Studies On Drug Quality Control Of Dronedarone Hydrochloride And The Film-coated Tablet

Atrial fibrillation is one of the most common arrhythmia in clinical, andthe incidence increased with age. Atrial fibrillation seriously affected not onlythe quality of life in patients, but also leaded to arterial embolism, stroke,myocardial infarction, and other high-risk diseases. Although radiofrequencycatheter ablation was one of methods for the treatment of atrial fibrillation, itwas affected by the high cost of treatment and treatment technologies withhigher limits. The most patients who received radiofrequency catheter ablationstill needed long-term treatment with drugs to control the onset of atrialfibrillation. The drug treatment of atrial fibrillation was still the main methodat the present stage.Dronedarone hydrochloride was a new antiarrhythmic drug, its chemicalstructure was similar to amiodarone, the standard drug of treatment of atrialfibrillation. The mechanism of dronedarone hydrochloride was the same asamiodarone’s. Both of them were potassium channel blockers. Dronedaronedid not contain iodine, with weak lipophilic than amiodarone, and had lessside effects than amiodarone outside of cardiovascular system. Dronedaronehad little effect on thyroid hormone receptors, and had neither significantcardiac toxicity, nor torsade de pointes. The tolerability of the new drug wasgood in clinical. The result of phase III clinical trial confirmed thatdronedarone hydrochloride was effective in reduction of the recurrence ofatrial fibrillation or atrial flutter, and the morbidity and mortality ofcardiovascular diseases. The new product is not marked in China and theresearch on the qulity control was not found in domestic. This paper willgreatly contribut to the self-made dronedarone for development, productionand clinic usage in our country. Part one Studies on quality control of dronedarone hydrochlorideObjective: To study physical and chemical properties of dronedaronehydrochloride which relate to its quality, to establish new methods and offerdatas for quality control.Methods:1. Related substances: The HPLC method was adopted todetect the related substances in dronedarone hydrochloride, and the newmethod was validated by the test.2. Residual Solvents: Headspace GC methodwas adopted to check the residual solvents of dronedarone hydrochloride，andthe method validation was done.3. Assay: The HPLC method was adopted todeterminate the content of dronedarone hydrochloride, and the new methodwas validated by the experiment.Results:1. Related substances:（1）The best chromatographic conditionwas chosen.（2）The theoretical plates of peak of dronedarone hydrochloridewere1240⁷. The resolution between dronedarone hydrochloride and the nearrelated substance was more than1.5.（3）After dronedarone hydrochloridewere treated by acid, alkali, oxidation, heat, light and high moisture, thedegradation products could be base-line separated from the main peak（.4）Thelowest detection limit was0.17ng.（5）The RSD of the main peak areas inreproducibility test was0.0⁶%. The RSD of the peak of biggest singleimpurity in repeatability test and in intermediate precision test were3.56%and3.69%, respectively. The RSD of peak of total impurities in repeatabilitytest and in intermediate precision test were2.0⁶%and3.68%, respectively（.6）Stability of the test solution: The RSD of the main peak areas, the peak ofbiggest single impurity and total impurities of were0.22%,3.38%and4.12%,respectively.（7）The RSD of the peak of biggest single impurity and totalimpurities in durability test were3.42%and4.05%,respectively.（8）Thebiggest single impurity in the three batches of samples were0.0826%,0.0860%and0.0865%, respectively. The total impurities of samples were0.2489%,0.2664%and0.2567%, respectively.2. Residual Solvents:（1）Thebest chromatographic separation condition was chosen.（2）The theoreticalplates of5kinds of residual solvents were more than50000. The resolutions between these solvents were more than2.5（.3）The lowest detection limit of5kinds of residual solvents were1.5μg·mL^-1,0.5μg·mL^-1,0.5μg·mL^-1,2.0μg·mL^-1and0.5μg·mL^-1, respectively. The lowest quantification limit of5kinds of residual solvents were4.0μg·mL^-1,3.0μg·mL^-1,3.0μg·mL^-1,4.5μg·mL^-1and1.0μg·mL^-1, respectively. The calibration curves wereY=1.198×10⁶X+1.811×10（3r=0.9997）, Y=1.862×10⁶X-4.135×10（3r=0.9996）,Y=2.118×10⁶X-7.605×10（3r=0.9996）, Y=1.555×10⁶X+7.244×10（2r=0.9999）and Y=5.845×10⁶X-4.154×10⁴, respectively.