The. Hyoscyameae Processing Technology And Quality Standards

Objective:Hyoscyamus niger L., originally called"LangDangzi", is the dried and mature seeds of Hyoscyamus niger L.. It is hypertoxic with the effects of relieving spasm and pain, relieving uneasiness and fixed pain, mainly used for the treatment of gastrospasm, dyspnea with coughing, vecordia, anti-shock and improving microcirculation. In recent years, we have further found its multilateral pharmacological effects and clinical functions. However, no parameter of procedure and quality control method is regularized in Pharmacopoeia of the People's Republic of China.And the ancient documents all recoreded many processing methods. In order to optimize the quality standard of Hyoscyamus niger L., it is necessary to do some further study on processing technology for this herbal plant, so as to found a proper technology that may reduce toxicity and enhance effect, be steady , controllable, easy to popularize and fit for mass production. At last, according to the influence of processing on the active constituents, we optimized parameter of processing technology, formulated the reasonable processing method for the seeds of Hyoscyamus niger L. and established the quality control standard of processed products. Thus the study provids the assurance for the safety and efffectiveness of this medicinal.Material and method:Material:Seeds of Hyoscyamus niger was purchased in Anhui Chinese herbal medicines market, it has been identified as the dry mature seeds of Hyoscyamus niger L.by Prof. ZHAI Yan-jun in liaoning University of TCM.Method:1. Used a series of extracts to perform MTT assay to study the action of the medicinal plant on the cultured human lung cancer cell proliferation.2. The chemical constituents were isolated and purified by various chromatographic methods including silica gel column chromatography, Sephadex LH-20 column chromatography and HPLC. Their structures were identified by the physical-chemical properties and spectra data.3. The variation of chemical constituents in the crude and processed medicinals were comparatively analysed by means of UV, HPLC and ICP-MS.4. The reasonable process parameters were optimized and the best processing technology was founded by orthogonal experiment.5. The contente of active constituents in the raw and processed product were determined by HPLC. microscopic identification,TLC,moisture content,ash,and acid-insoluble ash etc. were also investigated. The quality standard of processed product was established.6. The gradient elution method for HPLC fingerprint was established to estimate the quality of the raw and processed medicinals.Result1. It was howed that the extract of some petroleum ether, chloroform and ethyl acetate part have different degrees of inhibition on lung cancer cells , the inhibition being dose dependent.2. From the extracts of the ethyl acetate part of the medicinals,8 compounds were isolated and identified. Among them, there were 2 ith anolides and 4 lignans.3. There were significant differences in the content of chemical constituents between the raw and processed medicinals, also among various habitats.4. The optimal processing technology was set: taking Hyoscyamus niger seeds,cleaning, setting in a frying container and stir-frying at 110～120℃for 4 minutes.5. The limit of moisture,ash and acid-insoluble ash of Hyoscyamus niger seeds were temporarily decided as no more than 10.0%,8.0%,3.0% respectively. Alcohol-soluble extract was no less than 15.0%. The results of active constituents all meeted with the related standards.6. There was a difference in HPLC-fingerprints between the raw and processed, but the samples from various habitats were of high similarity, and only a few samples had slightly difference.Conclusion:1. Seeds of Hyoscyamus niger extracts have anti-tumor effects in vitro. Hyoscyamus niger extract has a lantency of developing anti-tumor action. 2. It is first time to analysis the compound of the ethyl acetate part of the sample, compound6 is the first time isolated in this species and it is also a new structure.3. The variation of chemical ingredients between the raw and processed medicinals was analysed and the impact of processing on the chemical constituents was studied by means of chromatographic methods.4. The optimum processing technology of stir-frying Hyoscyamus niger seeds was stable and feasible.5. The quality standard of the raw and processed Hyoscyamus niger seeds has been established.6. The RP-HPLC fingerprint method is repeatable, feasible in analysis of Hyoscyamus niger seeds and can be used in quality assessment of Hyoscyamus niger seeds.The chemical components in the raw and processed seeds of Hyoscyamus niger are significantly different but no difference among those in various habitats.