Abstract
BACKGROUND: The accumulation of quinolinic acid (QUIN) in cerebrospinal fluid and serum may be used as a biomarker for various neuropsychiatric and inflammatory diseases. In this study, we developed a highly sensitive method to measure QUIN. METHODS: A reverse-phase high-performance liquid chromatography (HPLC) with fluorescence detection was established based on the enzymatic conversion of QUIN to nicotinic acid mononucleotide by recombinant quinolinic acid phosphoribosyltransferase, followed by the formation of fluorescent (BODIPY)-labeled deamido-NAD by recombinant nicotinic acid mononucleotide adenyltransferase. RESULTS: BODIPY-deamido-NAD was isocratically eluted within 6 minutes using reverse-phase chromatography and its chromatographic peak was resolved. The calibration range, precision, and analytical recovery of the QUIN assay are suitable for the analysis of biological fluids. Compared with published quantitation limits for QUIN measurement by HPLC, this method is at least 30-fold more sensitive and has a lower limit of detection of 5.0 nmol/L. The sensitivity was comparable to that previously reported for gas chromatography/mass spectrometry (GC/MS) and the quantitation results of QUIN from samples of cerebrospinal fluid correlated well with that of the GC/MS method. CONCLUSIONS: We established a novel method to quantify QUIN in biological samples. Due to its high sensitivity and the fact that it does not rely on MS instrumentation, this method has the potential for widespread adoption in research laboratories.