Studies On Chemical System And New Dosage Form Of Compound Holly Bark Preparation

ObjectThe aim of this study was to offer technical support for further exploitation of Compound Holly Bark preparation,explore the chemical component of the crude drug in Compound Holly Bark Capsule,expect to find some monomer component,improve quality control of the main medicinal materials and their preparations by chemical and quality research.Then an assessment of qualities standard system can be normalized and the level of quality control can be improved during the production.Meanwhile,new dosage forms were expected to research to adapt to different population especially children.Methods and ResultsSeverinia buxifolia(Poir.)Ten.,is mainly distributed in South China.It is belonging to the Rutaceae family,and is widely distributed in the Guangdong and Guangxi Province. The dried root and stem of is used as a drug in mainland.However,there are seldom report about the constituents of it,which would restrict the development of it and its preparation. In order to find out bioactive constituents,the chloroform -soluble fraction and petroleum ether -soluble of ethanol extract of Severinia buxifolia(Poir.)Ten.Was subjected to chromatographic columns filled respectively with silica gel,reserved-phase(RP)-silica gel(C18),Sephadex LH-20 gel,and/or separated by preparative HPLC.Six compounds were obtained.On the basis of physical and chemical properties as well as detail spectroscopic analysis(UV,1HNMR,13CNMR,El-MS,DEPT,HMQC,HMBC),these compounds have been identified as compoundⅠBuxifoliadine-B,compoundⅡand compoundⅢare non-named,compoundⅣ- Behenic Acid,compoundⅤβ-Sitosterol and compoundⅥDaucosterol.Among these,compoundⅡand compoundⅢwere obtained from this Chinese herb for the first time. Current quality of Severinia buxifolia(Poir.)Ten.and its preparation were still determined by a method of TLC identification and using reference medicinal materials as a marker substance.The level of the quality is so low that it cannot control the crude drug and its preparation better.Therefore,in order to analyze the constituents of Severinia buxifolia(Poir.)Ten.Root and stem,buxifoliadine-B,a principal constituent of the heral drug,was assigned as a marker compound.It can be detected by silica gel TLC with petroleum ether(60～90℃)-ethyl acetate -abs.EtOH(6:3.5:0.5)as developing solvent system.The TLC spots developed were clear troubleless and easy to recognize.The liguid has a good effect as a developer.Chromatography is stable in the property.The reversed phase HPLC,which carried out through a C- 18 column Diamonsil (250mm×4.6mm,5μm),with a mobile phase methanol-water(89:11),at a flow rate 1.0 ml/min and detective wavelength 264nm for assay of buxifoliadine-B,showed good linearity in the range of 0.05～0.75μg with a correlation coefficient 0.9999.The average recovery was 100.7%,RSD was 1.79%.it was found that the content of buxifoliadine-B in the samples were 0.1043 mg·g-1,0.0968 mg·g-1,0.0923 mg·g-1.Qualitative and quantitative analysis method for the bioactive constituent Buxifoliadine-B in the roots and stems of Severinia buxifolia(Poir.)Ten.Was built up for the first time,the results also provided a basis for the qualitative control of the Severinia buxifolia(Poir.)Ten.root and stem.Cortex Ilicis Rotundae has a high medical value and was used in clinic as many kinds of dosage forms widely.But Current quality of Cortex Ilicis Rotundae and its preparation were still determined by a method of TLC identification and using reference medicinal materials as a marker substance.The quality standard must be improved to control the crude drug and its preparation better.Therefore,in order to analyze the constituents of Cortex Ilicis Rotundae bark,Syringin,a principal constituent of the herbal drug,was assigned as a marker compound.The reversed phase HPLC,which carried out through a C-18 column Diamonsil(250mm×4.6mm,5μm),with a mobile phase acetonitrile-water -water(15:85),at a flow rate 1.0 ml/min and detective wavelength 264nm for assay of Syringin,showed a good linearity in the range of 3.06～15.30μg with a correlation coefficient 0.9991.The average recovery was 100.3%,RSD was 2.876%.it was found that the everage content of Syringin in the samples was 20.2431mg/g,RSD is 2.297%.Then the samples from different producing area were 7.8366 mg/mg·g-1,20.9669 mg·g-1,20.3286 mg·g-1,25.7848 mg·g-1.Qualitative and quantitative analysis method for the bioactive constituent Syringin in the bark of Cortex Ilicis Rotundae.was built up for the first time, the results also provided a basis for the qualitative control of the Cortex Ilicis Rotundae. Bark.But the content limit of Syringin should not be evaluated because of the different producing area and the lost of Syringin during production.As for the Cortex Ilicis Rotundae. Bark the content cannot be below 0.7%of Syringin free of water. Syringin was used to build a determination