Cai Huang, Preparation And Anti-inflammatory Effects Of The Freeze-dried Powder

Chaihuang lyophilized powder for injection was first prepared.Chaihuang lyophilized powder for injection was prepared from Chaihu volatile oil which was extract from dried root of Bupleurum Chinese DC(Umbelliferae) by water vapor distillation and baicalin which was extract from Scutellaria baicalensis Georgi.Based on quality standard of Chaihu injection and Shuanghuanglian lyophilized powder for injection,the formulation of Chaihuang lyopholized powder for injection was established,the formulation consists of Chaihu volatile oil 3 g,baicalin 20 g, Hydroxypropyl-beta-cyclodextrin 100 g,glucose 75 g,and water for injection to 800 ml,then pH was adjusted to 5.4-6.4 with 10%NaOH to make baicalin dissolved and total volume is 1000 mL,subpackaged to 500 vials(0.5 mg n-hexanoic acid per vial) and lyophilized.Tentative usage and dosage was 1 vial/time,once to twice daily intravenously.Each vial contains baicalin(C₂₁H₁₈O₁₁)30-36 mg and n-hexanoic 0.45-0.55 mg.A gas chromatography method for content determination of volatile oil in Bupleurum Chinese DC and n-hexanoic acid in Chaihuang lyophilized powder for injection was established,this method has high sensitivity and no excipient interferences,n-hexanoic acid was linear over the range of 0.02-1.2 mg/ml,the RSDs of precision,reproducibility and recovery were all less than 5%(n=5).Volatile oils from Bupleurum Chinese DC of different places were analyzed by GC-MS,20 peaks were isolated and more than 10 compounds were determined,in which 1,1,4,7-tetramethyl-1a,2,3,4,4a, 5,6,7b-octahydro-1H-cyclopropa[e]azulene was first found in Bupleurum Chinese DC. The content of n-hexanoic acid,one of the active constituents in volatile oil of Bupleurum Chinese DC,corresponded with the results that determined by the content-determining method we had established. The method of content determination of baicalin by HPLC was developed,this method had high sensitivity,baicalin was linear over the range of 6-48μg/g,the RSDs of precision,reproducibility and recovery were all less than 5%(n=5).In the preparation of Chaihuang lyophilized powder for injection,extract process was optimized according to the content of n-hexanoic acid,and orthogonal test was also developed.The optimum process for extract of volatile oil from Bupleurum Chinese DC was proportion of material to solvent 1:6,soak time 24 h and extract fraction 600 ml. The optimum process for extract of baicalin was proportion of material to solvent 1:10, extraction time 90 min and 3 time extraction.Product inspection and stability of Chaihuang lyophilized powder for injection were also established according the content of n-hexanoic acid and baicalin,the results showed the product inspection and stability of lyophilized powder for injection we had prepared according to our process met all the criteria.Good anti-inflammatory effect of Chaihuang lyophilized powder for injection and n-hexanoic acid was demonstrated by xylene-induced ear edema in mice.Optimum effective time of hydrocortisone and optimum time of xylene-induced ear edema were also verified,xylene painting method was also improved.