Studies On Preparation Process For Guishao Capsules

The prescription of Guishao capsules came from JinkuiYaolue, which was written by Zhangzhongjing, a famous doctorin Ming-dynasty. It contains six kinds of herbal medicine suchas Radix Paeoniae Alba, Angelica sinensis, Rhizoma LigusticiChuanxiong, Rhizoma Atractylodis Macrocephalae, Poria andRhizoma Alismatis. It was recorded in traditional medicine thatit could be used to relieve hyperactivity of liver, nourish bloodand spleen, and disperse the pathogenic damp. It has shown inmany reports that Danggui Shaoyao powder has an effect ofdilating peripheral veins, especially brain veins since 1980 sothat it can improve external symptoms significantly. DangguiShaoyao powder is effective on brain metabolism, and it hasshowed favorable action in relieving Alzheimer's disease.Powder was recorded originally in Jinkui Yaolue, which hadmany disadvantages such as big dosage, slow absorption andlow absorption rate, etc. According to the technologicalrequirements about studying on new Chinese TraditionalMedicine, on the base of analyzing bioactive components andpharmacological effect of each traditional medicine, combiningwith the clinical requirements, category, active ingredients oreffective parts and its physical and chemical proprieties, thetraditional medicine was extracted so that chemical constituentswere isolated selectively in order to prepare appropriateformulation by the characteristics of semi-manufactured sample.Objective: To study the extracting process of herbalmedicine in order to optimize the best conditions; To explore theprocess of preparation for manufacturing Guishao capsules.Methods: Conditions of extraction were chosen by meansof orthogonal test according to the properties of the herbalmedicine itself and its physical and chemical proprieties to makecapsules. With the yield of volatile oil as indexes, water volume,soaking time and extracting time as factors, the optimalextracting condition for volatile oil from Radix AngelicaeSinensi, Rhizoma Chuanxiog and Rhizoma AtractylodisMacrocephalae was investigated. With the transferring rate ofpaeoniflorin as indexe, and the volume of water, extracting timeand times as factors, the optimal extracting process for theextracting with water was explored. Then every kind oftraditional medicine was extracted and parameter of preparationmoulding was studied to make capsules.Results: The optimal extraction condition of the volatile oil:Angelica sinensis, Rhizoma Ligustici Chuanxiong, RhizomaAtractylodis Macrocephalae were crushed into powderappropriately, 8 volumes of water were added and soaked for 3hours, and then extracted for 8 hours. The result for waterextraction: Radix Paeoniae Alba, Rhizoma Alismatis and Poriawere crushed into powder. The powder was combined withresidue after volatile oil was extracted and decocted with 12volumes of water for 1.5 hours for three times. The technologyof preparation of the capsules: The extracts were taken and dried,and then grinded into powder. Amylum was added to driedpowder and mixed. Ethanol was added to make into softmaterial and granules were made by using 18 mesh, and then thegranules were dried at 60~65℃. After the volatile oil wasdissolved in appropriate ethanol and was sprayed the powder,the granules were mixed and closed for half an hour. At last thegranules were filled into capsules.Conclusion: The optimal extraction and preparationtechnique for Guishao capsules were ascertained by usingorthogonal test.Safety, efficiency and quality controlled are elementaryproperties for medicine. Safety is premise, efficiency isobjective and quality control is guarantee. Quality control fordrug is one of its basic attributes and the basement on securityand validity of medicine at the same time. Quality standard is avital measure to monitor drug quality and also embodiment forquality control. In order to control internal quality of Guishaocapsules, its identification and determination were studied toestablish quality standard complying with the technologicalrequirements about studying on new Chinese TraditionalMedicine.Objective:To establish a series of TLC methods toidentify traditional Chinese medicine in Guishao capsules suchas Radix Paeoniae Alba, Angelica sinensis, Rhizoma LigusticiChuanxiong, Rhizoma Atractylodis Macrocephalae, RhizomaAlismatis and Poria. To explore a reversed-phase HPLCmethod for determination of albiflorin and paeoniflorin inGuishao capsules in order to provide one guideline for theirquality control.Methods: (1)Studies on identification methods:Ingredientsin Guishao capsules