Antioxidant for coagulative reagent

A technology of antioxidants and reagents, applied in biological testing, material inspection products, etc., can solve problems such as unfavorable safety production, phenol impact, etc.

Active Publication Date: 2015-10-21
QINGDAO GUGAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the destructive effects of phenol on human health and the environment, and it is not conducive to safe production, its application is limited

Method used

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  • Antioxidant for coagulative reagent
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  • Antioxidant for coagulative reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Effect of Salvianolic Acid A on the Antioxidant Activity of APTT Reagent

[0038] (1) Reagent oxidation resistance test at room temperature

[0039] Preparation of APTT reagent: Take mixed phospholipids (PS of 20 μmol, PC of 35 μmol and PE of 35 μmol), join in 50mL concentration and be the HEPES buffer solution of 25mmol / L, the sodium azide of 0.05% is contained in the HEPES buffer solution, 100mM Sodium chloride and 20mg / L of butylated hydroxytoluene, the pH is 7.4. Add bovine serum albumin, 40nm silicon dioxide, mannitol and maltose to the mixed solution, make up the filtrate to a volume of 1L with deionized water, so that the composition and content of each part in the final solution are: 0.07mmol mixed Phospholipids, 3g silicon dioxide, 0.7g butylated hydroxytoluene, 25g mannitol, 20g maltose, 5g bovine serum albumin, 30g sodium chloride and 0.1g sodium azide. After the above mixed solution is uniformly mixed by a magnetic stirrer, it is filtered with a 0.22 micro...

Embodiment 2

[0053] Effects of Salvianolic Acid B on the Antioxidation of PT Reagents

[0054] (1) Reagent oxidation resistance test at room temperature

[0055] Preparation of PT reagent: Take mixed phospholipids (PS of 20 μmol, PC of 35 μmol and PE of 35 μmol), join in 50mL concentration and be 25mmol / L in the HEPES buffer solution, the sodium azide of 0.05% is contained in the HEPES buffer solution, 100mM Sodium chloride and 20mg / L of butylated hydroxytoluene, the pH is 7.4. Pour the buffer solution into 1L burnout, add 10g TritonX-100, and stir gently at 25°C for 90 minutes to obtain a uniformly dispersed phospholipid solution. Add rabbit brain tissue factor, bovine serum albumin, methionine, sodium chloride, PEG, mannitol, maltose, and sodium azide to the above solution containing phospholipids, and set the volume to 1 L, so that the volume of each component in the final solution The content is: rabbit brain tissue factor 300nm, mixed phospholipids 2.5mM, bovine serum albumin 0.1%, ...

Embodiment 3

[0069] Effect of Rosmarinic Acid on the Antioxidant Activity of TT Reagent

[0070] (1) Reagent oxidation resistance test at room temperature

[0071] Prepare a buffer solution with a Tris concentration of 50 mM (pH7.4), and add a final concentration of 2% BSA, 0.05% sodium azide, 3.5% methionine, 1.5% PEG, 3% Mannitol, 2.5% maltose, and 10 mM calcium chloride, all of which except calcium chloride are mass volume fractions. The above mixed solution was filtered with a 0.22 micron filter membrane, and a freeze-dried preparation of human thrombin was added to the filtrate so that the concentration of human thrombin in the final filtrate was 25 U / ml to obtain TT reagent. The TT value of the reagent was determined by CA1500 coagulation analyzer to be 15.8s.

[0072] Contrast reagents: Studies have shown that in the blood coagulation preparations containing thrombin, the addition of high concentrations of salvianolic acid and rosmarinic acid not only did not play an antioxidant r...

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Abstract

The invention relates to an antioxidant for a coagulative reagent. The antioxidant is salvianolic acid or rosmarinic acid, and the salvianolic acid comprises salvianolic acid A, salvianolic acid B and salvianolic acid C. The coagulative reagent comprises an APTT reagent, a PT reagent, a TT reagent, an FIB reagent and an AT-III reagent. The concentration of the salvianolic acid or rosmarinic acid in the APTT reagent is 0.5-15mM; the concentration of the salvianolic acid or rosmarinic acid in the PT reagent is 0.05-5mM; and the concentration of the salvianolic acid or rosmarinic acid in the TT reagent is 0.01-2mM. The salvianolic acid or rosmarinic acid can reduce destroys or damages of harmful metal ions and free radicals to tissue factors or thrombin in raw materials or in the process, and greatly improves the oxidation resistance of the coagulative reagent. The antioxidant can be added to other general antioxidants, such as butylated hydroxytoluene or butylated hydroxyanisole, ascorbic acid, glycine and methionine, and the antioxidants are commonly used to further improve the stability of the coagulative reagent.

Description

technical field [0001] The present invention relates to antioxidants for hemostasis and thrombus detection reagents. Background technique [0002] Coagulation testing belongs to the examination of thrombotic diseases and involves several test reagents such as APTT (activated partial thromboplastin time), PT (prothrombin time), TT (thrombin time) and FIB (fibrinogen). [0003] APTT reagent contains phospholipids with distributed unsaturated fatty acid chains, and the double bonds on the fatty acid chains are easily oxidized to form peroxides, malondialdehyde, fatty acids and lysolecithin. Antioxidants are added to reduce the oxidation of liposomes. [0004] PT reagents contain tissue factor (TF) and phospholipids. Tissue factor is a transmembrane single-chain glycoprotein composed of 263 amino acid residues, including the outer, transmembrane and inner membrane regions of the cell membrane. TF initiates the blood coagulation cascade by binding to factor VII / VIIa, which eve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 王华胡小伟高军黄翔宇
Owner QINGDAO GUGAO BIOTECH CO LTD
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