Method of detecting sugars

• US 6,586,195 B1
• Filed: 11/19/2001
• Issued: 07/01/2003
• Est. Priority Date: 11/19/2001
• Status: Expired due to Fees

• Current Assignee: R.E. Davis Chemical Corporation
• Original Assignee: R.E. Davis Chemical Corporation
• Inventors: Friedman, Arthur J.
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US10/015,216
• Time in Patent Office: 589 Days
• Field of Search: 435/18, 435/14, 435/4, 435/967, 435/3
• US Class Current: 435/18
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 2333/924 acting on glycosyl compounds (3.2)
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems

Abstract

A method for detecting reducing sugars in samples using a test reagent containing an indicator capable of producing calorimetric changes when contacted with reducing sugars. Under alkaline conditions, reducing sugars present in a sample affect a reduction of the indicator to a colorless or differently colored reduced form. The method may include the additional step of pretreating the sample with a glycosidase to convert non-reducing sugars to reducing sugars.

Claims

1. A method of detecting reducing sugars in a sample comprising the steps of:

contacting a test reagent with the sample, the test reagent having an alkaline pH and including an indicator capable of providing a colorimetric change when contacted with reducing sugar in the sample;

subsequently detecting a colorimetric change in the indicator indicating the presence of reducing sugars in the sample.

2. A method according to claim 1 wherein the indicator is a phenazine compound.

3. A method according to claim 2 wherein the phenazine compound selected from the group consisting of azocarmine B, indoine blue, methylene violet 3RAX, safranine O, phenosafranin, and Janus Green B.

4. A method according to claim 1 wherein the test reagent further comprises a base selected from the group consisting of NaOH, KOH, Sr(OH)₂, Ba(OH)₂, and NH₄OH.

5. A method according to claim 1 including the additional step of pretreating the sample with a glycosidase prior to contacting the sample with the test reagent.

6. A method according to claim 5 wherein the glycosidase is invertase.

7. A method according to claim 1 wherein the indicator is pre-adsorbed onto an adsorbent material to form a pre-adsorbed material and the pre-adsorbed material is added to an alkaline solution to generate the test reagent prior to contacting the test reagent with the sample.

8. A method of detecting reducing sugars in a sample comprising the steps of

contacting a test reagent with the sample, the test reagent having an alkaline pH and including a phenazine compound capable of providing a calorimetric change when contacted with reducing sugar in the sample;

subsequently detecting a calorimetric change in the phenazine compound indicating the presence of reducing sugars in the sample.

9. A method of detecting reducing sugars in a sample comprising the steps of:

contacting a test reagent with the sample, the test reagent having an alkaline pH and including an indicator selected from the group consisting of azocarmine B, indoine blue, methylene violet 3 RAX, safranine O, phenosafranine, and Janus Green B, the indicator capable of providing a calorimetric change when contacted with reducing sugar in the sample;

subsequently detecting a colorimetric change in the indicator indicating the presence of reducing sugars in the sample.

10. A method of detecting sugars comprising the steps of:

pretreating the sample with a glycosidase to form a pretreated sample;

contacting a test reagent with the pretreated sample, the test reagent having an alkaline pH and including a phenazine compound capable of providing a calorimetric change when contracted with reducing sugar within the sample;

subsequently detecting a colorimetric change in the indicator indicating the presence of reducing sugars in the sample.