A method for measuring the biological potency of heparin drugs

Abstract

The invention provides a method for determining the biological titer of a heparin drug, and belongs to the field of biological analysis. The method comprises the following steps: mixing a clotting factor with a specific substrate corresponding to a drug to be tested, antithrombin III and a coagulation factor to construct a titer system for an anticoagulant factor in vitro; determining the contentof a reaction product in the titer system by a high performance liquid chromatography-mass spectrometry combined analysis method; calculating the biological titer of the drug to be tested according toa quantitative response parallel line method using a heparin drug of known titer as a standard. The determining method can effectively reduce interference, has strong specificity, is simple in operation, saves time, and can effectively improve the determining efficiency.

Description

technical field [0001] The invention relates to the field of biological analysis, in particular to a method for measuring the biological potency of heparin drugs. Background technique [0002] Blood coagulation consists of a series of chained complex chemical reactions, which constitute the blood coagulation cascade system. At present, there are 12 kinds of blood coagulation factors, which are uniformly marked as Roman numerals I, II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII in the order of discovery in the world. Each chain reaction is a proteolytic reaction, which will convert a zymogen into the corresponding serine protease and finally generate thrombin (FIIa, where a indicates the active form of coagulation factor). Anticoagulants are mostly inhibitors of key enzymes in the coagulation cascade, and are clinically used to prevent and treat thromboembolic diseases. [0003] The most widely used anticoagulants are heparins, including unfractionated heparin (UFH) and lo...

AI Technical Summary

Problems solved by technology

The coagulation method is easily affected by changes in conditions such as laboratories, reagents, and equipment, and has poor reproducibility and low sensitivity. Usually, only semi-quantitative results can be obtained; Western blotting is based on the reaction with several unique anti-heparin antibodies. method, the application is limited

For the chromogenic method: the UV spectrophotometric method used in the current Chinese Pharmacopoeia (ChP2015) requires a large amount of reagents and is expensive, the test cost is high, and it will be interfered by the sample matrix

The fluorescence method has the advantages of strong specificity and high sensitivity, but the background fluorescence of the fluorescent substrate will cause obvious interference to the detection

In addition, the fluorescence method uses human plasma as the enzyme source to establish an enzyme reaction system, which is easily interfered by individual differences in plasma, and the provision and storage of human plasma are also subject to harsh conditions.

Images