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Ellagic acid reagent and preparation method thereof as well as activated partial thromboplastin time (APTT) determination reagent and APTT kit

A preparation method and technology of ellagic acid, applied in the direction of biological testing, material inspection products, etc., can solve the problems of loss of activity, precipitation, and affecting the quality of domestic reagents, and achieve good stability and simple and fast process

Active Publication Date: 2018-06-29
WUHAN CHANGLI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, as the type of synthetic phospholipids, mixing three types of phosphatidylethanolamine (PE), phosphatidylcholine (PC) and phosphatidylserine (PS) into the reagent has become the mainstream, but there are many types of synthetic phospholipids, and it is not easy to Obtained and expensive, in addition, the proportion of each component is difficult to control, and there are also dissolution problems, some of which will lead to precipitation and affect the quality of reagents
Existing reagents will have ellagic acid and phospholipid precipitation problems, ellagic acid buffer solution, raw material treatment such as ellagic acid will lose activity under alkaline conditions, whether the phospholipid emulsification is uniform, and the ratio of ellagic acid buffer solution to phospholipids etc. have directly affected the quality of domestic reagents, resulting in the market being still dominated by giants such as Siemens, Starco, and Werfen.

Method used

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  • Ellagic acid reagent and preparation method thereof as well as activated partial thromboplastin time (APTT) determination reagent and APTT kit
  • Ellagic acid reagent and preparation method thereof as well as activated partial thromboplastin time (APTT) determination reagent and APTT kit
  • Ellagic acid reagent and preparation method thereof as well as activated partial thromboplastin time (APTT) determination reagent and APTT kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] This embodiment provides an ellagic acid reagent, which is prepared by the following preparation method:

[0046] (1) Add 0.4 mmol ellagic acid into calcium-free purified water, and stir rapidly to make it evenly dispersed.

[0047] 0.5 mmol sodium acetate was added.

[0048] Quickly add 1 mmol Tris-hcl within 10 minutes after adding ellagic acid to adjust the pH to 7.1±0.1.

[0049] Add 0.025mmol iron ion.

[0050] Arginine hydrochloride 1.5% and glycine 1.5% were added.

[0051] Add the preservative sodium azide.

[0052] (2) Preparation of emulsified phospholipids: Add calcium-free purified water at 5°C to 0.32 g / L rabbit cephalin, mix and emulsify in an ice bath at 5°C to obtain emulsified phospholipids.

[0053] (3) Add the emulsified phospholipid into the buffer solution in a specific ratio, stir and mix quickly to obtain a yellow uniform dispersion, that is, the ellagic acid reagent.

[0054] This embodiment also provides a reagent for measuring activated pa...

Embodiment 2

[0056] This embodiment provides an ellagic acid reagent, which is prepared by the following preparation method:

[0057] (1) Add 0.2 mmol of ellagic acid into calcium-free purified water, and stir rapidly to make it evenly dispersed.

[0058] Add 0.4 mmol Tris-base.

[0059] Quickly add 0.8mmol Hepes within 10 minutes after adding ellagic acid to adjust the pH to 7.1±0.1.

[0060] Add 0.02 mmol of magnesium ions.

[0061] Add alanine.

[0062] Add preservative proclin300.

[0063] (2) Preparation of emulsified phospholipids: Add calcium-free purified water at 3°C ​​to 0.3 g / L rabbit cephalin, mix and emulsify in an ice bath at 3°C ​​to obtain emulsified phospholipids.

[0064] (3) Add the emulsified phospholipid into the buffer solution in a specific ratio, stir and mix quickly to obtain a yellow uniform dispersion, that is, the ellagic acid reagent.

[0065] This embodiment also provides a reagent for measuring activated partial thromboplastin time, which includes the ab...

Embodiment 3

[0067] This embodiment provides an ellagic acid reagent, which is prepared by the following preparation method:

[0068] (1) Add 0.5 mmol of ellagic acid into calcium-free purified water, and stir rapidly to make it evenly dispersed.

[0069] 0.8 mmol sodium hydroxide was added.

[0070] Quickly add about 0.5ml of acetic acid within 10 minutes after adding ellagic acid to adjust the pH to 7.1±0.1.

[0071] Add 0.03 mmol manganese ions.

[0072] Add histidine and glutamic acid.

[0073] Add the preservative gentamicin.

[0074] (2) Preparation of emulsified phospholipids: Add calcium-free purified water at 8°C to 0.35 g / L rabbit cephalin, mix and emulsify in an ice bath at 8°C to obtain emulsified phospholipids.

[0075] (3) Add the emulsified phospholipid into the buffer solution in a specific ratio, stir and mix quickly to obtain a yellow uniform dispersion, that is, the ellagic acid reagent.

[0076] This embodiment also provides a reagent for measuring activated partia...

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Abstract

The invention provides an ellagic acid reagent and a preparation method thereof as well as an activated partial thromboplastin time (APTT) determination reagent and an APTT kit and relates to the field of clinical diagnosis reagents. The preparation method comprises the following steps: adding ellagic acid into calcium-free purified water and rapidly stirring to uniformly disperse the ellagic acid; adding an alkaline buffering solution or inorganic alkali; rapidly adding an acidic buffering solution or organic acid and inorganic acid within 10min after the ellagic acid is added; adding metal ions; adding amino acid and a derivative thereof to obtain a buffering solution; furthermore, adding the calcium-free purified water into rabbit brain phospholipid and uniformly mixing and emulsifyingto obtain emulsified phospholipid; adding the emulsified phospholipid into the buffering solution and rapidly stirring and uniformly mixing to obtain the ellagic acid reagent. The technology is simpleand rapid and has detailed descriptions. The activated partial thromboplastin time determination reagent comprises the ellagic acid reagent and a calcium chloride reagent and has good stability. TheAPTT kit containing the activated partial thromboplastin time determination reagent can be used for directly replacing an imported high-stability APTT kit.

Description

technical field [0001] The invention relates to the field of clinical diagnostic reagents, and in particular to an ellagic acid reagent and a preparation method thereof, a reagent for measuring activated partial thromboplastin time, and an APTT kit. Background technique [0002] In the field of clinical examinations, it is known to measure blood coagulation time to study abnormalities of blood coagulation factors, among which APTT measurement is one of general clinical examinations of blood coagulation ability. APTT (activated partial thromboplastin time), namely activated partial thromboplastin time, refers to the clotting time that reflects the function of the intrinsic coagulation pathway that blood starts through contact with negatively charged foreign bodies during bleeding. APTT measurement is used not only for screening tests for the deficiency or abnormality of intrinsic coagulation factors but also for monitoring heparin therapy and the like. [0003] Most of the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 李幼鹏王宇
Owner WUHAN CHANGLI BIOLOGICAL TECH CO LTD
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