were identified with thin-layerchromatography. (2)Studies on determination method: ①Albiflorin and paeoniflorin from Guishao capsules wereextracted with methanol through supersonic wave, then theextract solution was filtered. ②Chromatogram condition:Choose appropriate chromatogram column and adjust theformulation, proportion and flow rate to separate the peaks ofalbiflorin and paeoniflorin well. ③Preparation of standardcurve: Prepare a series of the reference solution, and thendetermine peak area under above-mentioned condition. Theregression equation was obtained with the content of albiflorinand paeoniflorin as abscissa and the relevant peak area asordinate. ④Confirmation of the lowest detection limitation:Dilute the reference solution until the value of S/N was morethan 3. Under this circumstance, the relevant concentration wasthe lowest detection limitation. ⑤In the experiment ofreproducibility, prepare the sample solution and repeat fivetimes in the same way, respectively. Assay the concentrations ofthe solutions. ⑥In the experiment of precision, take the sametest solution, inject it into the apparatus for five times, determinethe peak areas of albiflorin and paeoniflorin, then calculate theconcentrations of albiflorin and paeoniflorin in comply with thesame standard curve with the same apparatus in the same day,record the chromatograms, and then calculate the precision. ⑦In the experiment of stability, transfer the sample solution fromGuishao capsule to determine the relevant peak area at 0, 2, 4, 6,8, 10, 12 and 24 hour, respectively. ⑧In the experiment ofrecovery, transfer 5 shares of Guishao capsules and add thereference solution, respectively. Detect the content of albiflorinand paeoniflorin of every sample. ⑨Determination: TransferGuishao capsules to prepare the test solutions and determine thecontent of albiflorin and paeoniflorin, respectively.Results: (1) Results of identification methods: Fourmethods were established for the identification of RadixPaeoniae Alba, Angelica sinensis, Rhizoma LigusticiChuanxiong, Rhizoma Atractylodis Macrocephalae, RhizomaAlismatis with TLC. The principal spot in the chromatogramobtained with the test sample was similar in position, colour andsize to the principal spot in the chromatogram obtained with thereference solution. Specific spots were not found for Poria bymeans of different TLC condition. There were not specific spotin the test solution compared with the reference solution andblank solution. So above the identification methods for one kindof herbal medicine was not recorded in the standard. (2) Resultof determination method: ①The method of supersonicwave-extraction was simple, quick and stable. ②Under theconditions of this experiment, the mobile phase wasacetonitrile–0.05% acetic acid (17:83) at the flow rate of 1.0ml·min-1, the detection wavelength was set at 230nm, and thevolume of injection was 20μl. Under the circumstance, thepeaks of these samples for albiflorin were separated well withthe resolution of no less than 3, the column efficiency is no lessthan 5000 theoretical plates, the tailing factors for the peak were1.21. The peaks of these samples for paeoniflorin wereseparated well with the resolution of no less than 2, the columnefficiency is no less than 4000 theoretical plates, the tailingfactors for the peak were 1.5. ③With the content of albiflorinand paeoniflorin as abscissa, the relevant peak area as ordinate,we got regression equation of albiflorin: Y=1×10~7X+10245,r=0.9999(n=5).The linear range is between 0.06072～0.1417mg?ml-1. The regression equation of paeoniflorin isY=2×10~7X-214638,r=0.9999(n=5). The linear range of isbetween 0.07968~0.2390mg ? ml-1. ④The lowest detectionlimitations of albiflorin and paeoniflorin were 31.80ng and20.40ng, respectively. ⑤In the experiment of reproducibility,determine albiflorin and paeoniflorin from Guishao capsulesfive times and we got the values of RSD of albiflorin andpaeoniflorin were both 0.49%. ⑥In the experiment ofprecision, take the solution of albiflorin and paeoniflorin fromGuishao capsules, repeat to make each concentration solutionfive times in the same way, inject each sample into the apparatusfive times, determine the peak areas of albiflorin andpaeoniflorin and calculate the concentrations of albiflorin andpaeoniflorin in comply with the same standard curve with thesame apparatus in the same day, record the chromatograms, thencalculate the within-day precision. The RSD of albiflorin andpaeoniflorin were 0.93% and 0.53%, respectively. ⑦In theexperiment of stability, transfer the reference solution of...