Methyl eicosapentaenoic acid preparing and purifying method from pavlova viridis
A technology of methyl eicosapentaenoate and Pavlova green algae, applied in chemical instruments and methods, preparation of carboxylate, preparation of organic compounds, etc., can solve the problems of low EPA yield and purity, and high cost, Achieve the effect of low cost, low consumption and high purity
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Embodiment 1
[0058] Example 1. Algae Cultivation and Harvesting
[0059] The unicellular marine microalga Pavlova viridis was purchased from the Institute of Oceanology, Chinese Academy of Sciences, Qingdao, rejuvenated and cultured in our laboratory, and stored at 21-27°C. The culture conditions are as follows: 6000-9000lux, 12:12L / D, aeration culture. Harvest at the end of exponential growth, centrifuge at 4000-8000g for 8-15 minutes to collect algae, wash with distilled water to remove salt and other impurities, freeze-dry, and store in low temperature freezer.
Embodiment 2
[0060] Embodiment 2. Preparation of silver-coated silica gel column:
[0061] Add a certain amount of silica gel for chromatography to twice the volume of absolute ethanol, stir and mix to form a suspension. Dissolve silver nitrate in an appropriate amount of 70% ethanol, add the above chromatographic silica gel and mix well. The weight ratio of silver nitrate to silica gel is 1:10. Then the ethanol was evaporated, and the obtained silver-coated silica gel was placed in an oven, activated at a high temperature of 120°C (more than 12 hours), cooled, and stored in a dark place.
Embodiment 3
[0062] Example 3. Preparation and purification of eicosapentaenoic acid methyl ester from Pavlova viridis
[0063] Preparation and purification of eicosapentaenoic acid methyl ester from Pavlova viridis, it comprises the following steps:
[0064] Step 1. Put 1.0 g of green Pavlova freeze-dried algae powder into a screw-top test tube, add 20 ml of 5% chloroacetyl-methanol (ie 1:20 v / v), fill it with nitrogen and seal it, and store it at 80°C Reacted for 2 hours, cooled to room temperature,
[0065] Step 2. Add equal volumes of double-distilled water and n-hexane to the reaction mixture obtained in step 1, shake and extract, absorb the upper liquid, then add an equal amount of n-hexane to repeat the extraction until the supernatant is no longer yellow,
[0066] Step 3. After combining the extracts, dry with a desiccant, filter the desiccant, and evaporate n-hexane under the protection of nitrogen to obtain 0.046 grams of fatty acid methyl esters,
[0067] Step 4. the fatty aci...
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