Identification and in vitro cytotoxicity of ochratoxin A degradation products formed during coffee roasting.
The mycotoxin ochratoxin A is degraded by up to 90% during coffee roasting. In order to investigate this degradation, model heating experiments with ochratoxin A were carried out, and the reaction products were analyzed by HPLC-DAD and HPLC-MS/MS. Two ochratoxin A degradation products were identified, and their structure and absolute configuration were determined. As degradation reactions, the isomerization to 14-(R)-ochratoxin A and the decarboxylation to 14-decarboxy-ochratoxin A were identified. Subsequently, an analytical method for the determination of these compounds in roasted coffee was developed. Quantification was carried out by HPLC-MS/MS and the use of stable isotope dilution analysis. By using this method for the analysis of 15 coffee samples from the German market, it could be shown that, during coffee roasting, the ochratoxin A diastereomer 14-(R)-ochratoxin A was formed in amounts of up to 25.6% relative to ochratoxin A. The decarboxylation product was formed only in traces. For toxicity evaluations, first preliminary cell culture assays were performed with the two new substances. Both degradation products exhibited higher IC50 values and caused apoptotic effects with higher concentrations than ochratoxin A in cultured human kidney epithelial cells. Thus, these cell culture data suggest that the degradation products are less cytotoxic than ochratoxin A.
Cramer
Benedikt
B
Institut für Lebensmittelchemie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 45, 48149 Münster, Germany.
Königs
Maika
M
Humpf
Hans-Ulrich
HU
D002461:Cell Line, Transformed; D002851:Chromatography, High Pressure Liquid; D040503:Coffea / Q000737:chemistry*; D004847:Epithelial Cells; D005506:Food Contamination / Q000517:prevention & control; D005511:Food Handling / Q000379:methods*; D006358:Hot Temperature; D006801:Humans; D007668:Kidney; D009793:Ochratoxins / Q000032:analysis / Q000737:chemistry* / Q000633:toxicity; D012639:Seeds / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
D009793:Ochratoxins; C025589:ochratoxin A