ABSTRACT
OBJECTIVE To prepare hyperoside mixed nanomicelles （Hyp-F127/TPGS） and optimize its preparation technology，and to investigate its intestinal absorption characteristics. METHODS Hyp-F127/TPGS was prepared by thin film dispersion method. Based on single factor test and Plackett-Burman design ，combined with Box-Behnken response surface method ， the preparation process was optimized and validated using entrapped efficiency （EE）and drug loading （DL）as evaluation indexes ， F127-TPGS mass ratio ，hydration time and the amount of Hyp as factors. The appearance and microscopic morphology of Hyp-F127/TPGS obtained by the optimal technology were observed ，and the particle size ，polydispersity index （PDI）and Zeta potential were also determined. The critical micelle concentration （CMC）of blank micelle （F127/TPGS），in vitro release behavior and preliminary stability of Hyp-F 127/TPGS were investigated ，and absorption characteristics of Hyp-F 127/TPGS were investigated by in situ unidirectional intestinal perfusion model. RESULTS The optimal preparation technology of Hyp-F 127/TPGS included F127-TPGS mass ratio of 2∶1，hydration time of 2 h，and Hyp amount of 9 mg. Results of three validation tests showed that the EE of Hyp-F 127/TPGS was （87.20±0.99）％，and the DL was （5.02±1.20）％，deviations from predicted values were 0.92％ and 2.39％. The micelles prepared by optimal technology were yellow ，clear and transparent solution ，with good Tyndall effect ；under transmission electron microscope ，they were spherical ，complete and evenly distributed ；the particle size was （15.02±0.16）nm， the PDI was 0.092±0.031，and the Zeta potential was （－6.67±1.47）mV. The CMC of F 127/TPGS was 21 μg/mL，Hyp-F127/ TPGS was stable after 4 weeks of storage at 4 ℃，and the cumulative release rates of Hyp-F 127/TPGS and Hyp control were （66.30±2.93）％（96 h）and（99.24±0.27）％（60 h），respectively. Hyp-F 127/TPGS and Hyp reference were absorbed in each intestinal segment ，and the main absorption sites were jejunum and duodenum respectively ；drug absorption rate constant andapparent absorption coefficient of the former were significantly higher than those of the latter （P＜0.05 or P＜0.01）. E-mail：zhangyuhangxz@163.com CONCLUSIONS The optimized preparation technology of Hyp-F127/TPGS is stable and feasible ；prepared Hyp-F 127/ TPGS shows a sustained -release effect ，which promotes the intestinal absorption of H yp to a certain extent.
ABSTRACT
OBJECTIVE： To establish the method for simultaneous determination of saikosaponin a and saikosaponin d in Bupleurum chinense water extract， and to optimize its water extraction technology for electromagnetic cracking. METHODS： HPLC method was used. The determination was performed on SB-C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 40 ℃. The detection wavelength was set at 210 nm， and the sample size was 10 μL. Based on single factor experiment， using extraction time， particle size， solide-liquid ratio as factors， total extraction rate of saikosaponin a to saikosaponin d as indexes， the extraction technology was optimized by using Box-Behnken response surface methdology， and compared with the results of ultrasound method and decoction method. RESULTS： The linear range of saikosaponin a and saikosaponin d were 50.70-202.80 μg/mL （r＝0.999 9） and 50.50-202.00 μg/mL （r＝0.999 9）， respectively. The quantitation limits were 0.16 and 0.13 μg/mL， respectively. The detection limits were 0.05 and 0.04 μg/mL，respectively. RSDs of precision， stability and reproducibility tests were all lower than 2％. The average recoveries were 98.23-102.47％ （RSD＝1.80％， n＝6） and 98.84％-102.06％ （RSD＝1.60％， n＝6）. The optimal extraction technology was as follows： the extraction time of 2.50 min； the particle size of 80 mesh， solid-liquid ratio of 1 ∶ 28 （g/mL）. Results of 3 times of validation tests showed that the optimal technology included the average total extraction rates of saikosaponin a and saikosaponin d were 8.42 mg/g， which was higher than that of ultrasonic method （8.34 mg/g） and decoction method （8.06 mg/g）， and the extration time was shorter. CONCLUSIONS： Established method is simple and accurate， and can be used for simultaneous determination of saikosaponin a and saikosaponin d in B. chinense water extract. The optimized water extraction technology for electromagnetic cracking is stable and feasible.
ABSTRACT
Objective:To study the biotransformation rule of the chemical components of total ginsenosides from ginseng stems and leaves tranformed by submerged fermentation of ganoderma lucidum,and to lay the foundation for further development and research of the medicinal fungus of ginseng stems and leaves.Methods:The total ginsenosides from ginseng stems and leaves were fermented by ganoderma lucidum,the changes of chemical components before and after fermentation were measured by thin-layer chromatography;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were detected by ultraviolet spectrophotometry;the levels of ginsenoside Rb1,Rg3,Rd,CK,Re,Rg,and Rh1 before and after fermentation were measured by high performance liquid chromatography.The human hepatocellular carcinoma SMMC-7721 cells were divided into blank control group,low,medium and high doses of total ginsenosides from ginseng stems and leaves groups and medicinal fungus of ginseng stems and leaves groups.The dosages of total ginsenosides from ginseng stems and leaves and medicinal fungus of ginseng stems and leaves were 5,10 and 20 mg·L-1,respectively.The inhibitory rate(IR) of the cells was determined by MTT assay.Results:The results of thin-layer chromatography showed that the saponins before and after fermentation occured mutual transformation;the levels of total ginsenosides from ginseng stems and leaves before and after fermentation were 749.98 and 602.26 mg·g-1,respectively;the levels of panaxadiol saponins Rb1,Rg3,Rd and CK before fermentation were 24.54,2.21,87.22 and 0 mg·g-1,and the levels of panaxatriol saponins Re,Rg and Rh1 were 151.34,77.02,and 3.06 mg·g-1.After fermentation,the level of Rb1 was decreased by 61.45%,the levels of Rg3,Rd and CK were increased by 238.91%,34.43% and 268.00%.The levels of Re and Rg1 were decreased by 63.75% and 64.41%,and the level of Rh1 was increased by 408.88%;the IR of the SMMC-7721 cells in medium dose of medicinal fungus of ginseng stems and leaves group was higher than that in medium dose of total ginsenosides from ginseng stems and leaves group(P<0.05);the IR of the SMMC-7721 cells in high dose of medicinal fungus of ginseng stems and leaves group was higher than that in high dose of total ginsenosides from ginseng stems and leaves group(P<0.01).Conclusion:The total ginsenosides from ginseng stems and leaves transformed by the submerged fermentation of ganoderma lucidum can significantly change its chemical components,which have stronger anti-proliferation activity in the SMMC-7721 cells.