ABSTRACT
ObjectiveTo analyze the effects of new integration processing method in producing area and traditional method on the composition and pharmacological action of Polygoni Multiflori Radix Praeparata（PMRP）， and to illustrate the advantages of toxicity reducing and efficacy enhancing of the decoction pieces prepared by the new method. MethodFresh Polygoni Multiflori Radix（PMR） was taken from Dao-di producing area， and was processed by new integration processing method in producing area（steaming with black bean juice under pressure of 0.1 MPa and temperature at 120 ℃ for 10.5 h） and traditional method（steaming with black bean juice under water for 36 h）， respectively. Samples were collected during the processing process of the two methods， For new method， the samples were collected at 0.5， 3， 5.5， 8， 10.5 h， separately. For traditional method， the samples were collected every 4 h. High performance liquid chromatography（HPLC） was used to establish fingerprint and identify common peaks， the content of polysaccharides was determined by anthrone-sulfuric acid colorimetry at 627 nm， and the contents of anthraquinones and stilbene glycosides in different processed products were determined according to the methods under the item of determination of PMR and PMRP in the 2020 edition of Chinese Pharmacopoeia. In pharmacological experiments， 90 SD rats were randomly divided into 9 groups with 10 in each group（half of male and half of female）， including the blank group， and raw products， 24 h processed products under atmospheric pressure， 30 h processed products under atmospheric pressure， 8 h processed products under high pressure groups with low and high dosages（4.125， 16.5 g·kg-1）. Rats were given the drug by gavage for 29 d with once a day， blood was collected from the abdominal aorta after the last administration， and the serum was isolated， the body mass and liver mass of rats were weighed and the organ index was calculated. The pathological change of liver tissue was observed by hematoxylin-eosin（HE） staining， and biochemical methods were used to detect the contents of aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， alkaline phosphatase（ALP）， γ-glutamyltransferase（GGT）， lactic dehydrogenase（LDH） in serum which used as liver function indicators and the levels of superoxide dismutase（SOD）， malondialdehyde（MDA）， glutathione peroxidase（GSH-Px） in brain tissues which used as oxidation indicators. ResultA total of 14 common peaks were identified in the fingerprint of PMR， PMRP prepared by new method and traditional method， and three of the peaks were designated as stilbene glycoside， emodin and emodin methyl ether， respectively. The characteristic peak areas of each processed products changed significantly from 0 min to 25 min， indicating that different processing methods had an effect on the contents of components with high polarity in PMRP， and the trend of the changes of the two methods was similar， with the higher degree of change in the new method. The determination results showed that compared with the traditional method， the content of polysaccharide（a kind of beneficial component in PMRP obtained by the new method） significantly increased， while the contents of stilbene glycoside and bound anthraquinone（liver-damaging ingredients） significantly decreased. The pharmacological results showed that compared with the blank group， AST and LDH levels of male rats in the low and high dose groups of 24 h processed products under atmospheric pressure and AST level of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly reduced（P<0.05， P<0.01）， while compared with the raw product groups with the same dose， AST and LDH levels of male rats in the low dose group of 30 h processed products under atmospheric pressure were significantly reduced（P<0.05， P<0.01）， the AST levels of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly decreased（P<0.01）， and there was no statistical significance in the differences of biochemical indexes of female rats in each administration group as compared with those of the blank group. ConclusionThe new integration processing method in producing area of PMRP can reach the quality of relevant regulations in 8 h. The processed products obtained by this method have more advantages than the traditional method in terms of toxicity reducing and efficacy enhancing， and energy saving to avoid the loss of ingredients， which can provide ideas for the production of high-quality decoction pieces of PMRP， and the integration processing method in producing area of other roots and rhizomes of traditional Chinese medicines.
ABSTRACT
In order to ef fectively control the quality ofSophora lfos carbonisatus (flower and flower buds), this study established quality control methods and standard of the decoction pieces. Referring to the related methods in appendix of Chinese Pharmacopoeia (2010 Edition), the moisture, total ash, acid insoluble ash, alcohol extracts ofSophora lfos carbonisatus were measured, respectively, with rutin, quercetin as control substance. The eluents for rutin and quercetin are ethyl acetate - formic acid - water (8: 1:1) and chloroform - methanol - water (6.5:1:1), respectively and all TLC plates were observed at 365 nm. Total flavonoids are measured by visible - UV - spectrophotometric, and rutin and quercetin were determined by HPLC. The chromatographic conditions for rutin are: Kromasil C18 as the stationary phase, methanol -1% acetic acid (32:68) as mobile phase, flow rate: 0.8 mL·min-1, detection wavelength 257 nm,the column temperature 35℃; for quercetin: Kromasil C18 as the stationary phase, methanol -0.4% acetic acid (44:56) as the mobile phase, flow rate: 0.8 mL·min-1, detection wavelength 257 nm, the column temperature 40℃.The contents of moisture, total ash, acid insoluble ash, should not exceed 6%, 16%, 8.0% in flower and not exceed 6.0%,9.0%, 1.5% in buds, respectively. Under the conditions of TLC, in flower and flower buds, 2 reference substances can be separated well with others. Extract, total flavonoids, rutin, quercetin were no lower than 40.0%, 5.0%, 2.5%,0. 2% in flower and no lower than 45.0%, 10.0%, 5.0%, 0.9% in buds, respectively. The established standards can improve the levels of quality control, provide experimental data for safety and efficacy of clinical application of Sophora flos carbonisatus, and also offer supporting data for the Chinese Pharmacopoeia 2015 Edition.