Method for the determination of amylase

Claim 2

A PROCESS FOR DETERMINING THE PANCREATIC A-AMYLASE CONTENT OF A SAMPLE COMPRISING THE STEPS OF ADDING A SUBSTRATE SELECTED FROM THE GROUP CONSISTING OF MALTOTETRAOSE AND MALTOPENTAOSE TO A SOLUTION CONTAINING A MEASURED AMOUNT OF THE SAMPLE AND DETERMINED THE GLUCOSE EVOLVED BY THE REACTION BETWEEN THE PANCREATIC A-AMYLASE AND THE SUBSTRATE WHILE MAINTAINING THE SOLUTION AT A SUBSTANTIALLY CONSTANT TEMPERATURE AND A SUBSTANTIALLY CONSTANT PH.

Claim 3

A process for determining the pancreatic Alpha -amylase content of a sample comprising the steps of adding a substrate selected from the group consisting of maltotetraose and maltopentaose to a solution containing a measured amount of the sample and determining the glucose evolved by the reaction between the pancreatic Alpha -amylase and the substrate while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 4

A process for determining the pancreatic Alpha -amylase content of a sample comprising the steps of adding a substrate selected from the group consisting of maltotetraose and maltopentaose to a solution containing a measured amount of the sample and determining the glucose evolved by the reaction between the pancreatic Alpha -amylase and the substrate while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 5

The process of claim 1 wherein Beta -glycerol phosphate is added to the solution as a buffer.

Claim 6

The process of claim 2 wherein the substrate is maltotetraose.

Claim 7

The process of claim 3 wherein the step of determining the glucose evolved is accomplished by a coupled enzyme reaction.

Claim 8

The process of claim 3 wherein the step of determining the glucose evolved is accomplished by measuring the rate at which glucose is evolved.

Claim 9

A process for determining the total Alpha -amylase content of a sample comprising the steps of adding a substrate selected from the group consisting of maltotetraose and maltopentaose to a solution containing a measured amount of said sample and non-rate limiting amounts of a maltase; and determining the glucose evolved by the reactions which take place in the solution, while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 10

A process for determining the total Alpha -amylase content of a sample comprising the steps of adding a substrate selected from the group consisting of maltotetraose and maltopentaose to a solution containing a measured amount of said sample and non-rate limiting amounts of a maltase; and determining the glucose evolved by the reactions which take place in the solution, while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 11

The process of claim 6 wherein Beta -glycerol phosphate is added to the solution as a buffer.

Claim 12

The process of claim 7 wherein the substrate is maltotetraose.

Claim 13

The process of claim 7 wherein the step of determining the glucose evolved is accomplished by a coupled enzyme reaction.

Claim 14

The process of claim 7 wherein the step of determining the glucose evolved is accomplished by measuring the rate at which glucose is evolved.

Claim 15

The process of claim 7 wherein said maltase is Alpha -glucosidase.

Claim 16

The process for determining the salivary Alpha -amylase content of a sample comprising the steps of determining the total Alpha -amylase content of the sample by the process of claim 7; determining the panceratic Alpha -amylase content of the sample by the process of adding a substrate selected from the group consisting of maltotetraose and maltopentaose to a solution containing a measured amount of the sample and determining the glucose evolved by the reaction between the pancreatic Alpha -amylase and the substrate while maintaining the solution at a substantially constant temperature and a substantially constant pH; and subtracting the two values.

Claim 17

A process for determining the Alpha -amylase content of a sample comprising the steps of: adding a measured amount of the sample to a solution containing non-rate limiting amounts of hexokinase, glucose-6-phosphate dehydrogenase, sodium chloride, magnesium sulphate, and Beta -nicotinamide-adenine dinucleotide; adding to the solution Alpha -glucosidase and maltotetraose; and determining the rate at which glucose is evolved while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 18

A process for determining the Alpha -amylase content of a sample comprising the steps of: adding a measured amount of the sample to a solution containing non-rate limiting amounts of hexokinase, glucose-6-phosphate dehydrogenase, sodium chloride, magnesium sulphate, and Beta -nicotinamide-adenine dinucleotide; adding to the solution Alpha -glucosidase and maltotetraose; and determining the rate at which glucose is evolved while maintaining the solution at a substantially constant temperature and a substantially constant pH.

Claim 19

The process of claim 13 wherein Beta -glycerol phosphate is added to the solution as a buffer.