Studies On Reducing The Toxins Of Qingmuxiang (Aristolochia Debilis) Containing Aristolochic Acids By Processing Technique

Aristolochic acid (AA) are commonly found in medicinal plants such as radix aristolochiae. AA is nephrotoxic and carcinogenic, and have been reported to involve in a specific type of renal fibrosis and end-stage renal failure. It also cause acute hepatitis after long time administration. It dues to the toxicity that confined the use of this kind of Chinese herb. Our research group brought forward the new processing method of removing toxicities and reserving pharmacological effects from the herbal medicine containing AA at the same time, which was supported by National Fundation of Natural Sciences in 2002 (No. 30271612). The research work was to reduce the toxins, aritolochic acids, of some Chinese herbal drugs by Chinese unique processing technique, and hold their pharmacological effect at the same time. The methodological studies perhaps offer a selection for Chinese herb to reduce toxins, to guarantee the safety for clinical uses.We had made use of this processing method to Qingmuxiang, stem of Aristolochia debilis Sieb.et Zucc. Both investigation in the Processing standardization and toxicological evaluation indicate that AAs in A. debilis can be removed mostly off by this processing method and the toxicities were largely reduced to the designed objective, compared with that of raw material. But the efficiency of processing product and raw material were doubtful of diuresis effects. The optimized techniques were evaluated by studies on reducing the toxins and on holding pharmacological effects.During the studies on reducing toxin, we firstly established a method to determine the contents of aristolochic acid A in Aristolochia debilis. Secondly, the processing techniques that developed by orthogonal designed methods was to dip the crude drug material in alkali water and then to fry it with vinegar. The removed ratio of toxins were above 81% by three batches medium-scale production. Thirdly, the acute toxicityof processing product was decreased 4.5 times compared with that of raw material in the acute toxicity evaluation. Finally, the common behavior, blood biochemical indexes, urine indexes, viscera coefficient tissue pathology and electrocardiogram were observed in the three month long-term safety evaluation. All these results indicated that the toxicity of the processed product were much lower than that of the raw material.During the studies on holding pharmacological effects, basing on the clinical research of qingmuxiang, we chose the three indexes on restrained intestinal squirm, analgesic and anti-inflammation to study the pharmacological effects of raw Qingmuxiang and processed qingmuxiang respectively. The experimental data indicated that the processed qingmuxiang did hold the pharmacological effects.To sum up, the processing method combining basification and acetification could reduce the content of toxic arisolochic acid A in qingmuxiang. This thesis demonstrated from chemistry, analysis, pharmacology and toxicology studies that the processing method could reduce the toxicity and hold the efficacy at the same time.This thesis summarizes the correlated investigation ,including bencaological study,confusing species, physics and chemistry discrimination,chemical composition and contents, traditional processing condition, pharmacological studies, clinical application, toxic and adverse action, and so on.We optimized the techniques of processed Qingmuxiang and determinated the contents of AA during different time in alkali water.The removed ratio of toxins are 95.87 % in former method while those are 99.38 % in this newly optimized techniques. Studies indicated that pure water makes for reducing the contents of AA and ions of Natrium. The removed ratio of toxins were 24.00 % in traditional method.We compared the raw Qingmuxiang with the processed one in LC-MS and identify four chromatogram peaks in the raw Qingmuxiang. They were aristololactam-N-P-D-glucopyranoside, Aristolochic acid C , 7-hydroxy-aristolochic acid A, aristolochic acid B. The components containing Aristolochic acids couldn't be identified from HPLC profile of the processed Qingmuxiang.We established a determination method of total aristolochic acid byultraviolet spectrophotometry. The total aristolochic acid of raw and processed Qingmuxiang were 0.543 % and 0.120 % respectively. We determined the contents of total aristolochic acid A. in debilis from different areas. The highest was 0.567% from Hebei Anguo, the lowest was 0.145% from Chongqing. We formulated the quality standard of the processed Qingmuxiang.We detected the contents of aristolochic acid A in Guanxisuhe pills and capsules from different factories by using RP-HPLC.The contents of aristolochic acid A in Guanxisuhe pills and capsules from different factories varied greatly. The highest is 0.516 mg'g"1 from Sanjiu group Co., and the lowest is 0.148 mg'g1 from Beijing Tongrentang Co-limited company.Otherwise, to generalize the correlative studies of Inula helenium L. and to compare Aristolochia debilis with them.Additionally, we separated the alkaloids from the raw Qingmuxiang and four compounds which had high chromatogram peaks in HPLC profile from the processed Qingmuxiang in order to reveal the processing priciple. Two structures had been confirmed of the separaed compounds. One was magnoflorine from the raw material and the other was Aristolenone (Al(10)Aristolenone-2) from the processed one.