Method for detecting macromolecular substances in Shengqi Fuzheng injection

Abstract

The invention relates to a method for detecting macromolecular substances in a Shengqi Fuzheng injection. The method includes steps of firstly, performing ultrafiltration centrifugal treatment to the Shengqi Fuzheng injection using an ultrafiltration centrifugal method, adding ultrapure water to upper trapped fluid obtained by the ultrafiltration centrifugal treatment, mixing well and centrifuging until a washed out part is negative in a Molish reaction detection, and collecting the upper trapped fluid; secondly, using a BCA (bicinchoninic acid) method and the Molish method to detect the upper trapped fluid obtained by the first step so as to determine whether the macromolecular substances exist in the Shengqi Fuzheng injection or not. The method is simple and common in instruments and materials and in experimental method, low in cost, convenient to use in practical production, provides reference of quality control during manufacturing process of a manufacturer, and further guarantees safety of the Shengqi Fuzheng injection.

Description

technical field [0001] The invention belongs to the technical field of drug analysis, in particular to a method for detecting macromolecular substances in Shenqi Fuzheng injection. Background technique [0002] As a traditional Chinese medicine injection, Shenqi Fuzheng Injection is mainly prepared from Codonopsis pilosula and Radix Astragali. During the extraction process of the raw materials of Shenqi Fuzheng Injection, a large amount of macromolecular impurities such as protein, polysaccharide, polypeptide, and tannin will be produced. Macromolecular impurities are the main cause of adverse reactions such as allergic reactions. Among them, macromolecular polysaccharides are generally proteoglycans, and the antigenicity of this glycoprotein is higher than that of the original protein. Enhances allergic reactions. Therefore, if these macromolecules cannot be removed during the production of injections, they may cause adverse reactions such as allergies and redness in patie...

AI Technical Summary

Problems solved by technology

[0004] At present, the academic community has not formed a unified limit for the molecular weight of macromolecular substances that cause adverse reactions. It is believed that the curative effect of traditional Chinese medicine injections comes from the small molecular substances contained in them, and has nothing to do with substances with a molecular weight greater than 3000. However, polysaccharides with a molecular weight greater than 3000 in many traditional Chinese medicines The macromolecular substances of protein and protein still have strong medicinal activity. If substances with a molecular weight greater than 3000 are removed from such traditional Chinese medicine injections, the safety of the injection will increase, but the efficacy will be greatly reduced.

At present, there is no document disclosing the specific molecular weight limit of macromolecular substances that are prone to side reactions in Shenqi injection, and conventional polysaccharide and protein detection methods cannot detect macromolecular substances with different molecular weights. For Shenqi Fuzheng injection , there is no relevant literature report on the detection method that can not only ensure its safety but also ensure its effectiveness

[0005] Shenqi Fuzheng Injection is different from ordinary injections in that, due to the inevitable pigment residues in the extraction process of traditional Chinese medicine, direct use of chromogenic reactions such as Folin-phenol reagent method and Bradford method to detect proteins and polysaccharides will lead to detection The result produces serious errors. Therefore, before using the chromogenic method for detection, it is necessary to remove the pigment in the injection. For example, Chinese patent application 201010504574.4 discloses a method for detecting trace proteins in Chinese medicine injections. However, when the Chinese medicine injection itself has pigments (Shenqi Fuzheng Injection is a yellow liquid), it is necessary to increase the decolorization step so as not to affect the final result, and when using a protein-specific adsorption membrane for decolorization, the soaking time needs to be at least 15 hours. It can be seen that the operation time of this method is too long. long, complicated steps

[0006] Shenqi Fuzheng Injection is a traditional Chinese medicine injection. The protein and polysaccharide macromolecular components in the injection cannot be detected by conventional detection methods, because other components may have a greater impact on the detection results when conventional methods are used for detection.

In order to eliminate the influence of other ingredients, the detection methods of macromolecular substances in traditional Chinese medicine injections are usually complicated

Chinese patent ZL200610001838.8 discloses a method for determining the content of polymer substances in injections. This method uses high-performance liquid chromatography. Although the sensitivity of the determination results is high, it needs to use reference substances, and the operation steps are relatively complicated.

[0007] Chinese patent application 201110002873.2 uses enzyme-linked immunosorbent assay (ELISA) to measure trace protein in Shengmai injection, which has many steps and high cost

[0008] The current Pharmacopoeia (2010 edition) has formulated the substance detection standards for proteins, tannins, resins, and oxalates in traditional Chinese medicine injections, but has not specifically formulated quality control standards for macromolecular substances such as proteins, polypeptides, and polysaccharides.

Moreover, the turbidimetric method of sulfosalicylic acid used to control the protein of traditional Chinese medicine injection in the Pharmacopoeia has low sensitivity and is easily affected by the interference of the color of the injection itself.

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