（5）In repeatability test, theRSD of the peaks areas of5kinds of residual solvents were1.82%,1.48%,1.97%,1.99%and1.93%, respectively. The RSD of the peaks areas of ethylacetate of the tested sample in repeatability test was4.47%. The RSD of thepeaks areas of ethyl acetate of the tested sample in intermediate precision testwas5.68%.（6）Their average recovery of five residual solvents were between97.57%and99.81%. The RSD of diversity were between1.41%and1.88%.（7）The RSD of the peaks areas of5kinds of residual solvents in the testedsamples were2.88%,3.46%,3.58%,3.79%and3.49%, respectively. The testsolution was stable in12hours.（8）The content of ethyl acetate in the testedsamples were0.0⁴83%,0.0⁴69%and0.0⁴85%, respectively.3. Assay（:1）Thebest chromatographic separation condition was chosen.（2）The theoreticalplates of main peak of dronedarone hydrochloride were24016. The tailingfactor of main peak was1.269.（3）The calibration curve for main peak wasY=1.902×10⁷X+3.165×10³, the correlation coefficient was1.0000.（4）Thelowest quantification limit of dronedarone hydrochloride was0.1ng.（5）TheRSD of reproducibility test, repeatability test and intermediate precision testwere0.11%,0.35%and0.61%, respectively.（6）In solution stability test, theRSD of the main peak area was0.09%. The test solution was stable in12hours（.7）Their average recovery rate for the assay was99.74%，and the RSDwas0.46%.（8）The RSD of durability test in the assay was0.55%.（9）Thecontents of three batches of samples of dronedarone hydrochloride were100.8%,100.7%and99.76%, respectively.Conclutions： The three methods of dronedarone hydrochloride for related substances detction, residual solvents inspection and assay wereestablished. By validation, the three new methods were proved to be specific,accurate and sensitive. These newly established methods may be used in thenew drug development process for quality control and stability study.Part two Studies on Drug Quality Control of film-coated tablet ofdronedarone hydrochlorideObjective: To study physical and chemical properties of film-coatedtablet of dronedarone hydrochloride which relate to its quality, to establishnew methods and offer datas for quality control.Methods:1. Related substances: The HPLC method was adopted todetect the related substances of film-coated tablet of dronedaronehydrochloride, and the new method was validated.2. Assay: The HPLCmethod was used to determinate the content of film-coated tablet ofdronedarone hydrochloride, and the new method was validated by theexperiment.3. Dissolution curve: To study dissolution characteristics ofself-made sample and original product in different dissolution media, thedissolution curve was established and the dissolution profiles evaluated by f₂similar factor.Results:1. Related substances:（1）The best chromatographic separationcondition was chosen.（2）The theoretical plates of peak of film-coated tabletof dronedarone hydrochloride were13679. The resolution betweendronedarone hydrochloride and near related substance was more than1.5.（3）After film-coated tablet of dronedarone hydrochloride were treated by acid,alkali, oxidation, heat, light and high moisture, the degradation products couldbe base-line separated from the main peak （.4）The lowest detection limit was0.17ng（.5）The RSD of main peak in reproducibility test was0.09%. The RSDof peak of the biggest single impurity in repeatability test and intermediateprecision test were3.83%and3.88%, respectively. The RSD of peak of totalimpurities in repeatability test and intermediate precision test were3.16%and3.84%, respectively.（6）Solution stability test：The RSD of the peak areas ofmain peak, the biggest single impurity and total impurities of were0.12%, 4.73%and3.63%, respectively.（7）The RSD of peak of the biggest singleimpurity and total impurities in durability test were4.68%and3.80%,respectively.