Research On Quality Control Technology Of Amino Acid And Peptide Drugs

Biochemical drugs are biochemical basic substances used for the prevention,treatment,and diagnosis of diseases,which are obtained from the separation and purification of living organisms,chemical synthesis,microbial synthesis or modern biotechnology.Amino acids and polypeptides drugs are an important part of biochemical drugs.This paper focuses on the research of quality control technology of amino acids and polypeptide drugs,and studies the main components,excipients and buffer components in various amino acids and polypeptides drugs.In the first part of the research,the determinations of the main components of amino acids and polypeptides were established,and the potassium aspartate and magnesium aspartate injection,spleen amino peptide oral solution and armillariella oral solution were taken as the subjects of the study.Firstly,the optimized chromatographic condition for the determination of the main components(aspartate,potassium ion,magnesium ion)and the impurity ion(sodium ion)in potassium aspartate and magnesium aspartate injection was established.Through single-factor analysis,the effects of column temperature,initial organic phase ratio,buffer types,concentration,and pH on chromatographic separation were investigated,and the retention mechanism of mixed mode chromatography was explained.Based on the single-factor experiments,four-factor and three-level response surface experiments were designed to obtain optimal chromatographic conditions.The optimized method has the advantages of high sensitivity,good repeatability,accuracy,robustness,and short time-consumingsaving time.It can be used as an alternative to ion chromatography and high performance liquid chromatography for the quality control of potassium aspartate and magnesium aspartate injection.Secondly,the determination of free amino acids and total amino acids in the fungal extracts(armillariella oral solution)and animal spleen extracts(spleen amino peptide oral solution)were studied.The chromatographic methods of determination of the free amino acids and total amino acids in two types of multi-component amino acids and polypeptides drugs were established.The different contents of the total amino acids and the free amino acids represent the contents of polypeptides or proteins.The method realized the simultaneous determination of the main components of the free amino acids,proteins and polypeptides.The established method overcome the disadvantages of colorimetric methods,such as the existing of multiplex interfering substances and showing poor specificity when measuring the contents of polypeptides and proteins.The pre-column online automatic derivatization method was simple,and showed a good repeatability.This method can be used for the quality research of other types of multi-component biochemical drugs and provides references for the study of the mechanism of action of complex multi-component amino acids and polypeptides drugs.In the second part of the research,the excipients and buffers in amino acids and peptides drugs were studied.The compound amino acid(15)dipeptide(2)injection and the recombinant anti-EGFR human/mouse-chimeric monoclonal antibody for injection were taken as the research objects.Firstly,the methods of the determination of free acids(acetic acid,citric acid),free ammonia and sodium ion in compound amino acid(15)dipeptide(2)injection were studied.Various analytical techniques were investigated.After optimization of chromatographic conditions,an ion exclusion chromatography method was respectively established to determine acetic acid,an anion exchange chromatography method to determine citric acid and a cation exchange chromatography method to determine free ammonia and sodium ion.All the three analytical methods were validated by methodology.Accuracy,sensitivity,repeatability and linear range all meet the testing requirements.Using acidic,alkaline aqueous mobile phase,without adding any organic reagents,so these methods are nontoxic and environment-friendly.The determination of free acids,free ammonia and other ionic components in injections allows speculation of the formulation process for injections and facilitates the development of consistency assessment for injections.Secondly,the determination of sucrose and polysorbate 80 in recombinant anti-EGFR human/mouse-chimeric monoclonal antibody for injection were established,respectively.The sample pretreatment method was optimized,and a heating denaturation method was established to overcome the instability of sucrose in sulfosalicylic acid.The methods for determining excipients in injections helps in controling the quality of drugs and ensuring the safety of the drugs while keeping effective.In summary,the research of this dissertation established methods for the determination of the main components,excipients and buffering components in various amino acids and polypeptides drugs by means of various analysis methods and analysis techniques,which contributes to the improvement of the quality standards of these drugs and the development of the consistency evaluation of drug.