| Radix Polygoni Multiflori, rhubarb and semen cassia were used as the research object, over the modern detection means, such as HPLC, HPLC- IT/MS,UPLC- QQQ/MS and UPLC-Q-TOF/MS, the main active components, anthraquinone compounds were in the qualitative and quantitative study in vivo and in vitro, combinning with present research status of anthraquinones, we can know the current situation in-and outside of the body systematically.(1) with one kind of method for the test sample preparation, 10 raw radix polygoni multiflori and 20 prepared radix polygoni multiflori, 11 rhubarb and 8 group of semen cassia have been material content determination.The repeatability of preparation method in range of 1.1 ~ 3.5%, the recovery rate for the main index from 95.31% to 107.06%, and both can get good stability within 24 h. The anthraquinones content of radix polygoni multiflori were found in difference markedly, and the anthraquinone content is about 0.25% for highest content, the lowest is only 0.01%. The anthraquinones content in most radix polygoni multiflori are less than that requied in Chinese pharmacopoeia 2010. The anthraquinones in all rhubarb samples can reach relevant provision on Chinese pharmacopoeia 2010, each batch of anthraquinones content were similar, and also have no significant difference between single anthraquinone ingredient, and the content of rhein is the highest. The two semen cassia batches,fome India, the content of chrysophanol is low and do not conform to the Chinese pharmacopoeia 2010 in the regulation.There are obvious difference in anthraquinone type and content for the three kinds of herbs,among them,the content of anthraquinones in rhubarb is maximum,the anthraquinones in semen cassia are various and nor do both in radix polygoni multiflori.(2) The structural analysis of the combined anthraquinone radix polygoni multiflori. Based on several combined anthraquinone reference substance,which is made after separation and purification, HPLC/IT/MS method is applied to qualitative analysis of the combined anthraquinone, and now emodin- 8-O-beta-D-pyranglucoside and physcion- 8-O-beta-D-pyranglucoside are identified.(3) The use of UPLC- QQQ/MS for analysis of anthraquinone ingredient metabolism in the body.Give the extractive of rhubarb and Radix polygoni multiflori with same anthraquinone content to SD rats, the former crude drug dosage of 10 g/kg rats and can be detected three components, the later for crude drug dose for 21 g/kg rats can be detected three target composition, emodin and emodin-8-O-beta-D- pyran glucoside were detected in both groups, by comparing the two components of blood drug concentration and drug parameters found that the absorption for emodin and emodin- 8-O-beta-D-glycosidase in Radix polygoni multiflori are faster than the rhubarb group, but its elimination is also faster than the rhubarb group. The other components in Radix polygoni multiflori and rhubarb affect the absorption and metabolism of emodin and emodin-8-O- beta-D-pyran glycosidase.(4) The use of UPLC- Q- TOF/MS for analysis of metabolism of anthraquinone in liver microsome incubation system. Emodin-8-O-beta-D-pyran glucoside, physcion-8-O-betaD-pyran glucoside, aloe-emodin, rhein, emodin, chrysophanol and physcion were used as substrate, incubating in liver microsomes from rat, mouse, dog and human. Oxidation reaction, such as the CH3-- CH2OH-- COOH oxidation conversion step by step inâ… .Six anthraquinones substrate have glucosealdehyde acid metabolites except physcion in â…¡.The metabolism of some kinds of anthraquinone monomer have obvious differences between species.(5) The selected three kinds of traditional Chinese medicine were made extraction process optimization by the method of dynamic optimization. Using dynamic optimization method of three herbs aqueous extraction process to determine the following: tenfold mass of water was added and then boiled for three times,and extracting time is respectively: 90, 70, 50 min for Radix polygoni multiflori; tenfold mass of water was added and then boiled for three times,and extracting time is respectively: 50, 50, 60 min for Rhubarb; tenfold mass of water was added and then boiled for three times,and extracting time is respectively:50, 50, 40 min for Semen Cassia.The extraction process was used to make preparation for long-term toxicity experiment. |
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