Detection method of cysteine ​​hydrochloride bacterial endotoxin

Abstract

The invention discloses a detection method for cysteine hydrochloride bacterial endotoxin. The method comprises the steps of: a. (I) weighing cysteine hydrochloride and sodium carbonate according to a mass ratio of 20:15-20:17 and performing mixing, adding bacterial endotoxin test water to make the concentration of the test article cysteine hydrochloride in the mixed solution at 100mg / ml; (II) diluting the test article cysteine hydrochloride to 1.25mg / ml-5mg / ml with bacterial endotoxin test water; b. taking a dynamic colorimetric method limulus amoebocyte lysate reagent, opening the reagent, then adding bacterial endotoxin test water to perform re-dissolution; and c. mixing 1.25mg / ml-5mg / ml test article cysteine hydrochloride with the re-dissolved limulus amoebocyte lysate reagent in step b at an equal volume, then putting the mixture into a reaction test tube and inserting the test tube into a dynamic test tube detector to calculate the content of endotoxin. The method provided by the invention avoids the disadvantages of animal pyrogen test methods, and has the advantages of strong repeatability, high sensitivity and simple operation.

Description

technical field [0001] The invention relates to a biological substance of a limulus lysate, in particular to a method for detecting bacterial endotoxin of cysteine ​​hydrochloride. Background technique [0002] Cysteine ​​(Cys) is a common amino acid in living organisms. It is one of the sulfur-containing α-amino acids. It exists in many proteins and glutathione. It can be converted from methionine in the body and can be combined with cysteine. Amino acids are interconverted. Cysteine ​​is stable to acid, but is easily oxidized to cystine by air in neutral and alkaline solutions, and trace iron and heavy metals can promote its oxidation. [0003] Cysteine ​​hydrochloride is an amino acid drug, white crystal or crystalline powder, smelly and sour. Cysteine ​​hydrochloride is easily soluble in water, very slightly soluble in ethanol, and almost insoluble in acetone. Cysteine ​​hydrochloride has reducing properties, can prevent non-enzymatic browning and anti-oxidation, can ...

AI Technical Summary

Problems solved by technology

The gel method is also a method stipulated in the Pharmacopoeia, and the requirements for experimental equipment are low, but this method is a limit determination method. This method can only analyze whether the content of endotoxin in the product exceeds the standard, but cannot obtain exact quantitative results.

The dynamic turbidity method is the most commonly used quantitative determination method, which is much more sensitive than the gel method, but it cannot obtain satisfactory results in terms of sensitivity, detection range, linearity, anti-interference ability, and reliability of detection results. The consumption of Limulus reagent is large, which is not conducive to the protection and sustainable development of Limulus

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