Establishment of HPLC Fingerprint ，Multivariate Statistical Analysis and Processing Technology of Different Suet Oil-baked Epimedium brevicornum / 中国药房

OBJECTIVE：To establish HPLC fingerprint of different products of suet oil-baked Epimedium brevicornum ，and to screen the optimal baking technology. METHODS ：HPLC method was adopted. Using icariin as reference ，HPLC fingerprints of 22 batches of samples were drawn. The similarity was evaluated by using Similarity Evaluation System of TCM Chromatographic Fingerprint（2012 edition），and common peaks were confirmed. HCA ，PCA and OPLS-DA analysis were performed by SIMCA 14.1 statistical software. Taking variable importance in the project ＞1 as criteria ，biomarkers affecting the quality difference of suet oil-baked E. brevicornum were screened ；using mass marker as index ，the baking technology was screened by baking technology. RESULTS：There were 18 common peaks in 22 batches of samples. The similarities were between 0.831 and 0.991. Totally 7 common peaks were identified ，i.e. epimedin A ，epimedin B ，epimedin C ，icariin，sagittatoside A ，sagittatoside B ，baohuoside Ⅰ. The 22 batches of samples were clustered into two categories ，S19-S22 were clustered into category Ⅰ and S 1-S18 were clustered into category Ⅱ. The category Ⅱ was sub-clustered into category Ⅱa（S15-S18），category Ⅱb（S10-S14），category Ⅱc （S1-S9）；the result of PCA analysis was consistent with above results. OPLS-DA showed that the biomarkers affecting the quality difference were icariin ，sagittatoside B and baohuoside Ⅰ. The results of kinetic studies showed that the content of icariin when baked at 180 ℃ for 25 min or 190 ℃ for 20 min，that of baohuoside Ⅰ when baked at 180 ℃ for 30 min or 190 ℃ for 15 min and that of epimedin B when baked at 210 ℃ for 18 min were the highest ；according to above results ，the optimal baking technology was baking at 180 ℃ for 25-30 min or 190 ℃ for 15-20 min. CONCLUSIONS ：Established fingerprint is stable ， reliable and reproducible. The multivariate statistical analysis can be used for the changes of chemical components in E. brevicornum under different baking condition and preliminary selection of baking technology.

Effect of processing excipient suet oil on formation and absorption of baohuoside Ⅰ-bile salt self-assembled micelles / 中国中药杂志

The fried method with suet oil,which can strengthen the effect of Epimedium in warming kidney and enhancing Yang,has been widely used in the processing of Epimedium in traditional Chinese medicine. Based on the formation mechanism of Epimedium flavonoids self-assembled micelles in vivo,the synergistic mechanism of processing excipient suet oil was investigated in this paper from the perspective of pharmaceutics. Baohuoside Ⅰ,as representative component of processed Epimedium,was selected as model drug.Average size and zeta potential were measured and the morphology of micelles was observed under transmission electron microscopy. Caco-2 monolayer cell model,rat intestinal perfusion model and in vivo serum drug concentration method were established to investigate the effect of suet oil on the formation and absorption of the baohuosideⅠ bile salt self-assembled micelles. Baohuoside Ⅰ can form selfassembled micelles under the action of sodium deoxycholate. While,adding suet oil into the baohuoside Ⅰ-bile salt micelles( BSDOC) can make it form a more stable system with a smaller average size,higher Zeta potential,lower polydispersity index( PDI) value,significantly improved encapsulation efficiency and drug loading,indicating that suet oil could significantly improve the micelle formation in vivo. In addition,the permeability coefficient of baohuoside Ⅰ in Caco-2 monolayer cells and the four intestinal organs( duodenum,jejunum,ileum and colon) was increased and the oral bioavailability was also improved after adding the suet oil to BS-DOC.All the results demonstrated that the suet oil can promote the formation and absorption of baohuoside Ⅰ self-assembled micelles,so as to enhance its synergistic effects.

Processing Study of Epimedium Flavonoids Self-Assembled Micelles Fried with Suet Oil on Its Bioavailability / 中国药学杂志

OBJECTIVE: To study the influence of bioavailability of Epimedium flavonoids after processing by suet oil. METHODS: Pharmacokinetics parameters of rats Epimedium flavonoids and icariin and baohuoside I was determining by the method of pharmacology effect and plasma concentration method. RESULTS: The concentration of the Epimedium flavonoids, icariin and baohuoside I in micelles groups were higher than that of the active pharmaceutical ingredient(API). ρmax were increased by 77.94%, 38.99% and 49.90%; AUC0-t increased by 95.19%, 37.76% and 43.49%; AUC0-∞ increased by 119.35%, 35.37% and 46.26% on Epimedium flavonoids, icariin and baohuoside I group. CONCLUSION: Suet oil improved the bioavailability of Epimedium flavonoids and promoted its absorption.

Synergistic effect of Epimedii Folium fried with suet oil for warming kidney and enhancing yang in dosage form of self-assembled micelles / 中国中药杂志

The fried method with suet oil, which can strengthen the effect of Epimedii Folium in warming kidney and enhancing yang, has been widely used in the processing of Epimedii Folium in traditional Chinese medicine. In this paper, we adopted hydrocortisone-induced kidney-yang deficiency rat models, and took physical signs, serum testosterone, corticosterone levels and histopathological examination of rats as indictors to systematically study the synergistic effect of suet oil in warming kidney and enhancing yang of fried Epimedii Folium in the dosage form of self-assembled micelles. Drugs were given orally, and compared with model group, indicating normal physical signs and biochemical indicators, and significant alleviation in disorder of hypothalamic-pituitary-adrenal-thymus (HPAT) axis suppression. All these experimental results indicated that both Epimedium flavonoids and its self-assembled micelles, especially the processed self-assembled micelles of Epimedii Folium, showed the significant effects in warming kidney and enhancing yang, which might be correlated with the regulation of the disorder of HPAT axis suppression and the improvement of major pathological conditions of testicular gonadal tissues. This discovery can further reveal the synergistic effect and mechanism of processing material suet oil on Epimedii Folium.

Promotion of suet oil to absorption and transportation of total flavonoids from Epimedii Folium / 中草药

Objective: To study the promotion mechanism of suet oil to the intestinal absorption of the total flavonoids from Epimedii Folium (EF). Methods: The in vivo intestinal perfusion model and Caco-2 cell monolayer model of rats were used to study the influence of processing excipient suet oil in self-assembled micelles on the intestinal absorption of total flavones from EF. Results: In the in vivo intestinal perfusion model of rats, there were differences of absorption of icariin in the total flavonoids from EF in the four segments of the intestines. After the suet oil was added and self-assembled micelles were formed, the permeabilities of icariin in the total flavonoids from EF were increased significantly in duodenum and jejunum segments. In Caco-2 cell monolayer model of rats, the absorptive permeability coefficients were small for icariin in the total flavonoids from EF, after the suet oil was added and the self-assembled micelles were formed, the absorptive permeability coefficients were increased significantly, the efflux ratios were decreased from 4.72 to 2.31. Conclusion: The total flavonoids from EF have a bad intestinal absorption, after the suet oil added and the self-assembled micelles formed, the intestinal absorption of the total flavonoids from EF could be improved.