptophan. The detection limit was 0.1 mumol.L-1. The intra- and inter-assay coefficients of variation (CV) were 2.57% and 3.66%, respectively. The average recovery was 97.13%. The detection time of serum tryptophan was within 5 min. Phenyalanine, tyrosine, serotonin, and 5-hydroxyindoleacetic acid didn't interfere in the assay.
CONCLUSION
This method is simple, rapid, sensitive, accurate, and suitable for clinical application.
目的
建立一种快速检测人血清中色氨酸的方法。
方法
血清样品用高氯酸沉淀蛋白质后注入高效液相色谱（HPLC）系统，用荧光检测器监测，激发波长设置为254nm，发射波长设置为313nm。流动相为20%乙腈。
结果
色氨酸在1.22 - 400μmol·L⁻¹范围内呈良好线性。检测限为0.1μmol·L⁻¹。批内和批间变异系数（CV）分别为2.57%和3.66%。平均回收率为97.13%。血清色氨酸的检测时间在5分钟内。苯丙氨酸、酪氨酸、血清素和5-羟吲哚乙酸不干扰该测定。
结论
该方法简单、快速、灵敏、准确，适用于临床应用。
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