More rapid and sensitive method for simultaneous determination of tryptophan and kynurenic acid by HPLC.
OBJECTIVE
To describe a simple, rapid, and sensitive HPLC method for simultaneous determination of TRP and KYNA in human serum.


METHODS
Samples were deproteinized by perchloric acid. KYNA was detected at 344 nm of excitation wavelength and 404 nm of emission wavelength, TRP was detected at 254 nm and 404 nm, with a total run time of 13 min per sample.


RESULTS
Standard curves of 0.49 micromol/L to 196 micromol/L of TRP were linear. Inter-day and intra-day coefficient of variations were 3.31% and 4.14%, respectively. Average recovery was 104.43%. Detection limit was 0.001 micromol/L. The linearity of the assay was maintained from 1.5 nmol/L to 2093 nmol/L of KYNA. Inter-day and intra-day CVS were 3.20% and 4.27%, respectively. Average recovery was 101.19%. Detection limit was 0.05 nmol/L.


CONCLUSIONS
The developed HPLC method is simple, convenient and can be applied in the diagnosis of related diseases.
Pi
Lan-gan
LG
Department of Clinical Laboratory, The Second Xiangya Hospital, Central South University, No.139, Renmingzhong Road, Changsha, Hunan 410011, China.
Tang
Ai-guo
AG
Mo
Xi-ming
XM
Luo
Xi-bo
XB
Ao
Xiang
X
D000097:Acetonitriles / Q000737:chemistry; D000293:Adolescent; D000328:Adult; D044466:Asian People; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006262:Health; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007736:Kynurenic Acid / Q000097:blood*; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D012015:Reference Standards; D012044:Regression Analysis; D015203:Reproducibility of Results; D012212:Rheology; D012680:Sensitivity and Specificity; D013997:Time Factors; D014364:Tryptophan / Q000097:blood*; D055815:Young Adult; D019345:Zinc Acetate / Q000737:chemistry
D000097:Acetonitriles; D014364:Tryptophan; D019345:Zinc Acetate; D007736:Kynurenic Acid; C032159:acetonitrile