| Background:Traditional Chinese medicine as an important treasure of Chinese traditional medicine,occupies an important place in drug therapy,but with the vigorous development of the cause of Chinese medicine,Chinese medicine formula particles gradually took to the stage of new era,single Chinese herbal pieces by water extraction,separation,concentration,drying,granulating of grain,under the guidance of TCM theory,according to the clinical prescription of traditional Chinese medicine after deployment,for use in patients with a blunt.Compared with traditional single Chinese medicine decoction pieces,Chinese medicine formula granules have the advantages of safety,effectiveness,economy,simplicity and portability,and are widely used in clinical practice.Verbena,as one of the clinical commonly used traditional Chinese medicine,under the guidance of the theory of TCM,widely applied in the treatment of various diseases,and play an important role in current,the lack of provincial and national level verbena formula granules quality standards,is not conducive to clinical widely used,for the quality control and promote the in-depth research.Objective:We selected the conform to the requirements of the"Chinese pharmacopoeia"2020 version of decoction pieces from verbena,first of all to control verbena formula granule preparation technology and optimization,and through the cooperation with enterprises of verbena pilot and mass production parameters optimized formula particles,secondly,to conduct of verbena formula particles identification,examination,determination of extract of;Finally,choice hastatoside and verbenalin as verbena formula an important index of grain quality standard research,through the research on feature maps and content determination,specificity and good system applicability,high accuracy and durability good verbena formula granules quality standard research methods,for the development and utilization of verbena and quality standards.Methods:1.Study on preparation technology of Verbena formula granules:According to the standard decoction of Verbena,the extract rate,index component content and transfer rate,and the consistency of characteristic spectrum were selected as the indexes;choose（the batch number:P2103001,Xiang yang,Hubei province）as the research sample,from the water addition（compared with the number of Chinese medicinal slices;10,11,12 times）,decoction times（1,2,3times）,decoction time（1,1.5,2 hours）three factors,respectively take three levels of each factor,through orthogonal test,determine the extraction,solid-liquid separation,concentration,drying,granulating process,through the three batch of pilot validation,The mass transfer and material balance in the preparation of verbena intermediates and finished products were investigated,and the preparation technology of verbena formula granules was finally determined.2.Identification of verbena formula granules:According to the method of verbena[Identification]in Chinese Pharmacopoeia 2020 edition,the control medicinal materials of Verbena and the standard products of hastatoside and verbenalin were selected as controls.By preparing test solution,reference solution,developing agent and color developing agent,the influence factors of verbena formula particles,such as sample quantity,specificity,different thin layer plate and developing temperature,were investigated.3.Examination and study of verbena formula granules:According to the method of verbena[Inspection]in The Chinese Pharmacopoeia 2020 edition,the particle size,moisture,solubility and volume difference of the formula particles of Verbena were examined.4.Examination and study on extract of Verbena formula granules:Each group was determined by hot immersion method under the determination method of alcohol-soluble extract（General Rule 2201 of Chinese Pharmacopoeia 2020 Edition）.5.Study on fingerprint/characteristic Atlas of Verbena formula particles:（1）Firstly,on the basis of the research results of verbena standard decoction,several different chromatographic conditions were investigated according to the chemical composition of Verbena and the results of reference research,and better liquid phase conditions were obtained to get as many characteristic peaks as possible in as short analysis time as possible.Secondly,DAD detector was used to scan at 200-400nm to determine the best wavelength.Finally,different chromatographic columns were used to detect the same sample,so as to obtain a column with high efficiency.In summary,the optimal chromatographic conditions were determined.（2）Choose verbena formula particles as the research sample,from the extract method,ultrasound,reflux),extraction solvent（water,methanol,ethanol）,extraction time（15,30,45,60 min）,extraction times（1,2,3times）four factors respectively,each factor in different levels,through the test to determine the variety of inspection plan,choose the best treatment method;（3）According to the optimized UPLC chromatographic conditions,the blank（excipient）group and each experimental group were blank control,Yin and Yang control,to investigate specificity;At the same time,the precision,stability,repeatability and durability of the sample solution were compared by using the same sample for multiple injection detection,detection at different times and detection in multiple groups.In summary,this study established fingerprint method of verbena formula granules from multiple perspectives and carried out methodological verification.6.Determination method of Verbena formula granules:By optimizing HPLC methodological conditions and preparation conditions of sample solution,the contents of hastatoside and verbenalin in3 batches of Verbena formula granules were determined,and the methodological verification was carried out.Results:1.Preparation process of verbena formula granules:the influencing factors were as follows:Factor B（extraction times）>factor A（extraction time）>factor C（multiple of water addition）.The content of group A2B2C2was the highest,that is,adding 11 times of water（relative to the prepared slices of Verbena）.2.TLC identification:The study showed that verbena formula granules had good specificity,and the TLC conditions（point sample size of 8μl,efficient thin layer plate）were suitable for developing at different temperatures（4℃,25℃）.3.Granule examination:The experimental results show that all the indexes are in conformity with the requirements.4.Extractum:The results showed that the RSD of extract content of Verbena was small and in line with the regulation.5.Pre-treatment method of the test product solution:Take an appropriate amount of the product,fine,take about 0.1g,weigh it accurately,put it into a tapered bottle with a plug,add 100ml water,ultrasound（power 250W,frequency 40k Hz）for 30min,and treat once,then it can be prepared.Study on fingerprint of Verbena granule formula:The method was proved to be stable,reliable and reproducibility,which provided a basis for the establishment of the quality standard of Verbena granule.6.Verbena granule content:Three batches of verbena granules consist of hastatoside（the content ratio are 2.88%,2.82%and 2.90%）and verbenalin（the content ratio are 1.77%,1.64%and 1.59%）.Conclusion:The quality standard of verbena formula granules was established successfully,which provides important reference for the quality evaluation of verbena formula granules. |
PDF Full Text Request