Study On Purification And Its Antioxidation And Antitumor Activity Of Pinolenic Acid In Pine Nut Oil

In this study,Pinus massoniana was used as raw material to extract pine nut oil,and pinolenic acid(PLA)in pine nut oil was isolated and purified.The antioxidant and antitumor activities of PLA were also studied in vitro.The results showed as follows:(1)Pine nut oil and pine seed meal were extracted by centrifugal extraction.The heating temperature was 85?,centrifugal speed was 6500rpm(Centrifuged at 4487 × g)and centrifugation time was 15min.The extraction rate of pine nut oil was 60.62%;(2)Urea inclusion was used to extract PLA,and the purity of PLA was analyzed by GC-MS.The results showed that the ratio of urea to fatty acid was 3.5:1,urea:ethanol was 1:5,inclusion time was 24 h,crystallization temperature at 0?.In that condition,the yield of PLA was 22.19%,and the purity of PLA was increased from 20.61%to 72.45%;(3)The method of silver nitrate silica gel column chromatography was carried out on the basis of the urea inclusion method.The ratio of the amount of sample to the adsorbent was 1:20 and the flow rate was 1.5-2.OmL/min.Under these conditions,PLA was eluted at the concentration of 4%acetone(acetone accounted for the total volume of the mass fraction),and flowed out at 8%concentration separation completely,after column chromatography,the purity and yield of PLA was 99%and 44.32%,respectively;(4)The functional groups of purified PLA were characterized by FT-IR.The results showed that the purified product had double bond,carboxyl group,-CH3 and-CH2-functional information.The 'H NMR and 13C NMR spectra con firmed that Molecular Structure:CH3(CH2)4CH=CH-CH2-CH=CH(CH2)2CH= CH(CH2)3COOH;(5)The antioxidant activity of PLA in vitro showed that PLA had almost no effect on DPPH·and·OH radical scavenging,but had obvious inhibitory effect on lipid peroxidation;(6)In vitro antitumor activity of PLA:MTT assay showed that PLA could inhibit the proliferation of Hela cells in a time and dose-dependent manner,and the IC50 of Hela cells at 48h and 72h were 45.48?g/mL?38.308?g/mL,respectively.The results of scratches showed that PLA could inhibit the migration of Hela after 48h.The apoptosis of Hela cells was observed by HE staining.The cell apoptosis was observed by flow cytometry,the cells were repressed in G0/G1 and S phase,which indicated that PLA could induce Hela apoptosis.