[Rapid analysis of tryptophan in serum by high performance liquid chromatography with fluorescence detection].
A sensitive, rapid and specific method for the determination of tryptophan(Trp) in serum by high performance liquid chromatography with fluorescence detection (HPLC-FLD) has been established. Serum sample was precipitated by 5% perchloric acid solution and centrifuged to remove protein and then assayed by HPLC-FLD. The operating conditions were Nova-Pak C18 column (3.9 mm i.d x 150 mm, 4 microns), 5 mmol/L KH2PO4 as the mobile phase at a flow rate of 1.0 mL/min. The fluorescence detector was operated at lambda ex 254 nm and lambda em 338 nm. The method was proved to be linear in the range of 0.49 mumol/L-490.00 mumol/L with a regression coefficient of 0.9999. The minimum detection limit was 0.10 mumol/L. The recoveries were 97.2%-98.6%. The intra-batch and inter-batch RSDs were 2.0% and 3.2% respectively. The detection time of serum Trp was within 5 min after injection. The results suggest that this method is simple, fast, accurate and convenient.
Wang
Qing-ping
QP
Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China. wangyulin774@sohu.com
Tang
Ai-guo
AG
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000097:blood*
D014364:Tryptophan