Preparation Of Anti-hepatitis Effective Part Of Licorice And Its Nanoparticles

Hepatitis is one of the serious disease which threaten to human health, and in recent years Traditional Chinese Medicine has made great progress on the treatment of Hepatitis. The present invention relates to a process for the treatment of viral hepatitis by searching for effective fraction from Traditional Chinese Medicine. Leguminosae Glycyrrhiza licorice is commonly used in traditional Chinese medicine, which is composed of Triterpenoid saponins such as Glycyrrhizin and FGU major in LQ. Recently all sources mention its use for antispasmodic analgesic, anti-virus, anti-inflammatory, anti-cancer and other pharmacological effects, in addition, the treatment of hepatitis using licorice have been reported at home and abroad. The general anti-hepatitis drug is lack of targeting, while making use of nanoparticles as drug carrier can effectively accelerate the drug into the liver cells, in further enhancing liver targeting of drugs and efficacy. Thus it can be used to the treatment to viral hepatitis.This topic relates to a process for searching for an anti-hepatitis effective fraction from Licorice and the preparation of nanoparticles. First of all, we must make full control of the quality of licorice, provide a stable source of medicine for the preparation. Meanwhile we adopt high performance liquid chromatography to establish the fingerprint of Glycyrrhiza uralensis from different Fisch origin, and then extract from licorice and purify the effective parts of anti-hepatitis-total saponins from liquorice.. Furthermore, we study the process of the extraction and purification technology of total saponins from liquorice., use Carboxymethyl chitosan as drug carrier to prepare for nanoparticles,and make evaluation of its quality. The main contents and conclusions are as follows:1. To establish the HPLC fingerprint of Glycyrrhiza from different habitats. Separation was performed on a Hypersil ODS2 C18 column(4.6mm×250mm; 5μm) with methanol-0.1% phosphate water as mobile phase using gradient elution at flow rate of 0.6mL/min. The detection wavelength was at 254 nm and the analysis time was 70 min. The HPLC fingerprints of Glycyrrhiza with good separation were set up. And there were 11 common peaks. The results of method validation was consistent with the technical standard of fingerprints. The method of fingerprints was stable and reliable,so as to apply to the quality control of Glycyrrhiza. 2. To investigate the technological parameters of the extraction and purification process of total saponins from liquorice. Studied the extraction of total saponins from liquorice with orthogonal test; The technical process for purification of total saponins with the macroporous resins was screened by yield of total saponins as index. The optimum technological conditions for extraction:the water consumption 8 times, the total time 2h, the decoction times 3 times; The optimum parameters with resin HPD300 were established as followings:5 BV sample extract whose concentration was 0.2g (crude drug)/mL was passed through the column,and then the column was eluted with 6 BV 50% ethanol. With definite technical process, the total saponins from liquorice could be well extracted and purified.3. To prepare the nanoparticles of total saponins from liquorice and to evaluate its quality. The formulation of NP were optimized by the orthogonal test. The morphology was examined by the transmission electron microscope. The ultraviolet spectrophotometer was employed to determine the entrapment rate and drug-loading rate of NP. The diameter of NP were investigated by laser light scattering analysis instrument. And the release performance of NP in vitro was studied. The nanoparticles were round, smooth particles with average size of 114nm, entrapment rate 43.4% and drug-loading rate 16.4%. The result can provide a reference for studying the nanoparticles of total saponins from liquorice.