Researches On Chemicals Extraction Separation And The Antioxidant Active Material Basis In Paeonia Lactiflora Pall.'zhongjiang'

Paeonia lactiflora Pall.is a perennial herb in Paeonia L.genus Paeoniaceae family.It is not only a famous ornamental flower,but also a common medicinal material in China.P.lactiflora is often used to nourish the blood and regulate the menstrual,astringe yin and suppress sweating,soften liver and relieve pain,smooth liver-yang.Modern pharmacological research shows that P.lactiflora performs outstanding acts of anti-oxidation,antispasmodic,analgesic,anti-inflammatory,liver-protecting and immune-regulating.As a genuine traditional Chinese medicine,P.lactiflora‘Zhongjiang'is well-known in China and abroad,as its excellent quality.In this work,the HPLC fingerprints,extraction process,chemical composition and antioxidant material basis of P.lactiflora‘Zhongjiang'were studied.It aims to provide scientific theoretical basis for resource development and green sustainable development of P.lactiflora‘Zhongjiang'.The results of our achieved are as follows:1.The HPLC chromatographic fingerprint method of P.lactiflora‘Zhongjiang'was established.The chromatographic conditions were:Agilent Eclipse XDB-C18 column(4.6×250 mm,5?m),detection wavelength of 230 nm,column temperature of 40°C,injection volume of 10?L,flow rate 1.0 m L/min.The mobile phase consisted of water containing 0.1%H₃PO₄(solvent A)and acetonitrile(solvent B).The method applied in the following gradient elution orders were:10%?15%B for 0?5 min,15%?20%B for 5?20 min,20%?60%B for 20?40 min,60%?90%B for 40?45 min,90%B for45?50 min,90?10%B for 50?55 min,10%B for 55?60 min.A total of 20 common peaks were obtained,accounting for 94.6%of the total peak area.2.The TPA of HPLC fingerprint was used as the evaluation index to optimize the extraction conditions of ultrasound-assisted extraction(UAE),microwave assisted extraction(MAE),and subcritical water extraction(Sub WE).The optimal process conditions of UAE were:72%ethanol,ultrasonic power of 360 W,ultrasonic time of 49min,liquid-to-solid ratio of 19 m L/g.Under these conditions,TPA was(7.21±0.13)×10⁸.The optimal process conditions of MAE were:65%ethanol,extraction temperature of62°C,liquid-to-solid ratio of 19 m L/g,extraction time of 11 min.Under these conditions,TPA is(6.69±0.19)×10⁸.The optimal process conditions of Sub WE were:subcritical water temperature of 184±5°C,liquid-to-solid ratio of 17 m L/g,extraction time of 18.0min.Under these conditions,TPA was(2.81±0.12)×10⁸,and the extraction rate of paeoniflorin was 3.26%.By comparison of the three methods,the suitable technics for the subsequent preparation of medical components and research on the antioxidant active material basis of P.lactiflora‘Zhongjiang'.respectively were obtained.3.For the first time established the Sub WE technology conditions of total glucosides of P.lactiflora‘Zhongjiang':extraction time of 22 min,extraction temperature of 158°C,and liquid-to-solid ratio of 21 m L/g.The total glycoside extraction rate was 7.08±0.20%under the optimal condition.After treatment with AB–8 resin,the content of TGP increased significantly in the purified product from 32.5%to 89.6%,and the content of paeoniflorin and albiflorin in total glucosides of paeoniflorin are 63%and 25%respectively.The optimal MAE technology conditions of paeoniflorin from the stems and leaves of P.lactiflora were established:70%Et OH,extraction temperature of 61°C,extraction time of 9.5 min,and the liquid-to-solid ratio of 22 m L/g.The paeoniflorin extraction rate was 2.81±0.19%under the optimal condition.The content of paeoniflorin was>95%by purification.4.From the root of P.lactiflora‘Zhongjiang',28 compounds were isolated and structurally elucidated.These include 12 monoterpenoid glycosides,7 phenolic acids and their ester compounds,1 flavonoid,and 8 other compounds.The compounds benzoylpaeoniflorin(11),albiflorin(19),paeoniflorin(20),and gallic acid(21)were the main components.These studies have improved the chemical composition data and provided the basis for the research of the antioxidant active material basis of P.lactiflora‘Zhongjiang'.5.A total of 40 compounds,including 19 monoterpene glycosides,5 tannics,10phenolic acids and their esters,and 6 other compounds,were identified or tentatively deduced by HPLC-DAD-ESI-MS.35 compounds were found in roots,20 compounds in stems and leaves,15 compounds in flowers.6.The antioxidant activities of extracts from roots,stems with leaves,and flowers,as well as some monomeric compounds were determined by DPPH and ABTS spectrophotometry.The results showed that the extracts are of strong antioxidant activity;Catechin,gallic acid and ethyl gallate have strong antioxidant activity,and the extent of antioxidant activity is notable concentration dependent.Paeoniflorin,albiflorin,benzoylpaeoniflorin and 6'-O-benzoylalbiflorin are in low some antioxidant activity,but low activity.19,15 and 15 potential antioxidants were found from roots,stems and leaves,and flowers extracts by DPPH-HPLC experiment,respectively.The results showed that the ethanol extracts of the roots,stems and leaves,and flowers of P.lactiflora‘Zhongjiang'have strong antioxidant activity.The material foundation of antioxidant activity of P.lactiflora‘Zhongjiang'is mainly phenolic acids and tannins compounds.The monoterpene glycosides,the main component of the P.lactiflora,have only weak antioxidant capacity.