Granted Patent B2

US 9,976,172 · App. 15/052,914 · Granted May 22, 2018

Abstract

Claims (16)

1. A method for analyzing reactions, comprising:

(a) providing a solution comprising solvent, at least one acceptor chemical and at least one donor chemical, wherein the donor chemical is capable of donating a chemical moiety comprising a chemical element to the acceptor chemical, and at least two controller chemicals capable of affecting the reaction between the donor chemical and the acceptor chemical;

(b) incubating the solution to allow a portion of the acceptor chemical to react with the donor chemical to form an acceptor product;

(c) separating at least a portion of the unreacted donor chemical from the acceptor product;

(d) removing at least a portion of the solvent before measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical; and

(e) measuring using x-ray fluorescence at least one of the acceptor product or the donor chemical, wherein:

the controller chemicals are enzyme inhibitors; and

the sample of donor chemical or acceptor product that is subjected to x-ray fluorescence measurement is concentrated such that the area containing more than 50% of the sample based on the total amount of the sample of donor chemical or acceptor product that is subjected to x-ray fluorescence measurement is less than 0.005 square centimeters.

2. The method of claim 1 , wherein the step of removing at least a portion of the solvent is performed using reduced pressure.

3. The method of claim 1 , wherein at least 80% of the solvent is removed.

4. The method of claim 3 , wherein the step of separating at least a portion of the unreacted donor chemical from the acceptor product is performed using chromatography.

5. The method of claim 3 , wherein the step of separating at least a portion of the unreacted donor chemical from the acceptor product is performed using a centrifuge.

6. The method of claim 3 , wherein the step of separating at least a portion of the unreacted donor chemical from the acceptor product is performed using size exclusion chromatography.

7. The method of claim 1 , wherein the reaction between the donor chemical and the acceptor chemical forms at least one chemical bond.

8. The method of claim 1 , wherein the reaction between the donor chemical and the acceptor chemical breaks at least one bond.

9. The method of claim 1 , wherein the enzyme inhibitor is selected from the group consisting of phthalene-1-sulfonyl-1H-hexahydro-1,4-diazepine; 1-(5-isoquinolinesulfonyl)-1H-hexahydro-1,4-diazepine; 1-(5-isoquinolinesulfonyl)-2-methylpiperazine; 1-(5isoquinolinesulfonyl)-piperazine; 1,2-Dimethoxy-N-methyl(1,3) benzodioxolo(5,6-c) phenanthridinium chloride; 1-[N,0-bis-(5isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenyl piperazine; 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; 2,3-Dihydro-N,N-dimethyl-2-oxo-3-[(4,5,6,7-tetrahydro-1H-indol -2-yl)methylene]-1H-indole-5-sulfonamide; 2,5-Dihydroxymethylcinnamate; 2-Cyano-N-(2,5-dibromophenyl)-3-hydroxy-2-butenamide; 3-[[4(dimethylamino)phenyl]methylene]-1,3-dihydro-2H-indol-2-one; 4-(2,5-Dihydroxy-benzylamino)benzoic Acid Adamantan-1-yl Ester; 4-(3-chloro-4-fluoroanilino)-7-methoxy-6-(3morpholinopropoxy)quinazoline; 4,5,6,7-Tetrabromo-2-azabenzimidazole;4-[(3-Bromophenyl)amino]-6,7-dimethoxyquinazoline; 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4,d]pyrimidine; 4-Amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine; 4-Amino-5-iodo-7-(beta-D-ribofuranosyl)pyrrolo[2,3d]-pyrimidine; 4′-Amino-6-Hydroxyflavone; 4-Amino-N-(2,5-dihydroxybenzyl)adamantyl Benzoate; 4-Amino-N-(2,5-dihydroxybenzyl)methylbenzoate; 5,6-Dichloro-1-b-D-ribofuranosylbenzimidazole; 5,7-Dimethoxy-3-(4-pyridinyl)quinoline; 6-Amino-4-methyl-8-([8-D-ribofuranosyl)4H,8H-pyrrolo[4,3,2-de]pyrimido[4,5-c]pyridazine; Bisindolylmaleimide IX; N-(2′-Guanidinoethyl)-5-isoquinolinesulfonamide; N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide; N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide; N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide; N-Benzyl-3,4-dihydroxy-enzylidenecyanoacetamide; Picropodophyllin; Quercetin; rac-2-methyl-1-octadecyl-glycero-(3)-phosphocholine; Rapamycin; Rottlerin; Staurosporine; trans-3,3′,4,5′-Tetrahydroxystilbene; 4-[(1E)-2-(3,5-Dihydroxyphenyl)ethenyl]-1,2-benzenediol; trans-4-[(1R)-1-Aminoethyl]N4pyridinylcyclohexanecarboxamide and wortmannin.

Assignments (15)