| The ancient classic"Duliang Fang" consists of two traditional Chinese medicines,Baizhi and Chuanxiong,and has a definite curative effect on tension headache.The volatile oil component is the main effective part of Duliangfang,but it is unstable,poor in water solubility,and is not easy to be made into oral solid preparation.In this thesis,the chemical composition and extraction and purification methods of the volatile oil components of Duliang Pills were systematically studied.On this basis,two oral preparations of volatile oil microcapsules(capsules)and volatile oil cyclodextrin inclusions(capsules)were prepared and stabilized.The two formulations were compared and analyzed in three aspects:sex,in vitro release and in vivo pharmacokinetics.The optimal dosage form was Duliang Pill volatile oil microcapsules(capsules).The research results of this thesis lay the foundation for the research and development of Duliangfang new drug,which is of great significance for improving the clinical efficacy of Duliangfang.It can also provide ideas and methodological references for other traditional Chinese medicine pre:parations containing volatile oil.1.Literature ResearchThrough reviewing relevant research literatures at home and abroad in recent years,this paper summarizes the tension headache and Duliang,and studies the current situation and existing problems of the chemical constituents and pharmacological effects of the volatile oils in the prescriptions,and the inclusion technology and microcapsule technology of Chinese medicine volatile oil.The review provides a literature basis for the experimental research of the paper.2.Analysis of chemical constituents of volatile oil from DuliangfangIn this part,GC-MS was used to compare and analyze the volatile oil components of Radix Paeoniae Alba,Rhizoma Chuanxiong and Duliangfang,and the index components which can be used for process screening and quality control were selected,and the HPLC method was used to determine the volatile oil in Duliangfang.Ester content.A total of 24 components were identified from the volatile oil extracted from Rhizoma Chuanxiong.The main chemical components were ligustilide(44.73%),Yangchuan ligustilide(16.745%),butyl azlactone(8.9688%),m-toluene acetate.(3.1115%),eucalyptus(1.8398%),decyl ene alcohol(1.3889%),etc.,accounted for 81.571%of the total volatile oil.A total of 17 components were identified from the volatile oil extracted from the white peony root.The main chemical components were dodecanol(27.123%),myristyl alcohol(22.864%),palmitic acid(21.14%),and 1-tetradecene(2.6301).%),myristic acid(2.5414%),etc.,accounting for 86.615%of the total volatile oil.A total of 49 components were identified from the volatile oil of Duliangfang.The main chemical components were ligustilide(38.293%),butyl azlactone(8.6047%),1-decene(7.0919%),and decylene(3.4438).%),4-terpineol(2.6021%),decyl ene alcohol(2.0169%),palmitic acid(1.7943%),etc.,accounting for 81.577%of the total volatile oil.By comparing the chemical constituents of single and compound volatile oils,the results showed that the main components of the compound volatile oil were derived from the volatile oil of Rhizoma Chuanxiong,and the content of ligustilide was the highest.White peony is a multi-base original medicine.In this part,GC-MS was used to qualitatively compare and analyze the white carp of different bases and different producing areas,but the chemical composition was different,and the suitable quantitative index components were lacking.The content of ligustilide in Duliangfang volatile oil was the highest.The pharmacological action was obvious.Therefore,the main choice of ligustilide as the main parameter component of the process parameters and quality control of Duliangfang volatile oil was established.The HPLC method was applied to determine the content of ligustilide in the volatile oil of Duliangfang The chromatographic conditions were finalized as:ZORBAX SB-C18(250 mm × 4.6 mm,5 ?m);mobile phase:methanol-water(60:40,V/V);flow rate:0.8 mL/min;detection wavelength:330 Nm;injection volume:10.L;column temperature:30 ?The linear range of ligustilide was determined to be 0.003313?0.053mg/mL(R=0.9994,n=5),and the recovery rate was 99.4%(RSD=1.53%).The method has high sensitivity and accuracy and good repeatability.It can be used for quality control of Duliangfang volatile oil3.Duliangfang volatile oil extraction process researchTaking the single-flavored Chuanxiong medicinal materials as the research object,the traits,oil extraction rate and ligustilide content were used as the evaluation indexes,and the steam distillation method(B method),supercritical C02 extraction method and salting-out auxiliary steam distillation method were compared.The enzymatic hydrolysis-assisted steam distillation method was applied to the extraction of volatile oil from Ligusticum chuanxiong Hort.The results showed that the best extraction method was salting-out assisted steam distillation.The single factor test method combined with orthogonal test was used to evaluate the volatile oil extraction rate and ligustilide content as the evaluation index.The comminuted degree of the medicinal material,the amount of water added,the soaking time,the extraction time and the amount of sodium chloride were studied.The impact of the extraction.Finally,the best extraction process for the volatile oil of Duliangfang is as follows:crushing 24 mesh sieve,soaking for 4 hours,adding 12 times sodium chloride aqueous solution,sodium chloride dosage is 8%of the quality of the medicine,and extracting for 8 hours,both Liangfangfang volatile oil The rate was 0.22%(mL/g)and the average content of ligustilide was 61.0%4.Study on the preparation technology and quality control of Duliangfang volatile oil ?