Preparation Of Reference Materials Of Dihydrotanshinone Ⅰ And Miltirone

Salvia miltiorrhiza Bunge is one of the most widely used traditional Chinese herbal medicines.It has the function of promoting blood circulation and relieving qi deficiency,and can be used to produce preparations for treating cardiovascular and cerebrovascular diseases.In addition to the traditional functions of promoting blood circulation,calming nerves,relieving pain,nourishing the heart and calming the nerves,tanshinone has been increasingly researched in recent years on its anti-oxidation,anti-bacterial,anti-inflammatory,cardiovascular and cerebrovascular diseases,anti-tumor and other pharmacological effects.Has a very broad application prospects.At present,there are many scientific reports on the separation and preparation of the fat-soluble component tanshinones in Salvia miltiorrhiza Bunge.However,the separation and preparation methods lead to different quality and purity of tanshinone.This study optimizes the salvian ketones which are important in the process of Danshen and related industrialization.The standard sample preparation process of the substances(danshen neosaponin and dihydrotanshinone I),with Danshen neosinone and dihydrotanshinone I as the target,developed a national-level certified standard sample with traceable and quantitative transfer function,and established effective Fixed value analysis method.The main research contents are as follows:1.Preparation of standard samples of dihydrotanshinone I and miltironeThe roots of S.miltiorrhiza was extracted and purified by column chromatography.The standard samples of dihydrotanshinone I and miltirone were used as reference for the standard samples of dihydrotanshinone I and miltirone.The optimized preparation process is as follows:reflux extraction times 3 times,ethanol volume:5 to 7 times,ethanol concentration 95%,crystallization temperature:2 to 5℃,salvia miltiorrhiza eluate:petroleum ether-benzene,dihydrotanshinone I wash Deliquoring:petroleum ether-dichloromethane.2.Determination of purity and structural confirmation of standard samples of dihydrotanshinone I and miltironeThe standard sample of dihydrotanshinone I and salviane ketide obtained above was determined by HPLC-DAD,and the purity was over 99%.Four major spectra(NMR NMR.LC-MS LC/MS,IR spectroscopy,UV UV spectroscopy)were used to identify the structure,and the samples obtained were confirmed to be miltirone and dihydrotanshinone I.3.Uniformity and stability test of standard samples of dihydrotanshinone I and miltirone The prepared standard samples of Danshen neosinone and dihydrotanshinone I were stored in a volume of 250 μg/mL with a chromatographic grade of acetonitrile as a solvent for uniformity and stability test The peak area of high performance liquid chroma to graphy was used.The purity was determined by normalization method,and the results were statistically calculated by one-way ANOVA(F test)and trend analysis(T test).The results showed that the preparation of miltirone and Dihydrotanshinone I standard samples was stable and evenly distributed.Stable,good solution stability and long-term stability.The uncertainty of the uniformity test of dihydrotanshinone I standard sample was 0.013128,and the extended uncertainty was 0.104.The uniformity test uncertainty of Danshen neoketone standard sample was 0.0235,and the extended uncertainty was 0.1344.4.Determination of standard samples of dihydrotanshinone I and miltironeAccording to the requirements of GB/T15000.3-2008,the standard samples of dihydrotanshinone I and Danshenxin were sent to test 8 qualified laboratory laboratories for the fixed value experiment.After statistical calculation,the standard values and uncertainties of dihydrotanshinone I and miltirone were obtained.The final result is:the standard value of the standard sample of Danshenxin is 99.35%,the uncertainty is 0.3347%,the standard value of the standard sample of dihydrotanshinone I is 99.71%,and the uncertainty is 0.26%.The final result is:the standard value of the standard sample of miltirone is 99.35%,the uncertainty is 0.3347%,the standard value of the standard sample of dihydrotanshinone I is 99.71%,and the uncertainty is 0.26%.