A method for extracting different forms of fat-soluble polyphenols in Camellia oleifera seed oil
A technology of camellia oleifera seed oil and extraction method, which is applied in the fields of fat oil/fat production, non-central analgesics, pharmaceutical formulations, etc., can solve problems such as co-extraction interference, achieve improved extraction efficiency, and high antioxidant and anti-inflammatory activities , rich effect
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Embodiment 1
[0034] (1) Add the extraction solvent of 225mL n-hexane-acetonitrile-70% aqueous methanol (mixed in the ratio of 1:1.5:1.5 by volume) to 15g Camellia oleifera seed oil, shake vigorously at 37 ℃, ultrasonic power is Ultrasonic extraction under the condition of 350W for 45min. Then centrifuge at 4° C. and 8000 rpm for 15 min, collect the lower supernatant, and repeat the extraction of the upper oil phase for 2-3 times under the same conditions. The lower layers were combined and collected, and the organic solvent was removed by rotary evaporation, and the obtained aqueous phase was the soluble polyphenol extract, which was used for the subsequent extraction of free, esterified and glycosylated polyphenols.
[0035] (2) Use 6M HCl to acidify the aqueous phase collected in step (1) to pH 3, and then use the same volume of diethyl ether-ethyl acetate mixed solvent (1:1, v / v) to Under the condition of 350W, ultrasonically extract the aqueous phase 4 times, 45min each time. Then ce...
Embodiment 2
[0044] The content of polyphenols refers to the literature report (Gouvinhas I, Machado J, Gomes S, et al: Phenolic composition and antioxidant activity of monovarietal and commercial portuguese olive oils. J Am Oil Chem Soc 2014, 91 (7): 1197-1203) using Forin ratio Color method, using gallic acid as a standard to determine the content of different forms of fat-soluble polyphenols in camellia oleifera seed oil. The standard solution concentration gradients are: 0, 20, 40, 60, 80, 100 μg / mL. Add 5mL standard solution or polyphenol sample diluted to an appropriate multiple, then add 0.5mL Folin phenol reagent, mix well and let stand for 2min. Then add 1 mL of 10% Na 2 CO 3 Aqueous solution and dilute to 10mL, mix and let stand for 1.5h. Absorbance was measured at a wavelength of 765 nm. The polyphenol content in the sample was calculated according to the standard curve, expressed as gallic acid equivalent (GAE).
[0045] The results showed that the contents of free, esteri...
Embodiment 3
[0047] Chemical antioxidation refers to literature reports (Xia Yu, Zhang Hui, Xiao Junyong, Cong Renhuai, Qian Haifeng, Wang Li, Qi Xiguang. Effect of oil production process on grape seed oil phenolics and its antioxidant activity. China Oils, 2018, 43 (06): 1-6.) with ORAC FL The chemical antioxidant activity of different forms of fat-soluble polyphenols in camellia oleifera seed oil was determined by using Trolox as a standard. Prepare 75mmol / L disodium hydrogen phosphate solution and potassium dihydrogen phosphate solution with a pH meter to prepare a buffer solution with a pH of 7.4. Prepare 8.16×10 with the above-mentioned phosphate buffer solution -5 mmol / L fluorescein sodium salt solution and 153mmol / L AAPH reaction solution. The standard solution concentration gradients are: 3, 6, 12, 24, 48, 96 μmol / L. In a dark state, use a 100 μL pipette gun to pipette the polyphenol extract or 25 μL Trolox standard working solution and add it to the 96-well black microtiter pla...
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