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[Determination of serum tryptophan by high performance liquid chromatography with fluorescence detection]
1Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
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.