Quality Standards For Cyclovirobuxine D And Huangyangning Tablets

Cyclovirobuxine D(CVB-D)is the therapeutic alkaloids extracted from the plant of Buxusmicrophylla Sieb.et Zucc.var.sinica Rehd.et Wils.and its congeners using acidic ethanol then alkaline hexane and is finally gotten by recrystalization.CVB-D is also the active pharmaceutical ingredient(API)of Huangyangning tablets which is a good clinical medicine in the treatment of coronary heart disease and angina.In view of the current quality standards of Huangyangning Tablets and its API,especially the controversial issues and the outstanding focus of the industry in terms of standard revision.Based on the improvement of drug safety,the quality standard of Huangyangning tablets and its API and strengthening the quality control,several research was carried out in the following aspects.(1)The research of quality standard for cyclovirobuxine D.The solubility of the listed active pharmaceutical ingredient(API)of Huangyangning tablets was studied.As a result,the active pharmaceutical ingredient(API)of Huangyangning tablets is freely soluble in chloroform,soluble in methanol,sparingly soluble in ethanol,slightly soluble in acetone,and insoluble or almost insoluble in water.The research modified the project of identification,changing the physical and chemical reaction to liquid chromatography.Quantitative and qualitative analyses and limitation of the impurities.This method can separated and detected 12 homologen peaks except CVB-D peak(11).Among them,there are 2 major impurities for the higher content,cyclobuxine D and cyclobuxine D double bond hydrogenation The compond of the 4 impulities in the remaining 10 tiny impurities were speculated and the molecular formula of the 5 impulities in the remaining 10 tiny impurities were speculated and the 7 tiny impulities in the remaining 10 tiny impurities were quantitated due to the main content was more than 0.1%.so finally 9 impurities including the 2 major impurities got qualitative.The research modified the project of examination,using liquid chromatography and calculating with area normalized percentage method.According to the date,the amount of impurities in the API would be a 15.5%,two major impurity content limits set at 10.0%and 3.0%respectively.The research added the detection of residual organic solvent.5 kinds of solvents residues were detected according to Pharmacopoeia,and the method was verified.Establishment of determination method of the main component in the API of Huangyangning tablets.The suitable agent was selected for pre-column derivatization to derivatize main component and all homologen impurities in the API.RP-HPLC method was adopted to analyze the derivatives,C8 column was used for separation and acetonitrile-0.01%formic acid solution(71:29)as mobile phase,the detection wavelength was at 230 nm.The flow rate was 1.0 mL/min and the column temperature was 25℃.Results The new method can separate cyclovirobuxine D from impurities with resolutions of more than 1.5 and up to 10 homologen impurity peaks were detected and 9 were further quantitatively and qualitatively analyzed among the ten,LOD and LOQ of cyclovirobuxine D were 0.35 ng(RSD=2.22%,n=5)and 1.36 ng(RSD=4.55%,n=5)respectively,and the average recovery was 102.52%(RSD=1.40%,n=6).As a result,the content of CVB-D in the 10 batches of samples from four manufacturers was 78.28%～85.48%,and the average value was 82.78%.The content of the impurities were 10.16%～15.79%,and the average value was 13.54%.Using four kinds of HPLC methods to determine the main component content in the active pharmaceutical ingredient of Huangyangning tablets,and carry on the comparison and evaluation.As a result,limitation required for assay of the main compound is no less than 75.0%in the active pharmaceutical ingredient of Huangyangning tablets.The comparison of manufacturing technique and the chemical and physical properties between the API and the unlisted API of high purity.Due to the API is the key to the quality of the product and the controversy over the standard,and to evaluate whether the unlisted API of high purity can replace the API of low purity,a comparation has been made between the API of low purity in the actual feeding and the unlisted API of high purity in appearance,solubility,main component content,impurity content,fingerprint of impurities,tablet dissolution,near infrared detection of tablet.(2)The reasearch of quality standard for Huangyangning tablets.The uniformity determination of tablets can use the liquid chromatography.Calculation of content uniformity determination of tablets can use the calculation of A+2.2S in general information in Phamacopoeia 2015 to replace each theory calculation in the current pharmacopoeia standards,which should be 20.0.Establishment of determination method of the main component in Huangyangning tablets.The method is consistent with the method of determining the main component of the API,and the methodological validation results are good.The content of CVB-D in the 57 batches of samples from five manufacturers was 55.22%-82.86%,and the average value was 69.10%.Labeled amount limitation required for assay of the main compound is specifications(1)no less than 0.375 mg/tablet,(2)no less than 0.75mg/tablet.The Limitation required for assay of the main compound of Huangyangning tablet should be 90.0%to 110.0%of the labeled amount.