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Enzymatic colorimetric method and reagent for measuring concentration of glycocholic acid

A technology of glycocholic acid and detection method, which is applied to the measurement of color/spectral characteristics, etc., can solve the problems of poor anti-interference performance and low accuracy, and achieve the effects of low cost, high accuracy, and simple and easy-to-use methodology.

Inactive Publication Date: 2018-05-25
王学忠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention fundamentally solves the limitations of the immunoturbidimetric method and the homogeneous enzyme immunoassay
By establishing a new enzymatic colorimetric method, which does not have the difficulty of preparing antibody materials in immunological methods, it solves the defects of low accuracy and poor anti-interference performance in the existing clinical automatic reagent detection applications ; And provide a kind of test kit based on the mensuration glycocholic acid concentration of this method

Method used

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  • Enzymatic colorimetric method and reagent for measuring concentration of glycocholic acid
  • Enzymatic colorimetric method and reagent for measuring concentration of glycocholic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Preparation of Reagent 1:

[0044] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 1:

[0045] 100mM / L Tris buffer pH 7.90

[0046] 50mM / L sodium chloride

[0047] 1mM / L EDTA

[0048] 20mM / L Mannitol

[0049] 0.02% Proclin-300

[0050] 0.1% Tween-20

[0051] 0.2% bovine serum albumin

[0052] 4MU / L catalase

[0053] 5KU / L ascorbate oxidase

[0054] 20KU / L glycine oxidase

[0055] 0.3mM / L 4-AA

[0056] Preparation of Reagent 2:

[0057] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 2:

[0058] 100mM / L Tris buffer pH7.50

[0059] 0.15% sodium azide

[0060] 100mmol / L Mannitol

[0061] 50mM / L sodium chloride

[0062] 10mM magnesium sulfate

[0063] 1mM / L EDTA

[0064] 0.1% Tween-20

[0065] 0.2% bovine serum albumin

[0066] 2KU / L peroxidase

[0067] 50KU / L glycocholic acid hydrolase

[0068] 10mM / L TBHBA

Embodiment 2

[0070] Preparation of Reagent 1:

[0071] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 1:

[0072] 100mM / L Tris buffer pH 7.90

[0073] 100mM / L sodium chloride

[0074] 5mM / L EDTA

[0075] 100mM / L Mannitol

[0076] 0.02% Proclin-300

[0077] 0.3% Tween-20

[0078] 0.5% bovine serum albumin

[0079] 5MU / L catalase

[0080] 5KU / L ascorbate oxidase

[0081] 50KU / L glycine oxidase

[0082] 0.2mM / L SMBTH

[0083] Preparation of Reagent 2:

[0084] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 2:

[0085] 100mM / L Tris buffer pH7.50

[0086] 0.15% sodium azide

[0087] 100mmol / L Mannitol

[0088] 50mM / L sodium chloride

[0089] 15mM magnesium sulfate

[0090] 2mM / L EDTA

[0091] 0.3% Tween-20

[0092] 0.2% bovine serum albumin

[0093] 2KU / L peroxidase

[0094] 100KU / L Glycocholic Acid Hydrolase

[0095] 2mM / L ALPS

Embodiment 3

[0097] Preparation of Reagent 1:

[0098] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 1:

[0099] 50mM / L 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid buffer pH 8.00

[0100] 50mM / L sodium chloride

[0101] 2mM / L EDTA

[0102] 100mM / L Mannitol

[0103] 0.02% Proclin-300

[0104] 0.1% Triton X-100

[0105] 0.2% bovine serum albumin

[0106] 5MU / L catalase

[0107] 5KU / L ascorbate oxidase

[0108] 5KU / L glycine oxidase

[0109] 0.3mM / L CCAP

[0110] Preparation of Reagent 2:

[0111] Dissolve the following substances in pure water in sequence according to each concentration to prepare reagent 2:

[0112] 100mM / L 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid buffer pH8.00

[0113] 0.15% sodium azide

[0114] 100mmol / L Mannitol

[0115] 50mM / L sodium chloride

[0116] 10mM magnesium sulfate

[0117] 1mM / L EDTA

[0118] 0.1% Triton X-100

[0119] 0.2% bovine serum albumin

[0120] 2KU / L pe...

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Abstract

The invention discloses an enzymatic detection method for measuring content of glycocholic acid in a sample by adopting glycocholic acid hydrolase and glycine oxidase. With the adoption of the method,a brand-new enzymatic colorimetric reagent is further provided and applied to measurement of glycocholic acid in clinical body fluid, and the reagent has the characteristics of being high in specificity and accuracy and convenient to use and adopting easily available materials due to adoption of specific enzyme.

Description

technical field [0001] The invention belongs to the technical field of substance inspection and determination, and in particular relates to an enzymatic colorimetric method for measuring the concentration of glycocholic acid and a reagent thereof. Background technique [0002] Cholylic acid (Cholyglycine, CG) is one of the conjugated cholic acids formed by the combination of bile acid and glycine. The normal metabolic pathway of CG is the entero-hepatic circulation, which is synthesized by liver cells, discharged into the gallbladder through the capillary and bile ducts, and enters the duodenum with bile to help digest food. 95% of bile acid is reabsorbed in the terminal ileum, and then returned to the liver through the portal vein, where it is taken up and reused by liver cells. In the serum, it mainly exists in the form of protein binding, and the total amount spilled into the systemic circulation is less than 1%. Under normal circumstances, the bile acid content in peri...

Claims

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Application Information

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IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 不公告发明人
Owner 王学忠
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