Study On The Preparation Technology And Quality Standard Of Tongze Sanjie Granule

PurposeBenign prostatic hyperplasia(BPH)is one of a commonly clinical disease in middle-aged and elderly males,while its incidence is as high as 70 percent in men over 50 years old[1].Along with our country entering the aged society gradually,BPH is becoming a common factor that influences elderly males’ quality and lever of life[2].The main symptoms of BPH are two main symptoms,bladder irritation sign(frequent micturition,urgent urination,increased urine and urge incontinence)and obstructive symptoms produced by hyperplasia of prostate blocking urinary passage,have a non-negligible affect on patients’ quality of life.Tongze Sanjie is clinical experience prescription composed by Cinnamomum cassia Pres1,Paeonia lactilora Pall.,Curcuma kwangsiensis S.G.Lee et C.F.Liang,Prunus persica(L.)Batsch,Paeonia suffruticosa Andr.and other 5 constituents.It has curative effect on warming Yang and transforming Qi,promoting blood circulation and removing blood stasis and dissipating phlegm and resolving masses,and is used for the treatment of BPH(such as dysuresia,urine waiting,frequent night urination and retention of urine),which is caused of kidney Yang or Qi deficiency and phlegm and stasis syndrome.As the decoction,Tongze Sanjie has the shortcoming of inconvenience,easy moldy deterioration,lack of compliance and Hirudo nipponica Whitman in this prescription makes it fishy.Besides,the quality is difficult to control.Granule,a modern preparation,is convenient for taking,carrying and storage while retaining curative effect of decoction.And gradule is easier to take for patients after modifying the taste[3].So under the guidance of traditional Chinese medical theory,according to the requirement of new drugs registration and combining modern prepatation techniques,we will study the preparation and quality standards of Tongze Sanjie in this paper to meet the requirement of safety,effectiveness,uniformity and stability of new drugs.MethodsIn this paper,the preparation process and quality standard of Tongze Sanjie Granules were studied.The details are as follows:1.Preparation of Tongze Sanjie GranulesAt first,process route was determined by document research and pre-experiment.Process parameters of determined process route were optimized:The transfer rate of paeonol was selected as the evaluation index to study the amount of water and the amount of distillate in extraction of paeonol with the method of steam distillation.The transfer rate of Paeoniflorin was selected as the evaluation index to study the extraction times in the method of reflux extraction on dregs;the extraction of Paeonia lactilora Pall.and other 8 constituents:The transfer rate of Paeoniflorin was selected as the evaluation index to study the amounts of solvent,extraction time and extraction times by orthogonal test.The refining process,concentrating process and drying process conditions were determined by single factor investigation.With the paeonol encapsulation efficiency as the evaluation index,the inclusion process of paeonol with β-cyclodextrin(β-CD)was studied by the ratio of β-CD and paeonol,the ratio of β-CD and water and inclusion time as study factors and by ball grinding method and orthogonal test.And the process of making Tongze Sanjie granules was determined through the study of the molding process.2.Study on Quality Standard of Tongze Sanjie GranulesThe method of qualitative identification and quantitative determination of the compound Tongze Sanjie granules was established.Paeonia lactilora Pall.,Paeonia suffruticosa Andr.,Prunus persica(L.)Batsch,Lycopus lucidus Turcz.var.hirtus Regel and Curcuma kwangsiensis S.G.Lee et C.F.Liang were identified by thin-layer chromatography(TLC).The content determination conditions of the index components of paeoniflorin and paeonal were established by high performance liquid chromatography.Establish inspection items of Tongze Sanjie Granules according to the general of Chinese pharmacopoeia(volume IV).Results1.Preparation of Tongze Sanjie GranulesPre-experiment of process routes showed results were as follows:oil-water mixtures extracted in route 1(Cinnamomum cassia Pres1,Curcuma kwangsiensis S.G.Lee et C.F.Liang and Paeonia suffruticosa Andr.was extracted naphtha and then their dregs were extracted by water,other 7 constituents were extracted by water)were difficult to separate,which made industrial production complex;So we chose route 2(Paeonia suffruticosa Andr.extracted by steam distillation and then their dregs were extracted by water,other 9 constituents were extracted by water).We finally determine the preparation process of Tongze Sanjie granules by process parameters optimization and molding process:The Paeonia suffruticosa Andr.was extracted by steam distillation with 12 times of water and collecting 8 times of distillate,after 24-hour cold storage,crystallization,suction filtration(mother liquid was reserved),desiccation in desiccator,paeonol was obtained and then was produced to inclusion complex.Aqueous solution afer steam distillation centrifuged(2500r/min,30min)while hot,and centrifugate was reserved.Dregs of Paeonia suffruticosa Andr.boiled with 7 times of water,extract centrifuged(2500r/min,30min)while hot,and centrifugate was merged with centrifugate above and reserved.Other 9 herbs boiled with water twice,extract centrifuged(2500r/min,30min)while hot,and centrifugate concentrated.Centrifugate merging with centrifugate of Paeonia suffruticosa Andr.and mother liquid was concentrated until density was about 1.15(65℃).Thick paste was desiccated by vacuum drying oven(80℃,0.09MPa).Combined dry extract above with β-cyclodextrin inclusion compound of paeonol,crushed over 80 mesh sieve,then added dextrin and 90%ethanol as a wetting agent to make granule.After drying,the Tongze Sanjie Granules were observed.2.Study on Quality Standard of Tongze Sanjie GranulesPaeonia lactilora Pall.,Paeonia suffruticosa Andr.,Prunus persica(L.)Batsch,Lycopus lucidus Turcz.var.hirtus Regel and Curcuma kwangsiensis S.G.Lee et C.F.Liang were identified by thin-ayer chromatography(TLC).The conditions of the determination of the paeoniflorin and paeonal was that acetonitrile for the mobile phase A,the 0.1%phosphoric acid for the mobile phase B,and for gradient elution.The elution conditions are as follows:0～15 min 84%→78%B,15～25 min 78%→55%B,25～30 min 55%→40%B.The detection wavelength of paeoniflorin was 230 nm,and the detection wavelength of paeonal was 274 nm.The column temperature was 30℃.The injection volume was 10 μL.Flow rate 1.0 mL·min-1.Check exterior,granularity,water content,dissolubility.ConclusionIn this study,the extraction and molding process of Tongze Sanjie Granules were studied,and a stable and feasible granules preparation process was established.Check the project of granules to meet the relevant requirements and qualitative identification of Tongze Sanjie Granules and quantitative determination of the content of the component,which can be controlled the quality of finished products.