Quantification of microbial uptake of quercetin and its derivatives using an UHPLC-ESI-QTOF mass spectrometry assay

Abstract

A liquid chromatography-mass spectrometry-based method for the quantification of intracellular concentrations of dietary phenolics in bacteria was developed. Using this assay, the time-dependent uptake of quercetin and two of its glycosides into gut microbiota model organisms (Escherichia coli and Bifidobacterium bifidum) under aerobic and anaerobic conditions was studied. For the first time, quantitative data on the uptake of dietary phenols were obtained indicating a time-dependent differential uptake of the dietary compounds. The shape of the uptake curve and the comparative use of bacterial mutants lacking porins OmpFC or the multi-drug efflux pump AcrAB, respectively, along with the identification and quantification of selected bacterial metabolites provided a mechanistic insight into the uptake process.

Role of the small intestine, colon and microbiota in determining the metabolic fate of polyphenols

“…Under anaerobic conditions, E. coli mutants lacking either porins 0mpF and 0mpC or the efflux transporter multi-drug efflux pump AcrAB, did not differ in behaviour relative to the wild type [132], and so the precise uptake mechanisms remain unclear. were applied to the in vitro catabolism of black tea or a mixture of red wine and grape juice considerable variation was observed, each individual's microbiota producing a distinct catabolite profile [89].…”

“…Under anaerobic conditions, E. coli mutants lacking either porins 0mpF and 0mpC or the efflux transporter multi-drug efflux pump AcrAB, did not differ in behaviour relative to the wild type [132], and so the precise uptake mechanisms remain unclear. bulgaricus grew best in the presence of oligomeric procyanidins.…”

Pharma‐molecule Transport across Bacterial Membranes: Detection and Quantification Approaches by Electrochemistry and Bioanalytical Methods

“…Chromatographic methods serve to separate the components of a sample on the basis of intrinsic properties, such as mass, polarity, or charge. Chromatographic methods are frequently employed in bacterial transport studies [7–10, 12, 16, 21, 26, 27, 30, 33–35, 72, 77, 84–86] and techniques include high‐performance liquid chromatography (HPLC) [8, 21, 33, 34] and liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) [10, 26, 35, 85] …”

“…Depending on the method, quantitative transport properties can vary. For example, several papers retrieved by the review present research regarding the expulsion of materials via the AcrAB‐TolC efflux pump [6, 19, 26, 27, 47, 48, 74, 84] . The methods specified in these papers include spectrofluorimetry, [6, 74] LC‐MS/MS, [26] and a hydrolysis‐efflux coupling assay [19, 48] .…”

Ácidos clorogénicos presentes en el café: capacidad antimicrobiana y antioxidante

“…El café posee importantes cantidades de compuestos fenólicos cuyos representantes más abundantes son los ácidos clorogénicos; estos compuestos poseen diferentes actividades biológicas ya descritas, sin embargo, se ha reportado que los efectos atribuidos al consumo de ácidos clorogénicos no son necesariamente consecuencia de su capacidad antioxidante, ya que, en su transitar por el sistema gastrointestinal son catabolizados a una serie de metabolitos que son los que al final van a predominar en el sistema circulatorio y llevar a cabo las diferentes acciones que se traducen en beneficios asociados al consumo de café (Ludwig et al, 2013;Hakeem et al, 2016;. Nakayama y Oishi (2013), mediante pruebas in vivo con el empleo de ratones, reportaron que el consumo de café puede ayudar a regular la microbiota intestinal y, por lo tanto, contribuye al mantenimiento del balance del mismo.…”