Study On Preparation And Quality Analysis Of Compound Xuejie Gel

Objective:To develop a gel preparation for Compound Xuejie Gel.This preparation not only has a nice stability and a good adhesion to skin,but also has higher contents.The preparation would be studied systematicly on its extraction technology,manufacturing technology,quality standards,in vitro percutaneous absorption, formulation variables and Fingerprints.Methods:1 The research of the extraction technology:The SFE-CO₂ extraction process for Rhizoma Corydalis was optimized by L₉(3⁴) orthogonal design experiment,in which the yield of tetrahydropalmatine was regarded as the index observed and three factors including extraction pressure,temperature and extracted time were considered and the content of tetrahydropalmatine was determined by HPLC.The SFE-CO₂ extraction process for Angelica was optimized by L₉(3⁴) orthogonal design experiment,in which the yield of volatile oil and ferulic acid from were regarded as the markers and the content of ferulic acid was determined by HPLC and the four influence factors such as extraction pressure,temperature, extraction time and the size of angelica powder were considered and the L₉(3⁴) table was used.The amount of volatile oil extracted from Olibanum by distillation method was regarded as the index of chemical compositions extracted to optimize the extraction process of Olibanum by orthogonal design experiment in which three influence factors such as amount of water,soaking time and extraction time were considered and the L₉(3⁴) table was used.2 The study on the manufacturing technology:Base on the results of the literature consulting and Pre-experiment,the kinds and the use range of matrixes were determined for Compound Xuejie Gel.The manufacturing technology for the gel was optimized by L₉(3⁴) orthogonal design by determinating the content of xuejie and scoring synthetically on some other aspects such as the adhesive ability,spreading ability,glossiness and uniformity together as indexes.3 The study on the quality standards for the preparation:The Sanguis Draconis,Angelica,Rhizoma Corydalis,Olibanum and Borneolum Synthetieum in the gel preparation were identified by TLC.The contents of tetrahydropalmatine and ferulic acid in the gel preparation were determined by HPLC respectively.The chromatography conditions of tetrahydropalmatine were that the Agilent ZORBAX SB-C₁₈(5μm,4.6 mm×250 mm) column was used;that mobile phase was composed of methanol-0.1% phosphoric acid solution(adjust the pH to 6.0 with triethylamine)(60:40); the flow rate was 0.8mL·min^-1 with UV detection at 280nm and that the column temperature was 35℃.The chromatography conditions of were that the Agilent ZORBAX SB-C₁₈(5μm,4.6 mm×250 mm) column was used;that mobile phase was composed of methanol-0.3%aqueous acetic acid(35:65);the flow rate was 1.0 mL·min^-1 with UV detection at 320nm and that the column temperature was 35℃.The other subjects of the preparation were tested according to the method described for gel in 2005 derision China pharmacopeia below the relative volumns of appendix I Q. 4 The study of the in vitro percutaneous absorption:Franz permeating pull was used to carry out the relative experiments and the content of tetrahydropalmatine of percutaneous absorbed from Compound Xuejie Gel was determined by HPLC.5 The study on the impact of factors:Reference to "traditional Chinese medicine,natural medicine,the stability of the guiding principles of research techniques",the impact of factors was studied with the investigation items such as appearance,characteristics, identification,adhesiveability,uniformity,spreading ability and the content of tetrahydropalmatine.6 The study on the fingerprint for the preparation:Based on the fingerprint standard of quality control for compound Xuejie Gel by HPLC, the fingerprint picture was established,which includes the amount of common peaks,similarity and ratio of peak areas.Ten batches of samples were detected.Results:1 The optimum SFE-CO₂ extraction technology for Rhizoma Corydalis was that the ethanol of 95%was the entrainer and that the medicinal material powder,which was in size between 20 and 60 mu,was soaked by ethanol with amonia for 24 hours and then under the pressure of 25 Mpa at 55℃,was extracted for 1.5 hours.The transfer rate of tetrahydropalmatine was 92.14%and was more than that of ethanol extraction process and its rate of extract only was 0.665%.The optimum SFE-CO₂ extraction technology for angelica was that the ethanol of 75%was the entrainer and that under the pressure of 30 Mpa at 60℃,the medicinal material powder,which was in size between 20 and 60 mu,was was extracted for 2 hours.The optimum distillation extraction technology of volatile oil for Olibanumwas the medicinal material was soaked with water for 1 hour and then extracted with 10 times of water for 6 hours continuously.3 the quality standards:The TLC identification methods for each medical material in the gel preparation were simple,and the spots on TLC pictures were clear.Study on determination had shown that the linear rang of the concentration of tetrahydropalmatine was 0.1148μg～1.148μg(r=0.9999);that the precision test indicated that RSD was equal to 1.57%;that the repetition test indicated that the average content of tetrahydropalmatine in the patch was 0.1774 mg·g^-1 and the RSD was 1.79%;that the average recovery was 101.36%and the RSD was 2.01%and that he contents of tetrahydropalmatine in 3 batch of gels were 0.1782 mg·g^-1,0.1774 mg·g^-1 and 0.1737 mg·g^-1 respectively.Study on determination had shown that the linear rang of the concentration of ferulic acid was 0.031μg～0.310μg(r=0.9999);that the precision test indicated that RSD was equal to 1.57%;that the repetition test indicated that the average content of ferulic acid in the gel was 0.085 mg·g^-1 and the RSD was 1.71%;that the average recovery was 95.82%and the RSD was 2.01%and that the content of ferulic acid in 3 batch of gel were 0.0814mg·g^-1,0.0782 mg·g^-1 and 0.0795 mg·g^-1respectively. The adhesive ability and the pH of the gel preparation was accord with the regulation.4 The in vitro percutaneous absorption test:the cumulative in vitro permeation(mouse) rate within 12 hours was 78.48%.5 the results of impact of factors Experiment had shown that the temperature had significant impact to the compound xuejie gel and humidity and light exposure had no significant effect to the preparation. 6 Fingerprints picture of the compound xuejie gel:The preliminary HPLC fingerprint picture was set up for compound xuejie Gel,in which a total of 24 peaks were marked.Conclusions:The Compound Xuejie Gel possesses good stability,satisfactory adhesive property to skin and high content.Many research have been done in many different aspects,such as extraction technology, manufacturing technology,quality standards of the preparation, the cumulative permeation rate,the impact of factors and Fingerprints of compound.All of these reseaches lay a good foundation for the further development of new traditional Chinese medicine Compound Xuejie Gel.