Semi-automated solid-phase extraction method for studying the biodegradation of ochratoxin A by human intestinal microbiota

详细信息    查看全文

• 作者：Valé ; rie Camel^a ; ^b ; ^{valerie.camel@agroparistech.fr} ; Minale Ouethrani^a ; ^b ; Cindy Coudray^a ; ^b ; Catherine Philippe^c ; ^d ; Sylvie Rabot^c ; ^d
• 关键词：Biodegradation ; Human intestinal microbiota ; Ochratoxin A ; Solid-phase extraction
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2012
• 出版时间：15 April, 2012
• 年：2012
• 卷：893-894
• 期：Complete
• 页码：63-68
• 全文大小：456 K

文摘

A simple and rapid semi-automated solid-phase (SPE) extraction method has been developed for the analysis of ochratoxin A in aqueous matrices related to biodegradation experiments (namely digestive contents and faecal excreta), with a view of using this method to follow OTA biodegradation by human intestinal microbiota. Influence of extraction parameters that could affect semi-automated SPE efficiency was studied, using C18-silica as the sorbent and water as the simplest matrix, being further applied to the matrices of interest. Conditions finally retained were as follows: 5-mL aqueous samples (pH 3) containing an organic modifier (20 % ACN) were applied on 100-mg cartridges. After drying (9 mL of air), the cartridge was rinsed with 5-mL H₂O/ACN (80:20, v/v), before eluting the compounds with 3¡Á 1 mL of MeOH/THF (10:90, v/v). Acceptable recoveries and limits of quantification could be obtained considering the complexity of the investigated matrices and the low volumes sampled; this method was also suitable for the analysis of ochratoxin B in faecal extracts. Applicability of the method is illustrated by preliminary results of ochratoxin A biodegradation studies by human intestinal microbiota under simple in vitro conditions. Interestingly, partial degradation of ochratoxin A was observed, with efficiencies ranging from 14 % to 47 % after 72 h incubation. In addition, three phase I metabolites could be identified using high resolution mass spectrometry, namely ochratoxin ¦Á, open ochratoxin A and ochratoxin B.