Novel ellagic acid compound and method for preparing activated partial thromboplastin time measuring reagent through ellagic acid compound

A technology of thromboplastin time and ellagic acid, which is used in biological testing, material testing products, etc., can solve the problems such as the inability to guarantee the active stability of the product, the proportion of phospholipids, affecting the test results and reagent stability, and prolonging the APTT measurement time. , to achieve the effects of improving solubility, reducing batch-to-batch difference, and optimizing components

Inactive Publication Date: 2019-04-12
太原博奥特生物技术有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

The application of APTT to the determination of the coagulation system is a simple, fast and cheap project. At present, the main problem of using ellagic acid as an activator is the low solubility of ellagic acid in water, and the formation of complexes between ellagic acid and metal ions Only then can the endogenous coagulation system be activated. When ellagic acid forms an activator with metal ions, it is easier to form a precipitate, which reduces the effective ellagic acid component in the solution, prolongs the APTT measurement time, and affects the test results and the stability of the reagent.
Another problem is that it is difficult to standardize APTT reagents at home and abroad. The main reason is that the types of phospholipids are different. The phospholipids contained in APTT reagents currently on the market are cephalin extracted from animals. The phospholipids from this source cannot guarantee the stability of product activity. The nature and proportion of phospholipids extracted

Method used

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  • Novel ellagic acid compound and method for preparing activated partial thromboplastin time measuring reagent through ellagic acid compound
  • Novel ellagic acid compound and method for preparing activated partial thromboplastin time measuring reagent through ellagic acid compound
  • Novel ellagic acid compound and method for preparing activated partial thromboplastin time measuring reagent through ellagic acid compound

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Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1: A novel ellagic acid compound is prepared by mixing ellagic acid and a solid dispersant at a mass ratio of 1:20, and the solid dispersant is polyvinylpyrrolidone PVPk30; its preparation method is: precision weighing Take ellagic acid and dispersant, mix them, dissolve them evenly in methanol to a final concentration of 30mg / ml, stir and dissolve at room temperature, evaporate at 33kPa at 40°C until the methanol is completely removed, and dry and store.

[0022] The activated partial thromboplastin time assay reagent prepared by utilizing the described ellagic acid compound, the reagent is configured from raw materials with the following final concentration: liposome 20 µg / ml, ellagic acid compound 0.3 mg / ml, metal Ion 20mM, stabilizer 1% W / V, preservative 0.1% W / V; the preparation method is: weigh the ellagic acid compound in proportion and dissolve it in HEPES buffer, stir at room temperature for 2h to mix, add preservative, stabilize Agent, mix well, add...

Embodiment 2

[0025] Embodiment 2: A kind of novel ellagic acid compound is prepared by mixing ellagic acid and a solid dispersant according to a mass ratio of 1:9; the solid dispersant is cellulose acetate butyrate CMCAB; its preparation method is: Accurately weigh ellagic acid and dispersant, mix them, and evenly dissolve them in an organic solvent with a volume ratio of acetone and ethanol of 1:4 to make a final concentration of 20mg / ml, stir and dissolve at room temperature, evaporate at 80kPa-40kPa, and remove the organic solvent at 40°C. Solvents can be stored dry.

[0026] The activated partial thromboplastin time assay reagent prepared by using the described ellagic acid compound is configured from raw materials with the following final concentration: liposome 40ug / ml, ellagic acid compound 0.6mg / ml, metal Ion 20mM, Stabilizer 1% W / V, Preservative 0.1% W / V. The surfactant is 3-[3-(cholamidopropyl)dimethylamino]propanesulfonic acid inner salt CHAPS. The metal ion is a divalent meta...

Embodiment 3

[0027] Embodiment 3: A novel ellagic acid compound, prepared by mixing ellagic acid and a solid dispersant at a mass ratio of 1:14; the solid dispersant is hydroxypropyl cellulose phthalate HPMCP; its The preparation method is: accurately weigh ellagic acid and dispersant, mix and dissolve in an organic solvent with a volume ratio of acetone and ethanol of 1:4, so that the final concentration is 20mg / ml, stir and dissolve at room temperature, evaporate at 80kPa-40kPa, and evaporate at 40 degrees Remove the organic solvent and store in a dry place.

[0028] The activated partial thromboplastin time assay reagent prepared by using the described ellagic acid compound is configured from raw materials with the following final concentration: liposome 100ug / ml, ellagic acid compound 0.6mg / ml, metal Ion 30mM, Stabilizer 1% W / V, Preservative 0.1% W / V. The surfactant is octyl-β-D-glucopyranoside OGP. The metal ion is a divalent metal ion CuCl 2 , other methods are the same as the met...

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Abstract

The invention belongs to the technical field of blood coagulation reagent preparation, and provides a novel ellagic acid compound aiming at the characteristic that ellagic acid is difficult to dissolve in water. The novel ellagic acid compound is prepared from the ellagic acid and a solid dispersant which are mixed according to a certain proportion. The solubility of the ellagic acid is improved,the problem that the ellagic acid is prone to being precipitated is solved, and the stability of the ellagic acid in products is guaranteed. The invention also provides a method for preparing an activated partial thromboplastin time measuring reagent through the ellagic acid compound. The reagent is prepared from liposomes, the ellagic acid compound, metal ions, a stabilizer and a preservative according to a certain proportion. According to the prepared reagent, through the ellagic acid solid compound, the problem that the ellagic acid is prone to being precipitated is solved, components of the liposomes are optimized, the inter assay variation between the activated partial prothrombin reagent products is lowered, the stability of the products is good, and the quality of the activated partial prothrombin reagent is guaranteed.

Description

technical field [0001] The invention belongs to the technical field of blood coagulation reagent preparation, and in particular relates to a novel ellagic acid compound and an activated partial thromboplastin time determination reagent prepared by using the ellagic acid compound. Background technique [0002] Activated partial thromboplastin time APTT (activated partial thromboplastin time) is a more sensitive test commonly used to screen whether the endogenous coagulation system is normal. It is widely used in the initial screening diagnosis of bleeding diseases. Testing, laboratory testing of heparin anticoagulant quality, testing of lupus anticoagulant substances, etc. [0003] The principle of activated partial thromboplastin time (APTT) is to add the plasma to be tested into the APTT reagent, mix it at 37 degrees and preheat it for a certain period of time, use the activator to activate FXII, use phospholipids as part of the thromboplastin, and observe the plasma in the...

Claims

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Application Information

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IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 赵亚蕊赵邑赵峰梅李红霞潘薇薇张全爱曹鹏程
Owner 太原博奥特生物技术有限公司
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