-CD inclusion compound capsuleFirstly,a method for the determination of volatile oil in the volatile oil inclusion complex of Duliangfang by ultraviolet spectrophotometry was established.The main evaluation indexes were the yield of inclusion compound,the utilization rate of volatile oil and the oil content of inclusion compound.Orthogonal experiments were carried out to compare the inclusion materials of ?-cyclodextrin and hydroxypropyl-?-cyclodextrin,as well as the inclusion effects of four preparation methods:ultrasonic method,grinding method,magnetic stirring method and high-speed dispersion method.Finally,the optimal inclusion material is?-cyclodextrin.The best preparation method is high-speed dispersion method.The optimal preparation parameters are:the ratio of volatile oil to ?-cyclodextrin is 1:10(g:g),package The combined temperature was 50 0 C and the inclusion time was 5 min.The results showed that the yield of the inclusion compound was 72.7%,the inclusion rate was 63.5%,and the oil content was 8.57%.The yield of the inclusion compound obtained by laboratory test was 86.72%,and the inclusion rate was 91.39%The dosage is 9.58%,which further proves that the inclusion process is stable and feasible;The quality of P-CD inclusion complex of Duliangfang volatile oil was evaluated by microscopy,infrared spectroscopy(IR),differential scanning calorimetry(DSC),thin layer identification,gas chromatography,etc.Things have indeed formed.The composition analysis of the volatile oil in the inclusion compound by GC-MS analysis showed that the inclusion technique had little effect on the composition of the main chemical components of the volatile oil,but the composition content changed.Taking Duliangfang volatile oil inclusion compound as an intermediate,taking moisture absorption,moldability and fluidity as indicators,the molding process parameters of pharmaceutical excipients,wetting agent concentration and drug auxiliary ratio were investigated,and the optimal molding was finally selected.The process parameters are as follows:lactose is an auxiliary material with a ratio of 1:2 and a wetting agent concentration of 70%ethanol.The granules are prepared and placed in a capsule shell,0.3 g per capsuleA method for the determination of ligustilide in the preparation of "Duliangfang Volatile Oil Inclusion Capsule,and the inclusion complex intermediate was established by HPLC method.Based on this,the quality standard of "Duliangfang Volatile Oil Inclusion Capsule" was established5.Study on the preparation technology and quality of Duliangfang volatile oil microcapsules(capsules)Firstly,a method for the determination of volatile oil in the microcapsules of Duliangfang volatile oil was established by ultraviolet spectrophotometry.The microcapsule yield,drug loading and embedding rate were used to investigate the microcapsules.Preparation of microcapsules by factors such as wall material type,wall material ratio,core wall material ratio,liquid material concentration,additive dosage,emulsification time,emulsification speed,inlet air temperature,feed power,atomization pressure,fan power,etc.The final prescription for the determination ofthe volatile oil microcapsules of Duliangfang is acacia-maltodextrin(2:1),core-capsule(1:8),the concentration of the liquid is 15%,and the dosage of the additive PEG6000 The optimum emulsification process is emulsification time 9 min,emulsification speed is 10 rpm/min;the optimal spray drying process is inlet air temperature 185 ° C,feed power 12%,atomization pressure 35 MPa,fan power is 100%.The average yield of volatile oil microcapsules of Duliangfang was 74.7%,the embedding rate was 72.2%,and the drug loading was 9.67%.The microcapsule yield obtained by laboratory test was 68.89%,and the embedding rate was 52.09%.The dosage was 8.42%,which further verified the stability and feasibility of the process.The microcapsules were evaluated by electron microscopy,particle size determination,thin layer identification and gas chromatography.The results showed that the microcapsules prepared under the optimal process conditions were round,dense and uniform in particle size distribution.It has a good normal distribution,and the particle size distribution ranges from 1.446 ?m to 19.510 ?m,and the average particle diameter is 9.532?m.The composition of the prepared microcapsules was analyzed by GC-MS.The results showed that the microencapsulation of volatile oil by spray drying method had little effect on the main chemical composition of volatile oil,but the content of the components changed.Taking Duliangfang volatile oil microcapsule as an intermediate,taking moisture absorption,moldability and fluidity as indicators,the molding process parameters of pharmaceutical excipients,wetting agent concentration and drug auxiliary ratio were investigated,and the optimal molding process was finally selected.The parameters are:lactose as an auxiliary material,the ratio of the drug is 1:2,the concentration of the wetting agent is 80%ethanol,and the granules are prepared and placed in a capsule shell,0.3 g per capsule.A method for the determination of ligustilide in the preparation of "Duliangfang volatile oil microcapsules(capsules)"and microcapsule intermediates was established by HPLC method.Based on this,the quality standard of "Duliangfang volatile oil microcapsules(capsules)" was established.draft.6.Stability,in vitro release and pharmacokinetics of two formulations of Duliangfang volatile oilThe stability of Duliangfang volatile oil and its preparations were studied by influencing factors test,accelerated test and long-term test.The results showed that(1)The total content of volatile oils decreased under high temperature,high humidity and strong light.The high temperature condition has the greatest influence on the volatile oil,and the total content of the main components is reduced from 91.8%to 67.5%.The main active ingredient,ligustilide,is reduced from 48.6%to 5.73%,which is reduced by about 88.2%.Under high humidity and strong light irradiation conditions.The content of ligustilide decreased by about half,while that of butylphthalide,butenyl lactone and decyl ene alcohol increased.