Extraction,Isolation And Biological Of Chemical Components From Pterocaryarhoifolia

Pterocarya plants are widely distributed in our country,It is mainly distributed in North China,central China,East China,South China and Southwest China.It is mostly found near streams or on wet hillsides.Its branches,leaves,flowers and fruits not only have high ornamental value,but also have rich pharmacological activities and harmful biological control functions,such as tumor inhibitory activity,antioxidant,antibacterial,anti-inflammatory,treatment of fish disease and snails,etc.In folk,this genus of plants is a commonly used Chinese medicine,can treat a variety of bacterial and fungal skin diseases.However,research on the genus Pterocarya is limited to a few species such as Pterocaryatonkinesis and Pterocaryahupehensis,and no systematic studies have been conducted on the composition of its active ingredients,related pharmacological activities and their uses,and almost no research has been conducted on Pterocaryarhoifolia,which is endemic to Shandong.In order to better understand the chemical constituents of this genus,this paper systematically isolated and purified the chemical constituents from the leaves of this genus Pterocaryarhoifolia,and preliminarily studied the pharmacological activities of this genus.The main research contents and achievements include:(1)A preliminary investigation of the activity of crude extracts of Pterocaryarhoifolia leaves.The extracts were obtained from Pterocaryarhoifolialeaves by soaking in 85%ethanol as the extraction solvent for a certain period of time,followed by extraction with petroleum ether,ethyl acetate and n-butanol to obtain the different polar parts.The preliminary determination of the antioxidant,anti-inflammatory,antibacterial and anti-tumour biological activities of each part showed that the ethyl acetate part was the most active.(2)Systematic isolation and purification of the most active ethyl acetate site.The ethyl acetate extracted phase of the ethanolic extract of Pterocaryarhoifolia leaves was purified by silica gel column and gel column chromatography(Sephadex LH-20)for several times,and its structure was identified and physicochemical properties were analyzed.Finally,The four compounds obtained were(4S)-4,8-dihydroxy-1-tetralone(56mg),a naphthoquinone,4-methoxy-5-hydroxy-1-tetralone(17 mg),the flavonoid myricetin(22 mg)and β-sitosterol(50 mg),a steroid-like compound.The antitumor effects of(4S)-4,8-dihydroxy-1-tetralone and myricetin were determined by MTT method,The results showed that(4S)-4,8-dihydroxy-1-tetralone and myricetin showed antagonistic effects on the proliferation of human hepatocellular carcinoma cells Hep-G2 and neuroblastoma cells SH-SY5 Y,the IC50 values were 74.99±2.14 μg/mL,54.50±1.97μg/mL,63.99±2.67 μg/mL,37.13±2.31 μg/mL。(3)Determination of the composition of the volatile components of the crude petroleum ether extract of Pterocaryarhoifolia leaves by gas chromatography coupled with mass spectrometry(GC-MS).The GM-MS results showed that most of the components in the crude extract of petroleum ether from Pterocaryarhoifolia leaves were dominated by fatty acids,such as n-Hexadecanoic acid with a relative content of 36.92%,(Z,Z,Z)-9,12,15-Octadecatrienoic acid with 26.50%,(Z,Z)-9,12-Octadecadienoic acid with12.21%,Tetradecanoic acid with 5.54%and Octadecanoic acid with1.57%.In addition to these substances,there are other substances such as alkanes,olefins and phytols.(4)The optimal extraction process of naphthoquinones and flavonoids from Pterocaryarhoifolia leaves was investigated using response surface methodology and orthogonal tests.The main factors affecting the extraction content of naphthoquinones and flavonoids were firstly clarified by single-factor experiments,and then the extraction processes of naphthoquinones and flavonoids were optimized by response surface methodology and orthogonal tests,respectively.The results showed that the extraction of naphthoquinones was optimal when 60% ethanol,1:40(g/mL)material-to-liquid ratio,50 ℃ extraction temperature and 6 h extraction time were used as the extraction conditions for naphthoquinones,and the extraction amount was 106.37 mg/g;The extraction effect was optimal when 75% ethanol,1:30(g/mL)material-to-liquid ratio,180 W sonication power and 45 min sonication time were used as the extraction conditions for the flavonoids,and the extraction amount was 28.54 mg/g.