method of the Compound Holly Bark Capsule basis on the method of controlling the Cortex Ilicis Rotundae.Bark,which chromatographic conditions was used too.Syringin in the Compound Holly Bark Capsule showed a good linearity in the range of 0.256～4.608μg with a correlation coefficient 0.9999.The average recovery was 100.71%,RSD was 1.66%.According to the data often batches of sample from production,the content limit was determined that the Cortex Ilicis Rotundae.Bark the content cannot be below 0.25mg Syringin free of water per milliliter sample.Then as for Compound Holly Bark Capsule the content cannot be below 1.80mg Syringin free of water per capsule.Technique of making Compound Holly Bark Effervescent Granules extract powder, including extraction,separation,drying,was uniformed to the original standard of Compound Holly Bark Capsule.But the water in the extract powder must be controlled below 3%.Proper excipient was chose by single factor analysis.So Polyethylene glycol 6000(PEG 6000),a good soluble polymer with non-toxic side-effects,neutral,unique physical and chemical properties,and good solubility,was used at first to separate and protect the acid/alkali part from the anther one free water,which can take neutralization reaction if put together with water.The results showed that citric acid is in liquid which cannot be inclusion by PEG6000,but the inclusion sodium bicarbonate was difficult to be grinded and sieved after dried.Soft wood suitability,excipient dosage,drying time,the granules characters were chose as index system.Other granulation methods was explored that different concentration of PEG 6000-anhydrous ethanol solution used as an adhesive,but the 0.5%%one showed the best quality.Different honey were tried to use as adhesive but none can be good to effervescent granules,different concentration of PVP-K30 with anhydrous ethanol were suitable for granulation,but PVP-K30 is more expentsive,easily in moistrure absorption than PEG6000.so PEG6000 was a good choice as excipient.Different ration of sodium bicarbonate to citric acid were chose in process of granulating as effervescent granules disintegrant ratio.The index of effervescent time,PH value and taste were observed to evaluate the better disintegrant ratio,which was 1:1.3.The filler choice showed that different filler took almost the same action to the soluble and different to the clarity.Dextrin was the best one but most cheap among the fillers lactose and mannitol.The orthogonal test design was adopted to study the pharmaceutical technology of Compound Holly Bark Effervescent Granules.Excipient ratio which affected the effervescent time by using orthogonal design method.The result showed that PEG6000 and dextrin were the good adhesive and filler.So the shortest effervescent time happened in the condition were PEG6000,dextrin and the better disintegrant ratio 1:1.3 about sodium bicarbonate and citric acid.Three kinds of raw material in Compound Holly Bark Effervescent Granules,Holly Bark,Cyperus Tuber,were identified by TLC according the original standard of capsules. The result showed the method of TLC Identification was feasible.Syringin was used to build a determination HPLC method of Compound Holly Bark Effervescent Granules basis on the method of controlling the Cortex Ilicis Rotundae.Bark and Compound Holly Bark Capsule,which chromatographic conditions was used too,but anhydrous ethanol was used as the pretreatment solvent.Syringin in the Compound Holly Bark Effervescent Granules showed a good linearity in the range of 0.10～4.00μg with a correlation coefficient 0.9999. The average recovery was 102.1%,RSD was 0.6%.ConclusionsSix compounds were got from the crude drug of Severinia buxifolia(Poir.)Ten.. CompoundⅡand compoundⅢwere new,found and obtained from this Chinese herb for the first time.CompoundⅠbuxifoliadine-B can be used as marker in HPLC method for determination of the content of crude drug,which is convenient for controlling the quality during production and assessment the quality from different material area.HPLC method,for determination of the content of syringin in Holly bark and its preparation was built and could be used stable,strong specificity and good reproducibility. It is possible for building and regulating assessment system of Holly bark and its preparation,which could be used to control and chose different crude drug from different area.The research of compound Holly bark effervescent granules gave a preliminary pharmaceutical technology and quality standard,a good guide for further pilot magnification.Then the research of new dosage form was very suitable for children and might solve the problem of simple product.Series product of the system will be manufactured because of meeting the market demand.In a word,this study gave an important basis for deep development of compound Holly bark and controlling the quality of the product from the chemical,quality and new dosage forms.