| Chondroitin sulfate(CS)is extracted and purified from cartilaginous tissues which are composed of alternate sequences of D-glucuronic acid and N-acetyl-D-galactosamine.Disaccharides with different numbers and positions of sulfate groups can be located inside the polysaccharide chains.Chondroitin sulfate has been demonstrated to possess varieties of pharmacological activities,including anti-inflammation,anti-tumor,anti-atherosclerosis and anti-hepatitis.Due to the source variety,structure complexity and extensive application of chondroitin sulfate,it is urgent to improve its quality control.ChP,EP,USP and BP collected CS active pharmaceutical ingredient(API)for oral,while CS injection,specific variety in China,was collected by National Drug Standards.Injection has a higher security risk than oral preparation.On the contrary,CS injection has a lower quality specification than oral preparation.It is necessary to improve quality specification of CS,especially CS injection.This paper is divided into four sections.The first section is about sources.As is described in the quality specification,CS is obtained from cartilages of porcine,such as larynx bone,nasal septum and trachea,in ChP.Blending other sources will introduce risk for injection.As a consequence,this part is about to establish a possible more specific analysis method to distinguish between various origin sources according to the study of specific optical rotation and disaccharide analysis.CS from different animal sources exhibits a wide variety of disaccharide composition.Particularly,4S / 6S is an important indicator of different sources of chondroitin sulfate.The second section is about the method for injection assay.It is important for accuracy content to assure effectiveness and production process stability of CS.The Elson-Morgan method for injection in current standard has poor accuracy,sensitivity,complexity and repeatability.With reference to content determination method for oral API of ChP,this part established enzymatic HPLC for injection API and accomplished validation of this method.Compared with the original method,thismethod has the advantages of good accuracy,simple operation and good repeatability.The third section is about impurities.CS is extracted and purified from the pig cartilage and there may be some other impurities residual,such as polysaccharides,organic solvents and proteins.This paper built up for the first time analysis methods for relative substance analysis,which were cellulose acetate membrane electrophoresis and HPLC method,and compared the advantages and disadvantages of the two methods.With reference to residual solvents determination method for oral API of ChP,this paper set up gas chromatography analysis method for injection API and accomplished validation of this method.The residual protein of CS for injection was measured by Lowry method.The last section is about the molecular weight and molecular weight distribution.The SEC-HPLC method in current standard whose reference substance is not reasonable,is not accurate comparing to SEC-MALLS method.To adopt SEC-HPLC method to determine the molecular weight and molecular weight distribution,this paper optimized the SEC-HPLC method,and the method was used for the screening CS molecular weight calibration reference standard.In conclusion,this paper established methods including the source analysis,content determination,impurity analysis and molecular weight and molecular weight distribution,to improve the quality control of chondroitin sulfate.This research contributes to improvement of quality standard for chondroitin sulfate. |
