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US 4,784,945 AGranted
Method for determining lipids
AssigneeBoard of Governors of Wayne State University
InventorsJoseph D. Artiss, Dennis Bozimowski, Robert J. McEnroe, Bennie Zak
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Abstract
A method for determining lipids using thin layer chromatography and multiple enzymatic staining is described. The method uses a hydrolase enzyme, an oxidase enzyme and a peroxidase enzyme in the presence of a chromogen precursor to form an oxidized chromogen stain. The method is particularly useful in determining phospholipids from animal and human tissue and/or fluids.
Claims15
A method for the determination of mixed lipids in a liquid which comprises:
- (a) providing a thin layer chromatographic plate having a stationary phase as the thin layer which separates the mixed lipids;
- (b) applying the liquid containing the mixed lipids to the stationary phase on the plate so that the mixed lipids separate into separated bands of individual lipids upon migration with a mobile phase;
- (c) reacting the individual lipids with an aqueous solution comprising a hydrolase enzyme, an oxidase enzyme, a peroxidase enzyme and chromogen precursor(s), wherein the hydrolase enzyme catalyzes ester hydrolysis products from the lipids, the oxidase enzyme produces hydrogen peroxide by reaction with the hydrolysis products which in turn reacts in the presence of the peroxidase enzyme with the chromogen percursor(s) to form an oxidized chromogen complex(es) which is water insoluble so that the chromogen complex(es) remains in the band on the stationary phase;
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- (d) determining the individual lipids in the thin stationary phase by means of the chromogen complex(es).
2 more elements
Dependencies are inferred from the claim text.