Structure-activity relationships for the fluorescence of ochratoxin A: insight for detection of ochratoxin A metabolites.
Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium that is widely found as a contaminant of food products. The toxin is a renal carcinogen in male rats, the cause of mycotoxicoses in pigs and has been associated with chronic human kidney diseases. Bioactivation has been implicated in OTA-mediated toxicity, although inconsistent results have been reported, due, in part, to the difficulty in detecting OTA metabolites in vivo. Liquid chromatography (LC) coupled with fluorescence detection (FLD) is the most widely used analytical detection method for OTA. Under acidic conditions the toxin generates blue fluorescence (465 nm) that is due to an excited state intramolecular proton transfer (ESIPT) process that generates an emissive keto tautomer. Disruption of this ESIPT process quenches fluorescence intensity and causes a blue shift in emission maxima. The aim of the present study was to determine the impact of the C5-chlorine atom, the lactone moiety and the amide bond on OTA fluorescence and derive optical parameters for OTA metabolites that have been detected in vitro. Our results highlight the limitations of LC/FLD for OTA metabolites that do not undergo ESIPT. For emissive derivatives, our absorption and emission data improves the sensitivity of LC/FLD (3-4-fold increase in the limit of detection (LOD)) for OTA analogues bearing a C5-OH group, such as the hydroquinone (OTHQ) metabolite and the glutathione conjugate of OTA (OTA-GSH). This increased sensitivity may facilitate the detection of OTA metabolites bearing a C5-OH group in biological fluids and enhance our understanding of OTA-mediated toxicity.
Frenette
Christine
C
Department of Chemistry, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Paugh
Robert J
RJ
Tozlovanu
Mariana
M
Juzio
Maud
M
Pfohl-Leszkowicz
Annie
A
Manderville
Richard A
RA
D000577:Amides / Q000737:chemistry; D002244:Carbon / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D007783:Lactones / Q000737:chemistry; D015394:Molecular Structure; D009705:Nucleosides / Q000737:chemistry; D009793:Ochratoxins / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D010777:Photochemistry; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D000577:Amides; D007783:Lactones; D009705:Nucleosides; D009793:Ochratoxins; C025589:ochratoxin A; D002244:Carbon