Studies On The Production Process And Quality Standards For Total Terpenoids Of Atractylodes Lances

AimsThrough comparative studies on the quality of the medicinal material of AtractylodesLances,to select its best sources or bases,then to achieve the following objectives:(1)Toscreen the effective parts of the medicinal material of Atractylodes Lances;(2)To optimizethe production process of the effective parts of the medicinal material of AtractylodesLances;(3)To analyze the chemical composition of the effective parts of the medicinalmaterial of Atractylodes Lances;(4) To study the quality standards and the stability of theTotal Terpenoids of Atractylodes LancesMethodsQuality study of the medicinal material of Atractylodes Lances:According to standardsof the “Chinese Pharmacopoeia” and relative references,we studied the quality ofAtractylodes Lances medicinal material in the items of characteristic identification,physicaland chemical identification,total ash,moisture and assay in order to find out the bestresource.Choice of the effective parts of Atractylodes Lance:Three parts were obtained throughdifferent extracting and separating of the medicinal materials of Atractylodes Lances,thentheir pharmacodynamic screening were tested and the extract of the total Terpeoids ofAtractylodes Lance through steam distillation was confirmed as the effective part.Optimization of production process of the effective parts of AtractylodesLance:Using steam distillation as extraction method,basing on production costs,conditionsof production and extraction methods,we selected the water volume for extraction,soaktime,extraction time and size as factors and optimized the production process by orthogonal experimental design method.Then total terpenoids production rate was used as theevaluation index to test the the optimum extraction conditions.Study of chemical constituents of the effective parts of Atractylodes Lance:GC-MSmethod was carried in this chemical constituent study.The chromatographic conditions wereas follows:BPX5(25m×0.22mm×0.25μm)as the analytical column;temperatureprogramming;the starting temperature of60°C,keep the temperature3min,heated to150°Cat the speed of10°C/min,then3°C/min rate of heating to220°C,and then heated to250°C at25°C/min,maintaining this temperature for2min;the carrier gas flow rate was1ml/min;split ratio was15:1.MS conditions were as follows:EI source;the electron energy70ev;ion source temperature of200°C;interface temperature230°C;detector voltage400v;the mass range10~550amu,and Xcalibur Chemstation NIST standard massspectrometry Gallery retrieval.Study of quality standards and the stability of the total Terpenoids of AtractylodesLance:The items of quality study included characteristic identification, physical andchemical identification and GC identification,moisture,burning residual,heavy metal andarsenic salt check and determination. The chromatographic quantitative method wasestablished according to the guidelines of “Chinese Pharmacopoeia”(2005edt. and2010edt.).Taking hinesol and β-eudesmol as references;1-Pentadecanol as the internal standard,the total Terpenoids of Atractylodes Lance was determined.At the same time, theexperiments of stability were carried out through investigating the factors such aslight,heat,humidity, the accelerated test(40°C±2℃, relative humidity75%±5%)andsample reserved test.ResultsThe quality study of the Atractylodes Lances medicinal material showed: the contentsof hinesol and β–eudesmol in medicinal material of Atractylodes Lancesl from differenthabitats varies greatly,of which the total maximum value of hinesol and β–eudesmolreached6.029percent,and the minimum value was0.032percent;the content of hinesolranged between0.007~3.673percent,and β–eudesmol0.022~2.356percent. Among thosemedicinal materials, the herb habitated in Hubei and Shanxi province had higher contents of hinesol and β–eudesmol and lower content of total ash.Compared with Shanxiprovince,Hubei Yingshan County is richer in natural resources with huge production andstable supply.In order to keep quality stability and supply continuity of the originalmedicinal.The herb natived in Hubei province was selected as the medicinal material forextraction of total terpenoids.Screening of the effective parts of Atractylodes Lances: Pharmacological efficacyexperiments showed the extract of total terpenoids by steam distillation could reduce theamount of gastric juice,lower the free acid,total acidity and five-hour emissions of totalacid,so we chose steam distillation as the extraction method.The effective part was theextract of total terpenoids of Atractylodes Lances by steam distillation method.Studies of production process of the effective parts of Atractylode Lances:Accordingto the production costs,production conditions and extraction methods,we chose the watervolume for extraction(A),soak time(B),extraction time(C)and size(D)of four factors andoptimized the production process by orthogonal design.Taking production rate of totalterpenoids as evaluation index,the best extraction process was confirmed for A3B2C2D3,thatwas size(6mesh sieve),the appropriate amount,8times the amount of water,immersion for3hours and steam distillation extraction for8hours.According to the analysis ofvariance,extraction time and the amount of water had significant effects on the yield of totalterpenoids;the immersion time and particle size had no significant impact,and each level ofvariance was extremely small.Considering production costs and the time cycle,we changedthe best process into A3B3C2D3,that was,size(6mesh sieve),appropriate amount,8times theamount of water,steam distillation for8hours.Studies of chemical constituents of the effective parts of Atractylodes Lances showedthat: Twenty components were identified from the total terpenoids of AtractylodesLances.The content of hinesol,β-eudesmol, γ-eudesmol,α-eudesmol and2,3-Dihydro-1-oxo-phenalene were higher than1%,of which hinesol and β-eudesmol were87%.In addition,therelative content of sesquiterpenoids were1.79%.Studies of quality standards and the stability of the total Terpenoids of AtractylodesLance showed that: The total Terpenoids of Atractylodes Lance was oily solid with pale yellow to brown,smell good specificity, bitter and xin and freely soluble in ethyl acetate andn-hexane.The items of identification all showed the positive reaction,and it had lowercontent in the items of moisure,residue on ignition,heavy metals and arsenic salt.For thedeterminationt,we used hinesol and β-eudesmol as an index component, the GC as themethod,and carried on the methodology examination with accurate,convenient andeasily.Its precision, reproducibility and recovery all conformed to the guidlines oftraditional Chinese medicine quality standards of the“Chinese Pharmacopoeia”(2010edt.)XVIII A.Studies on stability of total terpenoids of Atractylodes Lances included:the influencefactors test,the accelerated test and sample reserved test.The influence factors test showedthe total terpenoids at light,high humidity and high temperature conditions had nosignificant changes in the content,namely,that it was relatively stable in theseconditions.Accelerated test results showed that the total terpenoids which be placed for sixmonths at the temperature of40°C±2°C, relative humidity of75%±5%were conformedin all indicators.Sample reserved test results showed total terpenoids could be placed atroom temperature for at lease18months.Conclusion①In that paper, the best source of medicinal material was confirmed,and the identificationmethods for Atractylodes Lances medicinal material were developed.②The effectiveparts of Atractylodes Lances were found out,at the same time, through theexperiment,best extraction process was optimized.③GC-MS method was carried outand twenty components were identified from the effective parts of the medicinalmaterial of Atractylodes Lances.The areas normalization method showed the identifiedcompounds consisted of more than99percent of the total terpenoids.④Studied on thequality of total terpenoids of Atractylodes Lances and developed a draft quality standardof total terpenoids of Atractylodes Lances.Established the gas chromatography methodsfor the first time for hinesol and β-eudesmol for the fifth sort of new drugs.Themethodology inspection results indicated that the method,accurate and sensitive,withgood linear relationship, high precision and recovery,could be used as the quality control for total terpenoids of Atractylodes Lances.