（8）The biggest single impurity of three batches of samples were0.0144%,0.0151%and0.0153%, respectively. The total impurities of threebatches of samples were0.0⁴87%,0.0⁴82%and0.0⁴95%, respectively.2.Assay:（1）The best chromatographic separation condition was chosen.（2）The theoretical plates of peak of dronedarone hydrochloride were230⁶4. Thetailing factor of main peak of dronedarone hydrochloride was1.152.（3）Thecalibration curve for assay was Y=2.017×10⁷X+2.714×10⁴, the correlationcoefficient was0.9999.（4）The lowest quantification limit was0.1ng.（5）The RSD of main peak in reproducibility test, repeatability test andintermediate precision test were0.11%,1.17%and0.51%, respectively.（6）In solution stability test, the RSD of the peaks areas was0.16%.（7）Theaverage recovery rate was99.94%, and the RSD of the test was1.25%.（8）The RSD of durability test were0.37%.（9）The contents of three batches ofsamples of film-coated tablet were99.75%,99.72%and100.3%, respectively.3. Dissolution curve（:1）In hydrochloric acid solution（pH1.2）, The f₂similarfactors were61.42,59.96and60.36, respectively. All of f₂factors were morethan50. The study demonstrated that dissolution profiles of film-coated tabletof the self-made sample were similar to that of original product.（2）Inphosphate buffer（pH4.5）, the f₂similar factors were64.11,68.70and70.30,respectively. All of these f₂similar factors were more than50. The dissolutioncurves of the self-made sample were similar to that of original product.（3）Inphosphate buffer（pH6.8）, the f₂similar factors were70.84,71.70and71.72,respectively. Because of these f₂similar factors were more than50, thedissolution characteristics of self-made sample were similar to that of originalproduct.（4）In water, the f₂similar factors were69.48,73.16and71.75,respectively. All of f₂factors were more than50. The study demonstrated thatthe dissolution profiles of self-made sample in film-coated tablet were similarto that of original product.Conclusions：The methods to determine the related substance, and content of dronedarone hydrochloride film-coated tablet were established, andthe dissolution profiles of film-coated tablet of dronedarone hydrochloridewere studied. The results of this article study may be used in the new drugdevelopment process for quality control and stability study.Part three Determination of the impurities of dronedaronehydrochloride by liquid chromatography-massspectrometryObjective: To analyse the impurities of dronedarone hydrochloridequalitatively by liquid chromatography-mass spectrometry and deduce thestructure of impurity.Methods: A high performance liquid chromatography-tandem massspectrometric method was established for the analysis of impurities ofdronedarone hydrochloride. Mass spectrometric detection was operated on atriple quadrupole tandem mass spectrometer in positive mode. By optimizingthe parameters, the method to analyse the impurities of dronedaronehydrochloride qualitatively was established.Results: HPLC：The HPLC separation was performed by gradientelution on an Agela Venusil XBP-CN analytical column（4.6×250mm,5μm）.The flow rate was1.0mL·min^-1. The mobile phase A was10mmol·L^-1ammonium acetate（pH3.0）. The mobile phase B was acetonitrile. From0.01min to10min, the phase B changed from38%to44%. From10.01min to60min, the B was44%. The column temperature was set at35℃. The injectionvolume was20μL. MS：By using electrospray ionzation and positive ionmonitoring, the main impurities of dronedarone hydrochloride were analysed.The temperature of the desolvation and the ion source were set at350℃and115℃respectively. The flow rate of cone gas（N₂） was700L·Hr^-1. Theionization voltage and cone voltage were3.0kV and30V respectively.According to the mass spectra, it can be concluded that the impurity A was theoxide of dronedarone, and the impurity B was the debutylated compound ofdronedarone, and the impurity C was the starting material in the synthesisprocess, and the impurity D was the demethylated compound of dronedarone. Conclusions：This study established the method to analyse the impuritiesof dronedarone hydrochloride qualitatively by liquid chromatography-massspectrometry technology for the first time, which provides the evidences foranalysis of impurities. The method could be used in qulity control ofdronedarone hydrochloride.