| Objective To excavate the information about Tripterygium wilfordii,including its alias,morphology,nature,taste,meridian tropism,growth environment,main diseases,adverse reactions and main active ingredients in ancient books and modern literature,and to analyze the clinical application and detoxification measures.To develop a novel extraction method to rapid screening the antioxidants in Tripterygium wilfordii Hook.F(TWHF).This might be a new strategy for screening the antioxidants in related production.To build up quantification method for these antioxidants which might be potential quality control method for Tripterygium wilfordii Hook.F.It will be benefit to further investigation or application of this medicine in different areas.Methods Literature research:Chinese medical records(ZhongHuaYiDian),Han Tong ancient books,Ding Xiu ancient books database,CNKI,Wan Fang,VIP database,Pubmed and Embase English database were selected as objects of literature retrieval.By using the method of computer search and manual search to screen the literature,this paper summarized the alias and morphology of Tripterygium wilfordii,nature,taste,meridian,growth environment,medicinal parts,toxic and side effects and treatment measures.Through Excel 2016 software,the data of indications,toxic and side effects and effective components of Tripterygium wilfordii were statistically analyzed.The experimental research:QuEChERS method was first applied in extraction of antioxidants in TWHF.The parameters which may affect the extraction efficiency was optimized,including extraction solvent,extraction buffers,salts used in salting out process,extraction time and solid sorbents.The peak area in HPLC was selected as key factor to evaluate the extraction efficiency.Four major antioxidants were further identified by LC-MS.The final structure information was confirmed by comparison with reference standards.Besides,antioxidant assay was applied to the QuEChERS extract and these four components.As in the salting out process,non-volatile salts were used,this may cause contamination in interface of mass spectrometry.Ultrasound-assisted extraction method was developed to extract these four antioxidants.The extraction parameters,such as extraction solvent,solid-liquid ratio,extraction time,extraction temperature was investigated.The content of these four components in seven different producing areas was quantified by LC-MS method.Statics analysis was used to distinguish the producing areas according to the quantification results.Results 1.literature research:① the name of Tripterygium Wilfordii in ancient books is various,mainly including "Duanchang grass","yellow vine","Hu man";②Tripterygium wilfordii is a kind of plant likes wet,which is mostly grown in the hillside,valley and bush along the stream,and it is distributed in the South and southwest of the Yangtze River Basin in China;③ Tripterygium wilfordii is spicy,bitter and toxic,and it is mainly used for the treatment of muscle and bone pain and rheumatism arthralgia;and it is also used to prevent pest harm;④In response to Tripterygium wilfordii poisoning,the ancient times mainly infused animal blood,and the effect of amniotic blood was better.⑤.The disease distribution of tripterygium wilfordii treatment was at least as follows:rheumatoid arthritis 49.12%,nephritis 21.05%,diabetic nephropathy 9.65%,liver cancer 4.39%,systemic lupus erythematosus 3.51%,lung cancer 3.51%,leukemia 2.63%,Sjogren’s syndrome 2.63%,breast cancer 1.75%,intestinal cancer 1.75%.⑥The adverse reactions of clinical application of Tripterygum wilfordii were 24.32%of digestive system damage,15.94%of reproductive system damage,14.59%of bone marrow and blood system damage,13.78%of liver damage,11.62%of kidney damage,5.40%of skin damage,3.51%of cardiovascular system damage and 2.70%of poisoning death.⑦The research frequency of active components of tripterygium wilfordii from high to low was triptolide,triptolide alcohol,triptolide,triptolide ketone,triptolide A,triptolide,triptolide clonolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide,triptolide.44.63%,30.99%,22.52%,13.64%,12.45%,8.48%,7.68%,0.79%,0.26%,0.13%,respectively.2.The experimental research:The optimized parameters in QuEChERS method were list as follows:the extraction solvent was THF,buffer pH=1,4 g NaSO4 and lg CH3COONa,extraction time was 10 min,no adsorbents were used.Comparing with traditional heat-reflux method which takes 120 min extraction time,the developed QuEChERS method takes only 10 min to achieve 1.7 times higher extraction efficiency.Four procyanidins,Procyanidins B1;Catechin;Procyanidins B2;Epicatechin were identified by LC-MS together with comparison to reference standards.The antioxidant activity of crude extraction is 26.7 μg/mL,procyanidins B1 is 9.2 μg/mL,catechin is 7.1μg/mL,procyanidins B2 is 8.1 μg/mL,epicatechin is 7.6 μg/mL.These four components show better antioxidant activity compared with positive control BHT.3.The experimental researchThe content of these four components in seven different producing areas were quantified.The content differs from areas,some components have big differences in different areas.The static tool can group seven producing areas into four groups.Conclusion Tripterygium wilfordii have anti-inflammatory,antitumor,immunosuppression,and so on,mainly used for clinical treatment of rheumatoid arthritis,nephropathy,new autoimmune diseases such as systemic lupus erythematosus(sle),clinical adverse reaction is more,give priority to with the digestive and reproductive systems and liver and kidney damage with different degree,therefore to improve effect and decrease toxicity to research is of great significance.A novel QuEChERS method has been first developed in this paper.This method has been applied into extraction of four antioxidants in TWHF which shows higher extraction efficiency compared with traditional method.This method may be a potential standard flavonoids extraction method for natural products.Combing with LC-MS the major antioxidants can be identified and antioxidant activity can be screened.This strategy may become a new strategy for fast screening the antioxidants in natural products.Considering the production scale and contamination in mass spectrometer,ultrasound-assisted extraction method was developed to extract the antioxidants and followed by quantification of components by LC-MS.Different producing areas can be distinguished by principle component analysis.This may provide evidence for quality control of TWHF in different producing areas. |
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