Novel method for detecting titer of heparin sample by instrument

A heparin and potency technology, applied in the field of analysis of multi-component biochemical raw materials, can solve problems such as high personnel requirements, cumbersome methods, and improper operation control

Pending Publication Date: 2021-12-17
YANTAI DONGCHENG PHARMA GRP
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

EP7.0 uses the coagulation method to measure heparin potency, which is cumbersome and has poor accuracy; EP8.0 uses the chromogenic substrate method for determination, which simplifies the operation procedure and improves the accuracy, but this method is costly and has a great impact on personnel. The technical requirements are relatively high. At present, the titer detection of heparin samples recorded in EP and USP adopts the chromogenic substrate method. ChP2020 versi

Method used

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  • Novel method for detecting titer of heparin sample by instrument
  • Novel method for detecting titer of heparin sample by instrument
  • Novel method for detecting titer of heparin sample by instrument

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Embodiment Construction

[0020] 1. Instruments and reagents

[0021] Fully automatic sample loading workstation, microplate reader, pipette, pH8.4 buffer, antithrombin, factor Xa, chromogenic substrate S-2765.

[0022] 2. Solution Preparation

[0023] 1) pH8.4 buffer solution: Weigh 6.06g of tris(hydroxymethyl)aminomethane, 2.8g of disodium edetate, 10.23g of sodium chloride, and 1.0g of polyethylene glycol 6000 and dissolve them in 800ml of water. / L hydrochloric acid to adjust the pH value to 8.4 and dilute to 1000ml.

[0024] 2) Antithrombin R1 solution: Add 10ml of pH 8.4 buffer to each bottle of antithrombin to obtain a solution containing 1.0IU / ml of antithrombin, and store it in a refrigerator at 2-8°C.

[0025] 3) Factor Xa solution: Add 12ml of pH8.4 buffer solution to each bottle to dilute to obtain 5.9nkat / ml factor Xa solution. Store in the refrigerator at 2-8°C.

[0026] 4) Chromogenic substrate S-2765 solution: add 11.66ml of water to each bottle of chromogenic substrate S-2765 reage...

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Abstract

The invention discloses a method for detecting the titer of a heparin sample, and relates to an automatic liquid treatment workstation, wherein automation of an experimental process and simulation of a manual detection process can be achieved by utilizing a powerful programming function. The method comprises the following steps: diluting a sample gradient, transferring a sample and adding the sample into an elisa plate, adding a reaction reagent, carrying out heat preserving oscillating, grabbing the elisa plate to a microplate reader, reading, automatically collecting a waste elisa plate, and the like. According to the detection method disclosed by the invention, table tops can be freely integrated according to experimental requirements, a visual editor can complete writing of various programs, the method has a heat preservation oscillation function and can be connected with a plate reading program to realize free transportation and integration of the elisa plate, the detection is not limited by time, and unattended plate arrangement detection can be realized; and the related method can save manpower and reagents, is accurate in detection, good in reproducibility and high in efficiency, and can make up the defects of large error and instability of a manual detection method.

Description

technical field [0001] The invention relates to a method for detecting the titer of a heparin sample, and belongs to the technical field of analysis of multi-component biochemical raw materials. Background technique [0002] The heparin samples in the present invention include heparin sodium / calcium and low molecular weight heparin sodium / calcium. EP7.0 uses the coagulation method to measure heparin potency, which is cumbersome and has poor accuracy; EP8.0 uses the chromogenic substrate method for determination, which simplifies the operation procedure and improves the accuracy, but this method is costly and has a great impact on personnel. The technical requirements are relatively high. At present, the titer detection of heparin samples recorded in EP and USP adopts the chromogenic substrate method. ChP2020 version optimizes the chromogenic substrate method, expands the selection range of reagent volume, and uses the The method changed from a semi-micro reaction to a micro...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N35/00
CPCG01N33/54306G01N35/0099G01N2400/40G01N2035/00465
Inventor 王东玲韩飞吴晓明李颖颖
Owner YANTAI DONGCHENG PHARMA GRP
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