Method of determining lipase activity using a transparent stable aqueous solution of triglyceride substrate
AssigneeIatron Laboratories, Inc.
InventorsIyoko Miki, Hiroyuki Tsubota, Toshio Tsuchiko, Hisashi Ochi
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Abstract
A transparent and stable aqueous solution of a substrate for a determination of lipase, comprising triglyceride uniformly solubilized therein, a method for determination of lipase activity in a sample, employing the aqueous solution, and a process for the manufacture of a lyophilized product capable of forming the aqueous solution are disclosed. A process for the manufacture of a transparent and stable aqueous solution of triglyceride is also disclosed.
Claims2
A method of determining human pancreatic lipase in a sample, comprising the steps of:
- (1) placing the sample in contact with a transparent aqueous solution containing a triglyceride as a substrate, in the presence of a compound which accerates lipase activity and hydrolyzes the triglyceride, thus liberating fatty acids;
- and
- (2) measuring the amount of the liberated fatty acids,
- wherein the transparent aqueous solution containing the triglyceride is prepared by the steps of
- (a) adding the triglyceride to an aqueous solution containing the nonionic surfactant to form a mixture;
- (b) heating the mixture to or above the cloud point temperature of the nonionic surfactant containing aqueous solution while stirring the mixture;
- (c) cooling the mixture to a temperature below the cloud point while stirring to form a transparent solution;
- (d) lyophilizing the transparent solution in the presence of an excipient or stabilizer selected from the group consisting of sucrose, raffinose, lactose, dextran, carboxymethyl cellulose, FICOLL.RTM., albumin, glycine and comginations thereof;
- and
- (e) dissolving the lyophilizate in an aqueous solution.
A method of determining human pancreatic lipase in a sample, comprising the steps of:
- (1) placing the sample in contact with a transparent aqueous solution containing a triglyceride as substrate in the presence of a compound which accerates lipase activity and hydrolyzes the triglyceride, thus liberating fatty acids;
- and
- (2) measuring the amount of the liberated fatty acids,
- wherein the transparent aqueous solution containing the triglyceride has been prepared by the steps of:
- (a) adding the triglyceride to an aqueous solution containing a nonionic surfactant to form a mixture;
- (b) heating the mixture to or above the cloud point temperature of the nonionic surfactant containing aqueous solution while stirring the mixture;
- (c) cooling the mixture to a temperature below the cloud point while stirring to form a transparent solution;
- (d) lyophilizing the transparent solution in the presence of an excipient or stabilizer selected from the consisting of sucrose, raffinose, lactose, dextran, carboxymethyl cellulose, FICOLL.RTM., albumin, glycine and combinations thereof;
- and
- (e) dissolving the lyophilizate in an aqueous solution.
Dependencies are inferred from the claim text.