Preparation, Characterization, and In Vivo Evaluation of Amorphous Icaritin Nanoparticles Prepared by a Reactive Precipitation Technique.

Icaritin is a promising anti-hepatoma drug that is currently being tested in a phase-III clinical trial. A novel combination of amorphization and nanonization was used to enhance the oral bioavailability of icaritin. Amorphous icaritin nanoparticles (AINs) were prepared by a reactive precipitation technique (RPT). Fourier transform infrared spectrometry was used to investigate the mechanism underlying the formation of amorphous nanoparticles. AINs were characterized via scanning electron microscopy, X-ray powder diffraction, and differential scanning calorimetry. Our prepared AINs were also evaluated for their dissolution rates in vitro and oral bioavailability. The resultant nanosized AINs (64 nm) were amorphous and exhibited a higher dissolution rate than that derived from a previous oil-suspension formulation. Fourier transform infrared spectroscopy (FTIR) revealed that the C=O groups from the hydrophilic chain of polymers and the OH groups from icaritin formed hydrogen bonds that inhibited AIN crystallization and aggregation. Furthermore, an oral administration assay in beagle dogs showed that Cmax and AUClast of the dried AINs formulation were 3.3-fold and 4.5-fold higher than those of the oil-suspension preparation (p < 0.01), respectively. Our results demonstrate that the preparation of amorphous drug nanoparticles via our RPT may be a promising technique for improving the oral bioavailability of poorly water-soluble drugs.

[Genetic relationship and population genetic diversity of Epimedium pubescens and its related species based on ISSR and non-glandular hair characteristic].

In this study, 128 individuals form 14 Epimedium pubescens populations and 1 E. stellulatum population were analyzed by ISSR marker. The data were calculated by POPGENE software and clustered by UPGMA method. Optical microscope was used to observe the main types of the non-glandular hairs and their characteristics in each population. It is found that the following conclusions: Non-glandular hairs can be divided into five morphological categories, long straight pubescent, curly pubescent, appressed curly pubescent, pseudo short appressed hairs and long appressed. Eight primers were screening and a total of 94 bands were detected in ISSR, among which 90 were polymorphic bands. Based on the results of ISSR cluster analysis, 15 populations were divided into 3 clades. E. stellulatum populations should be incorporated into the E. pubescens or as avariety under E. pubescens not be independent and as it has no separate phylogenetic branch for a cluster. The genetic relationship among the populations of E. pubescens was closely related with its geographical distribution and non-glandular hair features. But there were also some inconsistency, which provided a good hint for the further study on the interspecific relationship and natural speciation manner of Epimedium species. Population diversity analysis showed Nm=0.354 4, Nei's=0.585 2. It was showed that E. pubescens has high genetic diversity among populations, for which the main reason was probably the high inbreeding rate and the small range of seed dispersal.

[Identification of Epmedii Folium and its counterfeit leaf of Quercus variabilis].

Epmedii Folium is a commonly used traditional Chinese drug, and is beneficial for the "liver" and "kidney" s function in Chinese medicine. Recently, the origin of this drug is more complex. Most of the identification studies are emphasized on the species certified by the pharmacopoeia and other related species from the same genus of Epimedium, but few was emphasized on the counterfeit. In this paper, one counterfeit of Epmedii Folium, identified as the dried leaf of Quercus variabilis (Fam. Fagaceae), has been reported based on field investigation, comparing specimen of Epmedii Folium and Q. variabilis,using the macroscopic, microscopic and TmC methods. It is resulted that they could be identified clearly not only by the macroscopic features, such as the vein character and the tooth apex, but also by the microscopic features, such as the vascular bundles of the midrib, the non-glandular hair, the anticlinal wall of the epidermis cell and the calcium oxalate crystal. Furthermore their TLC chromatograms showed also difference. This study will give reference for the identification of Epmedii Folium and the related supervision and inspection work.

Chemical pattern-aided classification to simplify the intricacy of morphological taxonomy of Epimedium species using chromatographic fingerprinting.

