Determination reagent for glycocholic acid and preparation method of determination reagent

A technique for glycocholic acid and a reagent, which is applied in the field of glycocholic acid determination reagent and its preparation, can solve the problems of poor quantitative accuracy, long operation time, low sensitivity and the like, and achieves good accuracy, high specificity and good stability. Effect

Inactive Publication Date: 2016-05-18
浙江达美生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] (1) Latex-enhanced immunoturbidimetric method, which is less sensitive and prone to prozone phenomenon
[0013] (2) Chemiluminescence method, although the method has high sensitivity, it is unstable and has poor repeatability
[0014] (3) Enzyme-linked immunoassay, which has poor quantitative accuracy, long operation time and low degree of automation, and is generally only used for qualitative detection

Method used

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  • Determination reagent for glycocholic acid and preparation method of determination reagent
  • Determination reagent for glycocholic acid and preparation method of determination reagent
  • Determination reagent for glycocholic acid and preparation method of determination reagent

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Embodiment Construction

[0050] The main reagents used in this embodiment: phosphate buffer: pH7.0, 0.65mmol / L; Thio-NAD (thio-oxidized coenzyme I): 1g / L; glycocholic acid dehydrogenase (GCDH): ≥5000U / L L: sodium azide: 0.6mmol / L; NADH (reduced coenzyme I): 6g / L; all purchased from Enzymaker.

[0051] 1) Preparation of R1 reagent

[0052] ①Pour 1500ml of purified water into a suitable container.

[0053] ② Turn on the electric stirrer, and the stirring speed is 450 rpm.

[0054] ③Weigh 1g of Tris and add it into the above purified water, stir until completely dissolved.

[0055] ④Weigh 1g of thiooxidized coenzyme I and add it to the above solution, stir until completely dissolved.

[0056] ⑤ Stir for more than ten minutes.

[0057] ⑥Set the volume to 1600ml.

[0058] 2) Preparation of R2 reagent

[0059] ①Pour 300ml of purified water into a suitable container.

[0060] ② Turn on the electric stirrer, and the stirring speed is 450 rpm.

[0061] ③ Weigh 0.3g Tris and add it into the above purified...

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Abstract

The invention relates to a determination reagent for glycocholic acid and a preparation method of the determination reagent, and belongs to the technical field of biochemical means testing. The determination reagent comprises an R1 reagent, an R2 reagent and a glycocholic acid standard sample; a solution formed by dissolving a sulfo-oxidation type coenzyme into purified water added with a buffer solution is adopted as the R1 reagent; a solution formed by dissolving a reduction type coenzyme into purified water added with a buffer solution, glycocholic acid dehydrogenase and sodium azide is adopted as the R2 reagent; a solution formed by dissolving a sodium glycocholate hydrate, a buffer solution and sodium azide into purified water is adopted as the glycocholic acid standard sample. The determination reagent is applied to glycocholic acid determination and has the advantages of being quick, sensitive, good in accuracy, high in specificity, good in stability and the like.

Description

technical field [0001] The invention relates to a reagent for measuring glycocholic acid and a preparation method thereof, belonging to the technical field of biochemical means testing. Background technique [0002] Serum Cholyglycine (CG) is one of the conjugated bile acids formed by the combination of bile acid and glycine. In liver cells, cholesterol is transformed into primary bile acid through extremely complex enzymatic reactions. Among them are cholic acid (CA) and chenodeoxycholic acid (CD-CA). There are three hydroxyl groups (C3, C7, C12) on the steroid core of cholic acid, and the hydroxyl group at the end of the side chain is combined with glycine through a peptide bond, with a molecular weight of 462u. [0003] CG is synthesized in the liver cells, discharged into the gallbladder through the bile duct for storage, the gallbladder contracts after meals, and is discharged into the small intestine along with the bile to participate in the digestion and absorption o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 方朝君
Owner 浙江达美生物技术有限公司
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