High-concentration glucose removal method and kit for determining serum 1,5 anhydro-D-glucitol based on pyranose oxidase method
An anhydroglucitol and glucose technology, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, etc., can solve problems such as the inability of glucose elimination methods to be applied.
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Embodiment 1
[0018] Precisely formulated glucose scavenging reagent (hereinafter referred to as "Clearing Reagent 1") with the following composition: pH6.50 MES buffer is 20mmol / L, GK is 1KU / L, G6PD is 5KU / L, NADP is 5mmol / L, and PK is 0.5KU / L, PEP is 1mmol / L, ATP is 0.2mmol / L. Perform a glucose clearance test to determine the effectiveness of this method. Add (1) deionized water, (2) test solution containing 304μmol / L 1,5-AG, (3) test solution containing 60mmol / L glucose, and (4) test solution containing 304μmol / L glucose into 4 test tubes. L 1,5-AG and 60mmol / L glucose test solution each 0.10ml, and then respectively inject 1.50ml of the above glucose removal reagent, after heating at 37℃ for 5 minutes, each tube containing 4-aminoantipyrine ( 4-AAP) 2mmol / L, PROD50KU / L, peroxidase (HRP) 6KU / L, 3-hydroxy-2,4,6-tribromobenzoic acid (TBHBA) 2.0mmol / L color developer each 0.75 Milliliter, continue heating at 37°C for 5 minutes, and measure the absorbance at 510 nm to obtain the results in ...
Embodiment 2
[0023] Precisely formulated glucose scavenging reagent (hereinafter referred to as "Clearing Reagent 2") with the following composition: pH6.50 MES buffer is 1000mmol / L, GK is 25KU / L, G6PD is 100KU / L, NADP is 30mmol / L, and PK is 10KU / L, PEP is 2mmol / L, ATP is 1mmol / L. The glucose clearance test was performed as in Example 1 to determine the effectiveness of this method. Add (1) deionized water, (2) test solution containing 304μmol / L 1,5-AG, (3) test solution containing 60mmol / L glucose, and (4) test solution containing 304μmol / L glucose into 4 test tubes. 0.10ml each of 1,5-AG and 60mmol / L glucose to be tested, then add 1.50ml of glucose scavenging reagent, after heating at 37℃ for 4 minutes, add 0.75ml each of the color reagent consistent with Example 1 , Continue heating at 37°C for 5 minutes, and measure the absorbance at 510 nm to obtain the results in Table 3.
[0024] table 3
[0025] Test solution Absorbance values (1) Deionized water 0.033 (2) 304μmol / L1,5-AG ...
Embodiment 3
[0028] Precisely formulated glucose scavenging reagent (hereinafter referred to as "clearing reagent 3") with the following composition: pH6.50 MES buffer is 2000mmol / L, GK is 50KU / L, G6PD is 200KU / L, NADP is 60mmol / L, and PK is 20KU / L, PEP is 3mmol / L, ATP is 2mmol / L. The glucose clearance test was performed as in Example 1 to determine the effectiveness of this method. Add (1) deionized water, (2) test solution containing 304μmol / L 1,5-AG, (3) test solution containing 60mmol / L glucose, and (4) test solution containing 304μmol / L glucose into 4 test tubes. 0.10ml each of 1,5-AG and 60mmol / L glucose to be tested, then add 1.50ml of glucose scavenging reagent, after heating at 37℃ for 3 minutes, add 0.75ml each of the color reagent consistent with Example 1 , Continue heating at 37°C for 5 minutes, and measure the absorbance at 510 nm to obtain the results in Table 4.
[0029] Table 4
[0030] Test solution Absorbance values (1) Deionized water 0.037 (2) 304μmol / L1,5-AG...
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