Biosystems to detect chemical contaminants
AssigneeFundacion Azti-Azti Fundazioa
InventorsAlejandro BARRANCO IBARBIA, Amaia EREÑO ARTABE, Hugo Flávio CUNHA DA SILVA
Life of this patent
Estimating the term…
Abstract
The present invention refers to the field of food safety and environmental control. In particular, it refers to a method for the detection of chemical contaminants based on cytochrome P450 enzymes, to a kit to carry out said method and to its use.
Claims47
A method for the detection of a cytochrome P450 (CYP450) enzyme inhibitor chemical compound in a sample comprising the following steps:
- a) contacting the sample or an extract thereof with a CYP450 enzyme allowing the interaction between the chemical compound and the CYP450;
- b) addition of a CYP450 enzyme's substrate and incubation at suitable conditions to allow a catalytic reaction between the substrate and the CYP450, that generates a detectable signal;
- c) detection and measurement of the detectable signal;
- d) determination of the presence of the compound by comparing the measured detectable signal with a reference sample;
- wherein the chemical compound is selected from the group consisting of perfluorinated compounds (PFC), brominated flame retardants (BFRs), polychlorinated biphenyls (PCBs), polychlorinated dibenzo para dioxins (dioxins), polychlorinated dibenzofurans (furans), polycyclic aromatic hydrocarbons (PAHs), toxins from biological origin, metals, food additives, feed additives and combinations thereof, and wherein the chemical compound is not a drug.
The method according to any one of the preceding claims, wherein in step c) the detectable signal is detected and measured by optical techniques, electrochemical techniques, magnetic techniques, mass-based techniques, thermistor-based techniques, and combinations thereof.
The method according to any one of the preceding claims, wherein step d) further comprises determination of the content of the compound.
The method according to any one of the preceding claims, wherein the substrate is selected from the group consisting of fluorescein derivative, coumarin derivative, progesterone, testosterone, caffeine, nicotine, estradiol and clozapine and combinations thereof.
The method according to any one of the preceding claims, wherein the CYP450 enzyme is selected from the group consisting of CYP1, CYP2, CYP3, CYP4, CYP17, CYP21, CYP46, CYP51 and combinations thereof.
The method according to any one of the preceding claims, wherein the chemical compound is a PFC selected from the group consisting of carboxylic acids, sulfonic acids, fluorotelomer alcohols, and combinations thereof.
The method according to claim 35, wherein the substrate is a fluorescein derivative and/or coumarin derivative.
The method according to any one of the preceding claims, wherein the sample is selected from the group consisting of water, a biological sample, a food or feed sample, and an environmental sample.
Kit for carrying out the method according to any of the preceding claims comprising:
- a) a CYP450 enzyme;
- b) a substrate of said CYP450 enzyme, wherein upon a catalytic reaction between the CYP450 enzyme and the substrate a detectable signal is generated;
- c) a buffer solution in a pH of 6-9;
- d) means to detect and measure the detectable signal generated by the catalytic reaction of the CYP450 enzyme and the substrate;
- and
- e) optionally, a NADPH generating system.
Use of a CYP450 enzyme for the detection of a CYP450 enzyme inhibitor chemical compound in a sample, wherein the CYP450 inhibitor chemical compound is not a drug and it is selected from the group consisting of PFC, BFR, PCBs, dioxins, furans, PAHs, toxins from biological origin, metals, food additives, feed additives and combinations thereof, wherein the CYP450 enzyme is selected from of CYP1, CYP2, CYP3, CYP4, CYP17, CYP21, CYP46, CYP51 and combinations thereof, and wherein the sample is water, a biological sample, a food, sample, a feed sample, an environmental sample, or an extract thereof.
Dependencies are inferred from the claim text.