The Research Of The Quality Control Of SR-H22

Drug quality control research is an important part of new drug research.Quality control is not only need for drug substances and drug products, but also need to accessories, packaging materials inside and outside,also include the new drug research and development, production, storage and transportation, and other key control.In order to ensure the safety of the drug quality, effective and controllable.The derivatives SR-H22 of Benzene and triazine-1,4-dioxide is developed by our team,the quality standards of SR-H22 were well studied.1.The first part: the research of quality standards of SR-H22The synthetic route of SR-H22 and melting point of various intermediates, UV spectrume, the contents were determined.2.The SR-H22 quality control research(1) The establishment of the content determination method of SR-H22We established the content determination method of SR-H22, the chromatographic conditions are as follow: chromatographic column: Diamond C18 Column(250mm×4.6mm, 5um); The mobile phase is acetonitrile-water(25:75,V/V);Detection wavelength is 243nm; Flow rate: 1.0ml·min-1; Sample concentration:25ug·ml-1; Sample size: 20ul; Column temperature: 35℃. After the construction of the liquid phase method, the sensitivity, precision, repeatability and linear range were investigated, and it indicated that the liquid phase method is stable and reliable, which can be used for the content determination of SR-H22.Under this chromatographic conditions, the theoretical plate number calculated on SR-H22 peak is no less than 7000, separating degree is no less than 1.5. After methodology investigation, this linear relation of peak area and concentrationin is good in range 5ug·ml-1-100ug·ml-1(r=0.9979), linear equation is:Y=1.8105×105X-3.2081×105, The LOD is 1ng and LOQ is 3ng, The days precision and the daytime precision RSD were both less than 2%.(2) The SR-H22 relevant material check method is establishedWe established the relevant material check method of SR-H22, the chromatographic conditions is as follow: chromatographic column: Diamond C18Column(250mm×4.6mm, 5um); The mobile phase is acetonitrile-water(25:75, V/V);Detection wavelength of 243nm; Flow rate: 1.0ml·min-1; Concentration of the sample:100ug·ml-1; Sample volume 20ul; Column temperature: 35℃.The affect of Acid, alkali, oxidation, high temperature, strong light on SR-H22 APIs were inspeted. The experimental results showed that: When the SR-H22 APIs in1mol·L-1HCL, 30% H2O2, glare, 105℃ oven for 12 h, There were no significant degradation products found; They show the stablebility of SR-H22 APIs in acid,oxidation, light and high temperature condition.But in the environment of1mol·L-1Na OH alkaline,room temperature placed 1h,we can found the obvious degradation products in SR-H22 APIs, the main peak purity reduced to 80%; If we heated the SR-H22 APIs, it will find main completely disappear instead of degradation products. We can use the former chromatography method to make main peak and degradation products complete separation.We use the HPLC-MS/MS to analysis the degradation products of SR-H22, we use the same chromatographic conditionsas former, the analysis mode is SIM mode which detect the related substances 2 sample solution,the m/z of material 2 is 163,when the Fragmentor is 110 V, m/z 163 positive ion peak response is best. Similarly,the related substances 1 which m/z is 147 take the same way of experimental investigation, found that when the Fragmentor is 90 V, positive ion peak response is the 147 best.Examine Collision Energy: when we need to detect m/z 163, Fragmentor is 110 V,the Collision Energy is set to 14 V, 22 V, 30 V, 42 V. Under different collision energy,m/z for 39, 44, 65, 80, 91, 133, 163 pieces of peak has appeared, they are the product ion structure of m/z 163. When the detect ion m/z 147, Fragmentor is 90 v, The Collision Energy is set to 18 V, 22 V, 26 V, 46 V. Under different collision energy, m/z to39, 65, 91, 65 pieces of the peak has appeared, they are the product ion structure of m/z 147. We can determine that the relevant material is intermediates SR-H02 by this information.