Abstract 1. A METHOD FOR CONTINUOUS-FLOW ANALYSIS OF OXIDATIVE METABOLITES OF AROMATIC AMINES AND OF COMPOUNDS WHICH PRODUCE FORMALDEHYDE ON OXIDATIVE DEALKYLATION CONSISTING ESSENTIALLY OF THE STEPS OF: (1) COMBINING A FLUID STREAM COMPRISING A SUBSTRATECOFACTOR A PHOSPHATE BUFFER WITH A RAT LIVER HOMOGENATE COMPRISING OXIDATIVE MICROSOMAL ENZYMES, (2) DIVIDING THE RESULTING MIXTURE INTO A FIRST STREAM FOR DETERMING THE QUANTITY OF PROTEIN AND INTO A SECOND STREAM FOR METABOLIZING THE SUBSTRATE AND FOR DETERMINING THE QUANTITY OF METALBOLITE GENERATED DURING SUCH METABOLISM, (3) DILUTING SAID FIRST STREAM WITH PHOSPHATE BUFFER, (4) AERATING SAID DILUTED FIRST STREAM, (5) REACTING SAID AERATED FIRST STREAM WITH A COLOR FORMING REAGENT FOR PROTEIN, (6) COLORIMETRICALLY DETERMINING THE AMOUNT OF PROTEIN, (7) INTERMITTENTLY ADDING SMALL ALIQUOTS OF 100% OXYGEN TO SAID SECOND STREAM, (8) INCUBATING SAID OXYGENATED SECOND STREAM AT A CONTROLLED TEMPERATURE TO PROMOTE THE FORMATION OF METABOLITE BY ENZYME ACTION ON SAID SUBSTRATE, (9) DIALYZING SAID SECOND STREAM AGAINST PHOSPHATE BUFFER, AND (10) COLORIMETRICALLY DETERMINING THE CONCENTRATION OF METABOLITE AFTER REACTING THE DIALSATE WITH A COLOR FORMING REAGENT FOR SAID METABOLITE.
A METHOD FOR CONTINUOUS-FLOW ANALYSIS OF OXIDATIVE METABOLITES OF AROMATIC AMINES AND OF COMPOUNDS WHICH PRODUCE FORMALDEHYDE ON OXIDATIVE DEALKYLATION CONSISTING ESSENTIALLY OF THE STEPS OF: (1) COMBINING A FLUID STREAM COMPRISING A SUBSTRATECOFACTOR A PHOSPHATE BUFFER WITH A RAT LIVER HOMOGENATE COMPRISING OXIDATIVE MICROSOMAL ENZYMES, (2) DIVIDING THE RESULTING MIXTURE INTO A FIRST STREAM FOR DETERMING THE QUANTITY OF PROTEIN AND INTO A SECOND STREAM FOR METABOLIZING THE SUBSTRATE AND FOR DETERMINING THE QUANTITY OF METALBOLITE GENERATED DURING SUCH METABOLISM, (3) DILUTING SAID FIRST STREAM WITH PHOSPHATE BUFFER, (4) AERATING SAID DILUTED FIRST STREAM, (5) REACTING SAID AERATED FIRST STREAM WITH A COLOR FORMING REAGENT FOR PROTEIN, (6) COLORIMETRICALLY DETERMINING THE AMOUNT OF PROTEIN, (7) INTERMITTENTLY ADDING SMALL ALIQUOTS OF 100% OXYGEN TO SAID SECOND STREAM, (8) INCUBATING SAID OXYGENATED SECOND STREAM AT A CONTROLLED TEMPERATURE TO PROMOTE THE FORMATION OF METABOLITE BY ENZYME ACTION ON SAID SUBSTRATE, (9) DIALYZING SAID SECOND STREAM AGAINST PHOSPHATE BUFFER, AND (10) COLORIMETRICALLY DETERMINING THE CONCENTRATION OF METABOLITE AFTER REACTING THE DIALSATE WITH A COLOR FORMING REAGENT FOR SAID METABOLITE.
Claim 2
Independent Claim
177 words
A METHOD FOR CONTINUOUS-FLOW ANALYSIS OF OXIDATIVE METABOLITES OF AROMATIC AMINES AND OF COMPOUNDS WHICH PRODUCE FORMALDEHYDE ON OXIDATIVE DEALKYLATION CONSISTING ESSENTIALLY OF THE STEPS OF: (1) COMBINING A FLUID STREAM COMPRISING A SUBSTRATECOFACTOR A PHOSPHATE BUFFER WITH A RAT LIVER HOMOGENATE COMPRISING OXIDATIVE MICROSOMAL ENZYMES, (2) DIVIDING THE RESULTING MIXTURE INTO A FIRST STREAM FOR DETERMING THE QUANTITY OF PROTEIN AND INTO A SECOND STREAM FOR METABOLIZING THE SUBSTRATE AND FOR DETERMINING THE QUANTITY OF METALBOLITE GENERATED DURING SUCH METABOLISM, (3) DILUTING SAID FIRST STREAM WITH PHOSPHATE BUFFER, (4) AERATING SAID DILUTED FIRST STREAM, (5) REACTING SAID AERATED FIRST STREAM WITH A COLOR FORMING REAGENT FOR PROTEIN, (6) COLORIMETRICALLY DETERMINING THE AMOUNT OF PROTEIN, (7) INTERMITTENTLY ADDING SMALL ALIQUOTS OF 100% OXYGEN TO SAID SECOND STREAM, (8) INCUBATING SAID OXYGENATED SECOND STREAM AT A CONTROLLED TEMPERATURE TO PROMOTE THE FORMATION OF METABOLITE BY ENZYME ACTION ON SAID SUBSTRATE, (9) DIALYZING SAID SECOND STREAM AGAINST PHOSPHATE BUFFER, AND (10) COLORIMETRICALLY DETERMINING THE CONCENTRATION OF METABOLITE AFTER REACTING THE DIALSATE WITH A COLOR FORMING REAGENT FOR SAID METABOLITE.