Study On Processing Detoxin And Technology Of Pinellia Rhizoma Prepared By Ethanol

This project was supported by national natural science foundation （No.30973939, No.81173549） and innovative projects of postgraduates in Jiangsu Province （CXLX11₀774）.Pinellia ternata （Thunb.） Breit, belonged to Araceae family, the tuberous stems of which are widely used in Chinese medicine, has the effects of drying dampness and dispersing phlegm, falling the versus and resolving phlegm, disintegrating lump and coalescence. P. Rhizoma is prepared before use to avoid its toxic irritantion to oral cavity and throat.P. Rhizoma is prepared to reduce irritation and alumen is one of the excipients which is often used in the process of preparing P. Rhizoma. The level of residual aluminum in P. Rhizoma is high after being prepared by alumen, and it is proven that intake too much Al3+will lead to Alzheimer’s disease and hypochromic anemia. Therefore, examination of the level of the alum residual amount of P. Rhizoma prepared by alumen, P. Rhizoma prepared by alumen and ginger are recorded in Chinese Pharmacopoeia （2010）. In addition, P. Rhizoma immersed in water or alum solution for a long time in the traditional processing, so the active ingredient is easy to loss, and it is also easy to cause moldy and smelly in summer because of its starch and polysaccharides soak long in high temperature condition. So far, any new effective method of reducing the toxicity of P. Rhizoma has not been found.Researchs show that the poisonous components of P. Rhizoma are toxic raphides, which are composed of mainly calcium oxalate crystals and protein. The irritation induced by raphides is due to the dual action of the sharp raphides in puncturing and PTA in causing inflammation. The raphides gradually lost the irritation when immersed in ethanol or prolonged heating.Wine is an excipient which commonly used to prepare Araceae in ancient times, whose main ingredient is ethanol. This paper was put forward the hypothesis "The toxicity of P. Rhizoma can be reduced by being immersed in ethanol and heating. The mechanism is ethanol and heating make the protein of P. Rhizoma denaturation, reduce the chemical damage of PTA, while destroying the crystal structure of the raphides to reduce the mechanical damage caused by raphides". This paper was designed according to this line of the research hypothesis.This paper examines the impact of temperature and solvent on P. Rhizoma, raphides and PTA, and research showed that ethanol and heating could reduce the irritation of P. Rhizoma and raphides, made the protein of P. Rhizoma denaturation, reduced the chemical damage of PTA, while destroying the crystal structure of the raphides to reduce the mechanical damage caused by raphides. The best technology was determined by orthogonal test L9（3）4and verified by the Draize rabbit eye test. Compare the differences in pharmacological effects and chemical composition of P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumen. The results showed that the P. Rhizoma prepared by ethanol is a reasonable feasibility. Main contents and findings were the following:1Study on the processing detoxin mechanism of P. Rhizoma prepared by ethanolStudy the influence of solvent and temperature on the irritation of P. Rhizoma and raphides by the Draize rabbit eye test. Observe the structure of raphides before and after being prepared by ethanol by SEM.SDS-PAGE electrophoresis were used to examine the influence of alcohol on the protein of P. Rhizoma and PTA. And study the impact of alcohol on the ability of PTA in causing inflammation. The results showed that ethanol and heating could reduce the irritation of P. Rhizoma and raphides, and destroy the raphides structure and crystal form;The chemical damage of PTA prepared by ethanol was reduced; The result of SDS-PAGE proved that that ethanol and heating could make PTA degeneration. Above results show that ethanol and heating could make PTA degeneration, reduce the chemical damage of PTA, and destroy the raphides structure and crystal form, reduce the mechanical damage.2Study on the technology of P. Rhizoma prepared by ethanolReduce the toxicity of P. Rhizoma by ethanol. The total free organic acid content was chosen as a chemical index and the PGE2of rats’abdominal cavity was chosen as a toxicological index. The amount of ethanol, preparing time, ethanol concentration and preparing temperature were investigated. The best technology was determined by orthogonal test L9（3）4and verified by the Draize rabbit eye test. The optimal technology was that P. Rhizoma was prepared by75%ethanol at the temperature of40℃by4days, and then dried. Compared P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumen time required at room temperature. It was proved that P. Rhizoma prepared by ethanol need10days, P. Rhizoma prepared by alumen need7days in the spring and winter（10℃）; P. Rhizoma prepared by ethanol need6days, P. Rhizoma prepared by alumen need5days in the summer and autumn （30℃）. All of them could reduce the irritation on the rabbit conjunctiva. The time of prepared P. Rhizoma by ethanol is little longer than prepared P. Rhizoma by alumen under the same conditions. The calculated rate found for P. Rhizoma prepared by ethanol is higher than P. Rhizoma prepared by alumen. In summary, P. Rhizoma prepared by ethanol is simple, feasible, and can significantly improve the production environment, and avoid the intake of aluminum ions. It can be further derived the new method to prepare P. Rhizoma is feasible.3Comparative study on the efficacy of P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumenThe cough relieving and sputum elimination effects of P. Rhizoma prepared by ethanol were observed by the ammonia water-induced tussive mice and excretion of phenol red in the airway of mice, and the anti-inflammatory effect was evaluated by the models of dimethylbenzene-induced mouse ear swelling. It was proved that the effects of relieving cough, reducing sputum and anti-inflammatory of P. Rhizoma were not reduced after being prepared by ethanol.4Comparative study on the content of extract in P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumenCompare the content of water-soluble and alcohol-soluble extract in P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumen, to examine the rationality of the optimal technology. The results showed that the content of water-soluble and alcohol-soluble extract of P. Rhizoma declined slightly after being prepared by ethanol, while reducing by nearly half in P. Rhizoma prepared by alumen.5Comparative study on the content of total free organic acid in P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumenPotential residual titration was used to determine the total free organic acids in P. Rhizoma, and compare the content with P. Rhizoma prepared by ethanol、P. Rhizoma prepared by alumen. The results showed that the content of total free organic acid in P. Rhizoma prepared by ethanol was similar to P. Rhizoma, while the content in P Rhizoma prepared by a lumen was higher.6Comparative study on the content of five nucleosides in P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumenDetermined and compared the content of Uracil, Xanthine, Uridine, Carnine and Guanosine in P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumen.The total content of five nucleosides in P. Rhizoma was0.06158%, and fell to0.03242%、0.00350%after being prepared by ethanol and alumen respectively, decreased by47.25%and94.32%. The method of preparing P. Rhizoma by ethanol was better at the aspect of keeping nucleosides.7Compare the fingerprint of P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumenUracil, Xanthine, Uridine and Guanosine were chosen as comparing peaks, compare the fingerprint of P. Rhizoma、P. Rhizoma prepared by ethanol and P. Rhizoma prepared by alumen.17fingerprint peaks in common were confirmed in P. Rhizoma, the number fell to15and12after being prepared by ethanol and alumen respectively. The fingerprint peaks of P. Rhizoma didn’t change after being prepared by ethanol, while the number and area of the fingerprint peaks reduced dramatically after being prepared by alumen. The method of preparing P. Rhizoma by ethanol was better at the aspect of keeping the water-soluble components.After studying on the processing detoxin mechanism of P. Rhizoma prepared by ethanol, establish the best technology, alternative the method of prepared by alumen.