Process to extract gluten from food products and kit for achieving same
View Patent ↗The invention relates to a method for extracting the gluten contained in a sample using an ethanol aqueous solution in the presence of a composition containing a disulphur group reducer and a decoupling agent, in a buffer having a pH of between 7 and 8. The inventive method can be used to extract quantitatively the gluten contained in a heat-processed or non-heat processed food sample before the quantification of the gluten by ELISA. The method is suitable for food analyses, in particular, for foodstuffs intended for celiac sufferers.
1. A process for extracting gluten from a food sample comprising extracting the gluten contents of said sample with an aqueous ethanol solution in the presence of a composition consisting of a disulphide group reducing agent and a dissociating agent in buffer with pH between 7 and 8, wherein the extraction process does not include contact between the food sample and any alcohol other than said ethanol.
2. The process of claim 1 , wherein said aqueous ethanol solution has an ethanol content between 50 and 70%.
3. The process of claims 1 , wherein said extraction comprises:
(a) mixing said sample with the composition consisting of a disulphide group reducing agent and a dissociating agent in a buffer with pH between 7 and 8 to make a mixture;
(b) incubating the mixture at a temperature of between 37 and 50° C., for a period of 30 to 60 minutes;
(c) cooling the mixture to room temperature;
(d) adding said aqueous ethanol solution; and
(e) separating the supernatant that contains the extracted gluten.
4. The process of claim 3 , wherein said sample with the composition is incubated at a temperature of 50° C. for 40 minutes.
5. The process of claim 3 , wherein said aqueous ethanol solution is an aqueous ethanol solution containing 60% of ethanol.
6. The process of claim 3 further comprising incubating the mixture after step (d).
7. The process of claim 6 , wherein the incubation of the mixture after step (d) is done at room temperature for 1 hour.
8. A method to quantify by ELISA the amount of gluten present in a sample of food that comprises extracting said gluten from said sample using the process of claim 1 and quantifying by ELISA the gluten extracted.
9. A kit comprising a composition consisting of a disulphide group reducing agent and a disassociating agent in buffer with pH between 7 and 8 partially mixed or separate and an aqueous ethanol solution or separate components of the aqueous ethanol solution, wherein the kit does not include any alcohol except for said ethanol.
10. The kit of claim 9 , further comprising ELISA reagents necessary to quantify gluten content of a food product, wherein said reagents are completely mixed, partially mixed or separate.
11. The process of claim 1 , wherein the disulphide group reducing agent is selected from the group consisting of 2-mercaptoethanol, dithiotreitol and mixtures thereof.
12. The process of claim 1 , wherein the disassociating agent is selected from the group consisting of guanidine hydrochloride, urea and mixtures thereof.
13. The process of claim 1 wherein the disulphide group reducing agent is 2-mercaptoethanol and the disassociating agent is guanidine hydrochloride.
14. The kit of claim 9 , wherein the disulphide group reducing agent is selected from the group consisting of 2-mercaptoethanol, dithiotreitol and mixtures thereof.
15. The kit of claim 9 , wherein the disassociating agent is selected from the group consisting of guanidine hydrochloride, urea and mixtures thereof.
16. The kit of claim 9 wherein the disulphide group reducing agent is 2-mercaptoethanol and the disassociating agent is guanidine hydrochloride.
17. The kit of claim 10 , wherein the disulphide group reducing agent is selected from the group consisting of 2-mercaptoethanol, dithiotreitol and mixtures thereof.
18. The kit of claim 10 , wherein the disassociating agent is selected from the group consisting of guanidine hydrochloride, urea and mixtures thereof.
19. The kit of claim 10 wherein the disulphide group reducing agent is 2-mercaptoethanol and the disassociating agent is guanidine hydrochloride.