(2)The content of these components and other main components under the conditions of high temperature,high humidity and strong light irradiation after the inclusion of volatile oil as clathrate and microcapsules is not significant,indicating that the volatile oil is made into inclusion complex and microcapsules.Its stability has been significantly improved.(3)Accelerated test and long-term test The preliminary research results further show that the stability of the volatile oil is significantly improved after it is made into inclusion complex and microcapsuleThe results of in vitro release of Duliangfang volatile oil and its preparations showed that after the volatile oil was made into inclusion complex and microcapsules,its release in artificial gastric juice and artificial intestinal juice could be improved,and it was found that all of Duliangfang volatile oil was made.After microcapsules,the in vitro release rate was significantly higher than that of the inclusion complex in artificial gastric juice and artificial intestinal fluid.The cumulative release of Duliangfang volatile oil microcapsules in artificial gastric juice for 90 min reached 76.43%,which was 53.15 times that of Duliangfang volatile oil(1.438%)at the same time,which was 5.239 of Duliangfang volatile oil inclusion compound(14.59%)Times.In addition,in the artificial intestinal fluid,the cumulative release of the volatile oil microcapsules of Duliangfang was 39.93%within 90 min,which was 28.32 times of the volatile oil of Duliangfang(1.41%)in the same time,which was 5.523 times of the volatile oil inclusion complex of Duliangfang(7.23%);the cumulative release of Duliangfang volatile oil inclusion compound in artificial gastric juice within 90 min is 14.59%,which is 10.15 times of Duliangfang volatile oil(1.438%);in artificial intestinal juice,Duliangfang volatile oil inclusion The cumulative release of the substance within 90 min was 7.23%,which was 5.128 times that of Duliangfang volatile oil(1.41%).The pharmacokinetics study of the volatile oil and its preparation in Duliangfang showed that:(1)comparison peak time(Tmax),inclusion complex group(0.94±0.632)h,microcapsule group(1.653±2.245)h peak time is longer than that of the volatile oil group(0.694±0.356)h,indicating that it takes a certain time for the volatile oil to be encapsulated and microcapsules to be released in rats.After the rats are filled with the inclusion complex and microcapsules The absorption is slower than the absorption of volatile oil.(2)Contrast peak concentration(Cmax),peak concentration of microcapsule group(8.686±2.924)?g/mL higher than volatile oil group(6.113±3.927)?g/mL,and peak concentration of inclusion compound group(0.653±0.453)?g/mL is much lower than the volatile oil group and the microcapsule group,indicating that the microcapsule can promote the absorption of volatile oil,while the inclusion compound reduces the absorption of volatile oil;(3)the elimination half-life(T1/2z),relative to the volatile oil group(5.06±3.392)h,the microcapsule group(20.302±43.595)h stayed in the rats for a prolonged time,eliminating the slowing,while the inclusion compound group(2.923±1.320)h eliminated faster,indicating that the volatile oil was made into microcapsules.After that,it can prolong the action time of the drug,and it can play a sustained release effect,and can better exert the drug effect.However,the individuals in the microcapsule group had large differences.(4)Area under the curve of the comparison drug time AUC(0-?),microcapsule group(58.666±34.679)h*mg/L>volatile oil group(53.738±53.677)h*mg/L>inclusion compound group(1.396±0.661)h*mg/L,the results showed that compared with volatile oil,the bioavailability of microcapsules increased,while the bioavailability of inclusion complex decreased,indicating that the bioavailability of microcapsules was higher for Duliangfang volatile oil.Inclusion compound.(5)Comparing the relative bioavailability,the results showed that the volatile oil was encapsulated and microencapsulated into a solid to improve the oral bioavailability of the volatile oil of Duliangfang,but the bioavailability of the inclusion complex was not obvious,while the bioavailability of the microcapsule was significant.Increasing,the relative bioavailability of microcapsules(67.002%)is higher than that of inclusion complexes(2.762%),which is 24.26 times that of clathrates,indicating that the use of Duliangfang volatile oil as microcapsules can significantly improve its bioavailability.The effect of themicrocapsules is better than that of the inclusion complex.The results of in vitro release studies have similarities with the results of in vivo pharmacokinetic studies,indicating that Duliangfang volatile oil is not suitable for powdering by inclusion technology,and microencapsulation may be the best method.According to the above research results,the optimal dosage form is finally selected as a microcapsule.The microcapsule can be used as a final dosage form or as a preparation intermediate,and further processed into tablets,capsules,injections,ophthalmic preparations,patches,aerosols and suspensions,etc.;The indication is a tension headache.Therefore,the microcapsule can be further made into a patch,an aerosol,etc.,so that it can be better applied to the clinic. |
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