The Study On The Preparation Technology And Quality Standard Of Zhenkuang Jieyu Capsule

Research background: Zhenkuang Jieyu Capsule herbs was based Diankuang Mengxing Decoction,which was the classical representative famous prescription established by the famous medical expert Wang Qingren of the Qing Dynasty.It was used for many years as experienced prescription,in which were Peach Kernel,Red Peony Root and Rhizoma cyperi,ete.It had a obvious curative effect. In addition,it possessed activating blood circulation to dissipate blood stasis and eliminating phlegm and resuscitation efficacy,it was used to treat manicdepressive syndrome that caused by Phlegm Stagnatio with stagnation of blood stasis.To provide a beneficial reference for the study of the traditional Chinese preparation used to treat mental disorder diseases,we developed it into capsule in order to use easily in clinical application.Objective:To study the preparation technology and quality standard of Zhenkuang Jieyu Capsule.The extraction yield of solid extracts and major active ingredient content were chosen as indexes, the orthogonal test was used to optimize the extraction technology and determine the molding technology of preparation in the preparation technology aspect.To study the main drugs identification and assay methods of active constituent in the prescription,to establish the effective and controlled quality standard in quality aspect.Methods: 1 The study on the preparation technology of Zhenkuang Jieyu CapsuleThe decoction times,decoction time and water amount were chosen as the major influence of the extraction technology, The extraction yield of solid extracts and major active ingredient content were chosen as indexes, the orthogonal test was used to screen the optimal extraction process and condition by comprehensive assessment;The study on granulation process and the bulk density,hygroscopicity, mobility and capsule routine examination item,etc. as indexes were used to determine the molding technology of preparation. 1.1 The study on the extraction technology 1.1.1 The study on the water extraction technologyThe extraction yield of solid extracts and the content of Paeoniflorin were chosen as indexes,the orthogonal test according to L9(34)table was used to screen the optimal extraction process and condition by comprehensive assessment,under the condition of 3 levels from each of factors, which are the water amount(A),decoction time(B) and decoction times(C). 1.1.2 The study on separation and purification technologyIn order to reduce oral dosage and guarantee efficacy in drug therapy,the water extract mixed other impurities such as starch, phlegmatic temperament,etc. was purified because of the large dosage prescription dose.Considering production conditions of pharmaceutical industries,alcohol deposit was chosen to remove impurities by the orthogonal test.The extraction yield of alcohol solid extracts and the content of Paeoniflorin were chosen as indexes,the orthogonal test was used to screen the optimal process by comprehensive the two indexes,under the condition of 3 levels from each of factors, which are the relative density of extraction solution(A) and alcohol content(B),in order to optimize the alcohol density with the lower extraction yield of solid extracts and higher water content,at last the repeat the validation test was does using the selected optimum technical condition. 1.2 Study on molding technologyGranulating process test was used to study granular formability;The granular fluidity was studied by determining angle of repose;The packing volumes was studied by filled into 0# capsule with different mesh granule and drug powder;The hygroscopic was investigated by comparing the size of humidity absorption between granular and drug powder; The molding technology was entrenched by taking into account granular formability, fluidity,packing volumes,daily dose and hygroscopic. 2 Study on quality standardThe main drugs in the prescription was identified by TLC, the content of active constituent in the prescription was determinate by HPLC.Project standard was made accordingly in accordance with capsules recorded in the appendix of China Pharmacopeia( 2010 edition, Volume I). 2.1 IdentificationTLC method was adopted to qualitatively identify Peach Kernel,Red Peony Root,Rhizoma cyperi and Radix Bupleuri,etc.monarch drug in the prescription in accordance with appendices VIB recorded in the appendix of China Pharmacopeia( 2010 edition, Volume I) and referencing to other literatures. 2.2 Content determination 2.2.1 The choice of chromatogram conditionsThe choice of the determination wavelength:using appropriate paeoniflorin as the control, and was scanned under spanning wavelengths from 200 to 400 nanometers in accordance with appendices A Ⅳ recorded in the appendix of China Pharmacopeia( 2010 edition, Volume II).The choice of the mobile phase:octadecyl silane for the bonded silica filler,organic modifier including the amount of methanol, acetonitrile and p H regulators glacial acetic acid was studied based on literatures. 2.2.2 The choice of extract methodThe result of the ultrasonic extraction using methanol, ethanol and water as the solvents indicates the content in water extract is higher,but water extract is more difficult to filter, with too many impurity peak,so the method which remove the impurities by extraction of n-butanol after water extract was choiced,the optimal extraction method was established by study on water amount,ultrasonic time and extraction times. 2.2.3 Study of the methodologyStudy of the methodology was conducted by linear ranges,repeated test,precision test,stability test of solution and recovery rate test,etc.Results: 1 The study on the preparation technology of Zhenkuang Jieyu Capsule 1.1 The study on the extraction technology 1.1.1 The study on the water extraction technologyThe order of the various factors on the extraction technology was B>C>A(shown in Table 2).Considering the actual productions situation,the optimal combination of three factors was defined as A2B2C2. 1.1.2 The study on separation and purification technologyTaking extractive yield after ethanol precipitationas index,choiced A3B3 as the optimal conditions for separating and purifying.choiced the content of Paeoniflorin as indexes,the optimal combination was defined as A2B3. 1.2 Study on molding technologyThe molding technology was established as using automatic capsule filling machine filled 60 mesh granule into 0#capsule,average packing volumes was 0.5g,daily dose was 12 capsules taking three times by taking into account granular fluidity, hygroscopic,packing volumes and dose. 2 Study on quality standard 2.1 IdentificationThe results of TLC identification indicated that four identified herbs(Peach Kernel,Red Peony Root,Rhizoma cyperi and Radix Bupleuri)could show the same color pot as the control herbs or contrast in the same position,the negative controlled substance did not show interference. 2.2 Content determination 2.2.1 The choice of chromatogram conditionsThe choice of the determination wavelength:Paeoniflorin have absorption peaks in 230 nanometers,so the wavelength of determination was 230.The choice of the mobile phase:using the mobile phase of methanol-acetonitrile-5 mmol/L KH2PO4(15:12:80:0.1)as mobile phase,Keep the Paeoniflorin peak in this product was separated completely from the other peaks to meet the requirement of determining. 2.2.2 The choice of extract methodTaking the contents of the product under the gross deviation, blending, Weighing accurately 0.5 g, and set in the conical flask, adding accurately 30 ml distilled water into it,infiltrating for 20 minute, ultrasonicing for 25 minute, transfering into separatory funnel, using n-butanol extraction 3 times, each time 30 ml, combinating of n-butanol extract, volatilizing to dryness in boiling water, with methanol dissolving and exact transferring to 50 ml volumetric flask, adding methanol to scale, shaking well, with the micro pore filtering film(0.45 μm) filtratiing, the subsequent filtrate can be obtained 2.2.3 Study of the methodologylinear ranges:The relationship between the Paeoniflorin peak area(Y) and the concentrations of Paeoniflorin was evaluated by linear regression,and derive linear equation:Y=30634.4582C+1644.0135, r=0.9999,it indicated Paeoniflorin had a good linearity relationship within the concentration range from 207844 μg/ml to 44.5500 μg/ml.the test solution was stable within 12 hour,repeatability and precision were good,recovery met the requirement.Conclusion:The preparation technology determined by the experiment is stable,reasonable,and can provide reliable basis for production.The quality standard is simple,effective and suitable for the quality control.