Determination Of Compatibility Safflower Seed Oil By Gas Chromatography And Study On Its Effect Of Anti-cells-damage Mediated By Oxygen Free Radical

Objective: To apply the GC external reference method determining the contents of linoleic acid (LA) andα-linolenic acid (α-ALA) in different proportion Compatibility Safflower Seed Oil (CSSO), and to establish the assaying method of CSSO; To establish SH-SY5Y cell oxygen damage model induced by H₂O₂; To investigate the effects of different proportion compatibility safflower seed oil at oxidative damage of SH-SY5Y cells mediated by oxygen free radical, find the best compatibility proportion of CSSO in prophylaxis SH-SY5Y cells oxygen damage and the effective concentration range. Methods:Prepare different proportion compatibility safflower seed oil. Choose methyl linoleate and methylα-linolenate as the external reference substance ofα-linolenic acid and linoleic acid and determine the contents of them in CSSO; Establish oxygen damage cell model through the experiments of H₂O₂ concentration and time gradient and choose the concentration of 0.7% DMSO as solvent to set up different proportion compatibility safflower seed oil gradient in medium culture with SH-SY5Y cells for 24h.Cell morphology was observed by microscopy, cell viability and cellular apoptosis were tested by a colorimetric assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and flow cytometry, respectively. With statistical analysis to analyze the effects of different proportion CSSO in prophylaxis cells oxygen damage. Results:The GC contents determination assay of CSSO was established, definition the chromatographic conditions, determined the relative contents of LA and ALA in safflower oil, mix-PUFA and three proportions CSSO. After 3 hours induced by 1000μM H₂O₂, about half SH-SY5Y cells morphous become abnormal, cell viability decreased significantly(P<0.01) and the inhibition ratio is 46.8%. Different proportion of CSSO all have prophylaxis effect on SH-SY5Y cells oxygen damage induced by H₂O₂ in different extent after 24h.The ratio of ALA and LA at 1:6.68 has the best effectiveness. MTT results show that at the concentration range of 375μg/ml-750μg/ml cell viability is increased gradually with the concentration, at the concentration of 750μg/ml has the highest cell viability of 84.14%. This result has an in concordance with the result of cellular apoptosis tested by flow cytometry. Conclusions: GC determination method of CSSO has satisfactory repetitiveness, precision and recovery rate and the chromatographic condition meet the request of contents determination of CSSO. Set up a relatively stable and repeatability SH-SY5Y cell oxygen damage model induced by H₂O₂. The proportion of LA and ALA at 1:6.68 has a relative better effect in prophylaxis oxygen damage, which shows it has potential use value in clinic as a prophylaxis ND component.