Method for purifying EPA (eicosapentaenoic acid) from fish oil
A technology of EPA-C18 and fish oil, applied in the separation/purification of carboxylic acid compounds, organic chemistry, etc., can solve the problems of inability to separate EPA and DHA, ineffective separation of EPA and DHA, slow hydrolysis speed, etc., and it is not easy to achieve Oxidation, easy operation, high recovery effect
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Embodiment 1
[0100] Example 1 Fish oil crude product analysis before purification
[0101] The crude fish oil has a purity of 70%.
[0102]Chromatographic column: Sepax GP-C18 (5um, 4.6x 250mm)
[0103] mobile phase:
[0104] A: water
[0105] B: Acetonitrile
[0106] Flow rate: 1ml / min Injection volume: 5ul Column temperature: RT
[0107] Sample: 1mg / ml (dissolved in 82% methanol) Pressure: 78bar
[0108] Instrument: Agilent 1260
[0109] Detection wavelength: UV@214nm
[0110] Elution was performed according to the following gradient:
[0111] Table 2
[0112] Time min)
[0113] The crude product analysis diagram is as follows figure 2 shown.
Embodiment 2
[0114] Embodiment 2 fish oil crude product purification
[0115] The crude fish oil has a purity of 20%.
[0116] Chromatographic column: EPA-C18 (1000x 250mm)
[0117] mobile phase:
[0118] A: water
[0119] B: Methanol
[0120] Flow rate: 400ml / min Injection volume: 400ml Column temperature: RT
[0121] Sample: 100mg / ml Pressure: 1.2MPa
[0122] Instrument: Sepax preparative chromatograph
[0123] Detection wavelength: UV@214nm
[0124] Gradient: 82%B
[0125] The specific method is as follows:
[0126] (1) Sample weighing: weigh 40 g of fish oil crude product on an analytical balance;
[0127] (2) Solvent preparation: prepare 400ml of 82% methanol, measure 328ml of methanol, and measure 72ml of purified water, mix the two together, and stir evenly, prepare two parts, and set aside;
[0128] (3) Sample dissolution: add one part of the solvent prepared in (2) above to (1) 40g of crude fish oil, seal it with plastic wrap, add a stir bar, and keep stirring on a magne...
Embodiment 3
[0136] Get fish oil crude product 40g, wherein EPA content 70% (m / m), add 82% methyl alcohol (V / V) 400ml, stir and make fully dissolving; Upper chromatographic column EPA-C18 (100mm * 250mm , containing packing quality about 1.18kg), sample loading flow rate 200ml / min, elution with 82% methanol after sample loading, elution flow rate 400ml / min, elution total 100min, HPLC detection and collection of EPA component solution, membrane concentration, used Nanofiltration membrane area 2.54m 2 , molecular weight cut off 150~300, membrane flux 20L / (m 2 · h), the temperature is controlled at 0-50° C., and the concentrated component solution is dried to obtain 26.2 g of EPA product. The purity detected by HPLC is 97.3%, and the recovery rate is 91.1%.
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