Epimedium herb (Yinyanghuo), one of the popular Chinese materia medica, is a multiple species colony of Epimedium genus belonging to Berberidaceae. There are five species of Epimedium that have been officially adopted in Chinese Pharmacopoeia under the same crude drug name 'Yinyanghuo' comprising Epimedium brevicornu, E. koreanum, E. sagittatum, E. pubescens, and E. wushanense. In addition, non-official species like E. acuminatum, E. miryanthum and E. leptorrhizum are also mix-used. Frequently, the morphological taxonomical identification is very difficult during on-site inspection for species authentication in the market. Researchers are often bewildered by the multiple species ambiguity when putting this crude drug in use. Referring to the bioactive constituents that are vital for therapeutic efficacy, the key to clarifying the multiple species confusion should rely on analysis of the bioactive composition. It is well known that medicinal Epimedium herbs contain special C-8 prenylated flavonol glycosides which contribute to various bioactivities and the major four, epimedin A (A), epimedin B (B), epimedin C (C) and icariin (I), are unanimously used as bioactive markers for quality control. In this study, HPLC-DAD fingerprinting was performed for investigating the molecular spectrum of various Epimedium species. It was found that the four major flavonoids constitute the middle part of the chromatographic profiles to form a specific region (named as 'ABCI fingerprint region') being dominant in the HPLC profiles of all medicinal Epimedium species, and the five official species express five different 'ABCI' patterns (different peak: peak ratios). Our study found that the convergent tendency of the 'ABCI region' among multiple species of Epimedium could facilitate differentiation of complex commercial samples based on similar bioactive composition should confer similar bioactivities. Merging the different species that possess the same 'ABCI region' pattern into the same group can create a simpler bioactive-fraction-aided classification array by clustering the commercial samples into three bioactive ingredients-based fingerprint patterns - 'E.b. pattern', 'E.k. pattern' and 'extensive E.w. pattern'. This approach offers the feasibility of characterizing and quality-controlling complex samples in the same genus designated under a single herbal drug entity on the premise of possessing the same bioactive ingredients pattern and supported by long-term traditional usage.

[Species and geographic distribution of large-flowered taxa of Epimedium in China].

OBJECTIVE: To introduce species and geographic distribution of large-flowered taxa of Epimedium L. in China. METHODS: Resources investigation and taxonomy study. RESULTS: There are 27 species large-flowered taxa of Epimedium in China. CONCLUSION: 13 species in Sichuan, 9 species in Hubei, 8 species in Guizhou, 4 species in Hunan, 3 species in Guangxi, 2 species in Chongqing, 2 species in Yunnan, 1 species in Jiangxi, 1 species in Shanxi, 1 species in Jilin, 1 species in Liaoning and 1 species in Heilongjiang.

[Comment on main species of herba epimedii].

OBJECTIVE: To comment on main resource species of Herba Epimedii. METHOD: Taxonomy, resource investigation and quality evaluation of flavonoid contents were made. RESULT: 7 species of Herba Epimedii having broad distribution were summarized. According the area of their distribution, they were named Epimedium sagittatum, E. brevicornu, E. acuminatum, E. koreanum, E. pubescens, E. leptorrhizum and E. wushanense. Each distribution in detail and icariin and likewise compounds contents of these 7 species were provided. Other 8 Epimedium species much used were proposed. Medicinal characters 15 species were provided in this paper. CONCLUSION: On the basis of distribution character, resource quantity and growing with other species mingled, it is suggested that 4 main production or purchasing districts in China be divided. The most important district is northeastern district, where only E. koreanum grows there, with the best guality. Northwestern and northern district is the second one, where E. brevicornu, mainly grows with much stable guality. Then it is eastern and southern district, where E. sagittatum grows with the largest quantity but unstable quality. Southwestern district is the last one, mingle species complex and identification important. In Sichuan province, E. acuminatum and E. pubescens are main species, with other 12 species growing there. In Guizhou province, E. acuminatum, E. leptorrhizum and E. wushanense are main species, other 7 species growing there and E. leptorrhizum with poor quality. It is suggested that E. acuminatum be recorded in Chinese Pharmacopoeia.