Study On The Influence Of Quality In Smilax Glabra Roxb.by Sulfur-fumigated Process

The rhizomes of Smilax glabra Roxb.?Liliaceae?have been extensively used in Chinese traditional medicines,called“Tufuling”for the treatment of syphilis,acute bacterial dysentery,acute and chronic nephritis.On the basis of previous studies,we studied the chemical composition,antioxidant activity and safety of Smilax glabra Roxb.before and after sulfur fumigation,aiming to provide scientific basis for quality evaluation and development and utilization of Smilax glabra Roxb.?1?The experiment found that fresh medicinal herbs and sulfur fumigation of medicinal herbs appearance and smell are significantly different,fresh herbs hard texture,the color of the cross-section after cutting is white,dried herbs cross-section of brown to reddish brown,with a special soil smell.After the fumigation of sulfur medicinal material texture is not hard,the color is pale golden yellow,and just fumigation herbs have pungent sulfur smell,and often accompanied by a special sour after drying.?2?To establish a method of determination seventh steroidal saponins content in Smilax glabra Roxb.by UPLC-DAD with a ACQUITY UPLC HSS C18?2.1mm×100mm,1.8?m?column,and gradually diluted with a 0.1%phosphoric acid water?A?-cetonitrile?B?at a flow rate of 0.2 mL·min^-1,elegant tube temperature35°C,gradient elution program:5-15%B?0-2 min?,15-20%B?2-5 min?,20-25%B?5-8 min?,25-30%B?8-10 min?,30-35%B?10-12 min?,35-40%B?12-15 min?,40-70%B?15-20 min?.The experimental results show that S-SGR compared with the SGR,the seven chemical components have declined,so the sulfur fumigation will reduce the effective content of Smilax glabra Roxb.,followed by the possibility of reducing its efficacy.?3?The antioxidant capacity of SGR and S-SGR sample extracts was evaluated.First,the content of total flavonoids in SGR and S-SGR crude extracts was determined by UV-visible spectrophotometry.The experimental results showed that the content of total flavonoids in S-SGR decreased by 19.14%compared with that of SGR,so the sulfur fumigation would affect the total flavonoids Content.?4?The SGR and S-SGR extracts were assayed for DPPH,ABTS⁺free radical scavenging ability and Fe³⁺reducing ability.The results showed that the IC50 value of ABR radical scavenging by SGR was 0.00402 mg·mL^-1,the IC50 value of scavenging ABTS radical by S-SGR was 0.00745 mg·mL^-1,and the IC50 value of scavenging DPPH radical by SGR was 0.00791 mg·mL^-1.The IC50 value of scavenging DPPH radical by S-SGR was 0.01695 mg·mL^-1.The reducing ability of S-SGR to iron ion was also weaker than that of SGR.The results show that sulfur fumigation Smilax glabra Roxb.its antioxidant capacity was significantly reduced.?5?The toxicity of SGR and S-SGR aqueous extracts were tested by acute toxicity test in mice.The experimental results showed that although the LD50 of S-SGR was not determined,we observed the appearance signs,general activity,S-SGR was found to be of low toxicity and to be used as a bioassay for testing of body weight and blood biochemical markers ALT,AST?IU/L?,DBIL?IU/L?,TBIL?IU/L?Mice can cause loss of appetite,weight loss,dry stools,darker colors,slowness of movement,elevated blood biochemical markers,and liver damage in mice.This indicates that the acute toxicity of Smilax china may increase after fumigation,but the detailed long-term toxicity needs further study.?6?The method of UPLC-QTOF-MS was established for the rapid identification of Smilax vulgaris smoked products.The chromatographic conditions were as follows:ACQUITY HSS T3?2.1×100 mm 1.8 um?;mobile phase:0.1%,Gradient elution5-10%B?0-1.0 min?,10-12%B?1-2.5 min?,12-25%B?2.5-7.0 min?,25-35%B?7.5-11.5 min?,48-70%B?11.5-12.0 min?,70-90%B?12.0-13.0 min?,90-5%B?13.0-13.1 min?,5%B?13.1-15 min?.Flow rate:0.5 mL·min-1,column temperature:35?,injection volume:2?l.The experimental results showed that among the 68compounds detected,58 compounds could be initially characterized.Six sulfur-containing chemical constituents of Smilax china that were fumigated by sulfur were analyzed by PCA and OPLS-DA to identify the fumigates Markers such as glucosylbutyric acid sulfates,5-O-coffee shikimic acid sulfites,3-O-coffee shikimic acid sulfites,5-O-coffee shikimate acids,Sulfate,3-O-Coffee shikimic acid sulfate,Astilosulfate,this method has been successfully applied to the rapid identification of sulfur fumigation of Smilax glabra Roxb.