Sphingomyelin measurement method and measurement kit
AssigneeKyowa Medex Co., Ltd.
InventorsTakehide Kimura, Kazuhito Miyauchi, Hideyuki Kuwata
Life of this patent
Estimating the term…
Abstract
Provided is a method for simply and accurately measuring sphingomyelin in a sample, and a kit therefor. The method is a method for measuring sphingomyelin in a sample, comprising reacting the sample with a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine, a lysophospholipase or a monoglyceride lipase, and a choline oxidase, eliminating the formed hydrogen peroxide, reacting the resultant with a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin, and a choline oxidase, and measuring the formed hydrogen peroxide.
Claims32
A method for measuring sphingomyelin in a sample, comprising
- reacting the sample with a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- and a choline oxidase,
- eliminating the formed hydrogen peroxide,
- reacting the resultant with a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- and a choline oxidase, and
- measuring the formed hydrogen peroxide.
A method for measuring sphingomyelin in a sample, comprising
- reacting the sample with a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- an oxidized coenzyme;
- a choline dehydrogenase;
- and a reduced coenzyme oxidase,
- eliminating the formed hydrogen peroxide,
- reacting the resultant with a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- an oxidized coenzyme;
- a choline dehydrogenase;
- and a reduced coenzyme oxidase, and
- measuring the formed hydrogen peroxide.
A kit for measuring sphingomyelin in a sample, comprising
- a first reagent comprising a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- a choline oxidase;
- and a catalase, and
- a second reagent comprising a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- and a catalase inhibitor.
A kit for measuring sphingomyelin in a sample, comprising
- a first reagent comprising a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- a choline oxidase;
- a peroxidase;
- and one of a pair oxidative coupling-coloring chromogens, and
- a second reagent comprising a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- and the other of the pair of oxidative coupling-coloring chromogens.
A kit for measuring sphingomyelin in a sample, comprising
- a first reagent comprising a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- a choline dehydrogenase;
- an oxidized coenzyme;
- a reduced coenzyme oxidase;
- and a catalase, and
- a second reagent comprising a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- and a catalase inhibitor.
A kit for measuring sphingomyelin in a sample, comprising
- a first reagent comprising a phospholipase D which does not react with sphingomyelin and lysophosphatidylcholine but reacts with phosphatidylcholine;
- a lysophospholipase or a monoglyceride lipase;
- a choline dehydrogenase;
- an oxidized coenzyme;
- a reduced coenzyme oxidase;
- a peroxidase;
- and one of a pair of oxidative coupling-coloring chromogens, and
- a second reagent comprising a phospholipase D which does not react with glycerol-3-phosphorylcholine and free fatty acid but reacts with sphingomyelin;
- and the other of the pair of oxidative coupling-coloring chromogens.
Dependencies are inferred from the claim text.