| C1 is a new drug that has not been marketed at China and abroad.According to Chinese chemical drug registration classification standards,it belongs to category 1.1new drugs.There are no quality standards for C1 APIs and their preparations.In order to ensure the safety and effectiveness of the drugs,this project conducts a comprehensive and systematic quality study of C1 APIs and tablets to provide experimental basis for C1 declaration.The main contents of this research are as follows:一.C1 API quality research: 1.Focus on the inspection of C1 related substances:use high performance liquid chromatography to detect C1 related substances,and analyze the structure of three impurities(I,III,IV)whose content exceeds 0.1% by area normalization method.Among them,impurity Ⅲ is destroyed and degraded by high temperature of the main drug C1,and then separated and prepared by liquid chromatography to obtain pure product.The purity analysis by HPLC is 97%.The pure product of impurity Ⅲ is analyzed by UV,MS,IR and NMR.Impurity III is C1monoester;Impurity I is analyzed by liquid chromatography retention time and UV spectroscopy,which is consistent with the chromatographic behavior of C1 starting material drug.It is preliminarily inferred that impurity I is C1 starting material drug,and further confirmed by HPLC-MS analysis The molecular weight of impurity IV is28 higher than that of C1.It is preliminarily inferred that a carboxyl group in C1 is esterified by ethanol,that is,the C1 diester is then esterified to become a triester,so impurity III is a triester.,the detection limit and quantification limit of impurity I are respectively 100 ng and 200 ng,the detection limit and quantification limit of impurityⅢ are 20 ng and 100 ng respectively;2.C1 identification: C1 is identified by color reaction,infrared spectroscopy,and liquid chromatography;3.C1 inorganic impurity inspection: C1 chloride is less than 0.005%,sulfate is less than 0.05%,Heavy metals are less than 20 ppm,residue on ignition is less than 0.2%,loss on drying is less than0.5%,arsenic salt is less than 2 ppm;4.Residual solvent inspection: for the three organic solvents of ethanol,acetonitrile and pyridine used in the synthesis and purification of C1,establish Headspace gas chromatography,with methanol as the internal standard,was used to detect the residual amount of three organic solvents.Under the established chromatographic conditions,the recovery rates of the three organic solvents are between 98% ～ 102%(RSD<2.0%),and the repeatability is good,which can meet the C1 residual solvent detection;5.Assay of C1 API: establish a high performance liquid chromatography method to determine the content of C1,mobile phase: A(acetonitrile /Isopropanol 4:1,V/V)-B(20 m M KH2PO4,adjust the p H to 2.0 with H3PO4);gradient elution:(0～15 min 2%-40%A;15～20 min 40%-2%A;20～30 min2%A);flow rate 0.8 ml/min;detection wavelength 210 nm.Methodological verification shows that the method has good repeatability and precision,and the detection limit and quantification limit of C1 are 40 ng and 80 ng,respectively.The HPLC-MS method and volumetric analysis method are used to support the high performance liquid chromatography.The results show that the liquid chromatography method is accurate and easy to operate.Therefore,it is used as a routine method for the determination of C1 content;6.The influence factor test of C1API: C1 has no change in properties under high temperature,high humidity and strong light damage conditions;the content and related substances are basically unchanged,indicating that C1 is stable under high temperature,high humidity and light conditions.two.C1 tablet quality research: 1.C1 tablet content determination: chromatographic conditions adopt the C1 bulk drug content determination method,method verification shows that C1 tablet excipients do not affect the tablet content determination,the method precision and repeatability are good,and the sample is added The recovery rate was 100.36%(n=9,RSD<2.0%),indicating that this method is suitable for the determination of C1 tablet content.2.Determination of related substances in C1 tablets:The system suitability test shows that the method is good,and the specificity test shows that C1 tablets are stable under high temperature,high humidity,and light conditions,and the impurity Ⅲ content increases under acid,alkali,and oxidative damage conditions;3.C1 tablet dissolution test: the paddle method was used,with 0.1mol/L hydrochloric acid as the dissolution medium,and the rotating speed was 50 rpm,to conduct the C1 tablet dissolution test.The dissolution curve shows that the dissolution rate of C1 reaches more than 90% in 10 min,and the dissolution is good.This subject conducts quality research on C1 bulk drugs and their tablets.The above-mentioned experimental research provides experimental basis for the formulation of C1 quality standards and provides guarantee for the quality control of C1. |