(3) The determination of residual solvent of SR-H22In order to ensure the safety of the drug,we use the gas chromatography to determine the organic solvent residue in the samples, method we established the methods of four kinds residual solvent in SR-H22 by capillary gas chromatography,The methods are as follows: chromatography column: DB-WAX(30m×0.45 mm,0.45um) capillary chromatographic column; The carrier gas: high purity nitrogen;Detector: FID detector. Using temperature programmed, column temperature, initial temperature of 35℃, 7min, and then to rise to 200℃, 35℃/min rate keep 6min,column head pressure: 4psi. Injection port temperature: 220℃, detector temperature:250℃. Direct shunt, DMSO as the solvent, determine the content of residual solvents.After the establishment of Methods. We investigate the sensitivity, precision,repeatability and linear of this method.The result is well.(4) The stability testThe factors experimentThe SR-H22 APIs were placed in high temperature(60℃)、 high humidity(92%+5%) and glare(4500 lx+500lx) under the condition of opening placed 10 days,in 5 and 10 days sampling test, we inspected the stability of SR-H22 APIs, tried to learn about the the factors influencing the stability of SR-H22 and the possible degradation pathway and degradation products.Experimental results showed that under the condition of high temperature, high humidity and strong light irradiation test stable place for 10 days, there were no significant degradation appeared in SR-H22 APIs, the character,content and related substances of SR-H22 are not seen obvious changes.(5)The chromatography(HPLC) analysis method of starting material SR-H01,intermediates SR-H02.The liquid phase analysis method of SR-H01 are as follows: chromatography column Dikma Diamonsil C18(250mm×4.6mm,5um) mobile phase: acetonitrile-water(45:55, V/V), detection wavelength of 225 nm, velocity: 1.0 ml·min-1, column temperature 35℃, sample volume: 20 ul. The methodology in the liquid phase method,the SR-H01 detection limit(LOD) for 2ng, quantitative limit(LQD) for 6ng; the linear range is good in 10ug·ml-1-150ug·ml-1 range, the method of precision, days precision are all less than 2%.The SR-H02 liquid phase analysis method is the same with SR-H22:chromatography column Dikma Diamonsil C18(250mm×4.6mm,5um) mobile phase:acetonitrile-water(25:75, V/V), detection wavelength of 238 nm, velocity: 1.0 ml·min-1, column temperature 35℃, sample volume: 20 ul. The methodology in the liquid phase method, the SR-H02 detection limit(LOD) is 1ng, quantitative limit(LQD) is3ng; The linear range is well in 5ug·ml-1-100ug·ml-1 scope, precision, days precision are all less than 2%.(6)The physical and chemical properties and inspectionBy thin layer chromatography, ultraviolet spectrophotometry and high performance liquid chromatography(HPLC) method,we can identify the sample and reference substance. According to the Chinese Pharmacopoeia 2015 edition of the fourth general requirements, the three batch samples(batch number: 20151106,20151109, 20151113) to the physical and chemical properties(properties, solubility,p H value, melting point, etc.), inorganic inspection. This product is a yellow powder,odorless.The samples are slightly dissolved in In methanol, acetonitrile, ethanol, N,N-dimethyl formamide, slightly soluble in water, 0.1mol·L-1 hydrochloric acid,0.1mol·L-1sodium hydroxide. The melting point is 212.3℃ to 213.5℃, There are no wet in sample. Inorganic inspection results show that the Chloride< 0.01%,Sulfate<0.04%, Dry weight loss<20 PPM, Arsenic salt<2PPM. Ignition residue<0.2%.(7)SummaryReference to <Chinese Pharmacopoeia>,<chemical drug quality standard technical guiding principles>; we tried to establish the standardization of the SR-H22APIs; we also verify the specificity of this method and we inspect the two related substances in alkali destroy the sample; we also inspect the residual solvent of SR-H22 and the factors testings were done., they will be benefit for the further research of the1,2,4-benzotriazine-1,4-dioxide derivatives of SR-H22 by provides experimental data.