Method for determination of glucose in whole blood and cuvette and photometer for carrying out said method

• US 5,866,349 A
• Filed: 10/16/1991
• Issued: 02/02/1999
• Est. Priority Date: 04/25/1989
• Status: Expired due to Term

• Current Assignee: Jan Evert Lilja, Sven-Erik Lennart Nilsson
• Original Assignee: Jan Evert Lilja, Sven-Erik Lennart Nilsson
• Inventors: Lilja, Jan Evert, Nilsson, Sven-Erik Lennart
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US07/768,255
• Time in Patent Office: 2,666 Days
• Field of Search: 436/14, 435/14, 435/26, 435/13, 422/102
• US Class Current: 435/13
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 2015/011 with lysing, e.g. of erythrocytes
• CPC Class Codes: G01N 2021/0321 One time use cells, e.g. integrally moulded
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/3181 using LEDs
• CPC Class Codes: G01N 21/03 Cuvette constructions
• CPC Class Codes: G01N 21/314 with comparison of measurements at specific and non-specific wavelengths dual wavelength spectrometry G01J3/427
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 2201/1222 Endpoint determination; reaction time determination
• CPC Class Codes: G01N 2201/126 Microprocessor processing
• CPC Class Codes: G01N 33/493 urine

Abstract

A sample of whole blood is contacted with a reagent which by chemical reaction with glucose in the sample brings about a detectable dye concentration change (10) the size of which is determined as a measure of the glucose content of the sample. The sample is initially introduced undiluted in a microcuvette having at least one cavity for receiving the sample. The cavity is internally pretreated with the reagent in dry form, and the chemical reaction takes place in the cavity. Active components of the reagent comprise at least a hemolyzing agent for exposing glucose contained in the blood cells of the sample for allowing total glucose determination, and agents taking part in the chemical reaction and ensuring that the dye concentration change (10) takes place at least in a wavelength range (14) outside the absorption range (12) of the blood hemoglobin. An absorption measurement is performed in said wavelength range directly on the sample in the cuvette. A pretreated disposable cuvette with such a reagent and a photometer are also described.

Claims

1. A method for determining the glucose content in whole blood, in which a sample of undiluted whole blood is contacted with a dye-containing reagent system which undergoes chemical reaction with the glucose in the whole blood sample, the method consisting essentially of the steps of:

introducing the undiluted whole blood sample in a microcuvette having at least one cavity for receiving the sample, said cavity being internally pretreated with the reagent in a dry form and said chemical reaction then taking place in said cavity,

the reagent being comprised of a hemolyzing agent and agents used in the glucose dehydrogenase method, said agents being comprised of glucose dehydrogenase and a redox indicator dye, the hemolyzing agent exposing the glucose contained in the blood cells of the whole blood sample permitting a quantitative total glucose determination in a whole blood hemolysate, the agents which participate in the chemical reaction ensure that a dye concentration change takes place in a wavelength range above 650 nm and,

performing an absorption measurement at said wavelength range directly on the sample in the cuvette, and further conducting a secondary absorption measurement to compensate for background interference in a wavelength range above 700 nm.

2. The method as claimed in claim 1, wherein the reagent is further comprised of diaphorase.

3. The method as claimed in claim 2 wherein the reagent is further comprised of mutarotase.

4. The method as claimed in claim 1 wherein said chemical reaction is an end-point reaction, said absorption measurement being carried out only when said dye concentration change is substantially terminated.

5. A disposable cuvette for carrying out a determination of the glucose content of undiluted whole blood where a sample of undiluted whole blood is contacted with a dye-containing reagent system undergoes chemical reaction with the glucose in the undiluted whole blood sample, the cuvette being comprised of at least one cavity for receiving the undiluted whole blood sample, said cavity being internally pretreated with a reagent in dry form and said chemical reaction occurring in said cavity after introduction of the sample in undiluted form, the reagent being comprised of a hemolyzing agent and agents used in the glucose dehydrogenase method, said agents being comprised of glucose dehydrogenase and a redox indicator dye, the hemolyzing agent exposing the glucose contained in the blood cells of the whole blood sample permitting a quantitative total glucose determination in a whole blood hemolysate, the agents which participate in the chemical reaction ensure that the dye concentration change takes place in a wavelength range above 650 nm and that the cuvette is at least partly transparent for permitting an absorption measurement directly on the sample in the cavity of the cuvette in said wavelength range.

6. A method for determining the glucose content in undiluted whole blood, in which a sample of undiluted whole blood is contacted with a reagent which by chemical reaction with glucose in the undiluted whole blood sample brings about a dye concentration change which is detectable in the sample, whereby the size of the dye concentration change is used to determine the measure of the glucose content, the method consisting essentially of the steps of:

introducing a sample of undiluted whole blood in a microcuvette having at least one cavity for receiving the sample, said cavity being internally pretreated with a reagent in dry form and said chemical reaction with the glucose in the sample taking place in said cavity,

the reagent being comprised of a hemolyzing agent for exposing glucose contained in the blood cells of the sample for allowing a quantitative total glucose determination in a whole blood hemolysate, glucose dehydrogenase, NAD or analog thereof, diaphorase or analog thereof, and a dye, the dye undergoing a concentration change upon said chemical reaction with the reagent corresponding to the blood glucose content of the sample, determining by transmission photometry the concentration change of the dye at a wavelength that is in an absorption range above 650 nm., the dye being selected so that the dye concentration change takes place at least in a wavelength range above 650 nm. which is outside the absorption range of the blood hemoglobin,

and further conducting a secondary absorption measurement to compensate for background interference in a wavelength range above 700 nm.