Preparation method for separating and purifying EPA and DHA
A separation, purification and semi-preparation technology, applied in the field of separation and purification of EPA and DHA, can solve the problems of high cost, complicated operation, low purity, etc., and achieve the effect of low production cost and simple post-treatment
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Embodiment 1
[0010] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 5mg / ml, put it in a filter device, and filter to remove solid particles;
[0011] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ10×250mm, C18 packing, the particle size is 5~45um, the sample volume is 10mg, CO 2 The flow rate is 0.8~3.0ml / min, methanol is used as modifier, the flow rate is 0~0.5ml / min, the temperature is 40°C, and the back pressure is 9~15MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-27min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.8% by HPLC analysis.
Embodiment 2
[0013] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 10mg / ml, put it in a filter device, and filter to remove solid particles;
[0014] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ10×250mm, C18 packing, the particle size is 5~45um, the sample volume is 20mg, CO 2 The flow rate is 1.0~5.0ml / min, methanol is used as modifier, the flow rate is 0~1.0ml / min, the temperature is 40°C, and the back pressure is 9~15MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-25min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.5% by HPLC analysis.
Embodiment 3
[0016] 1. Take the crude fish oil extract containing a mixture of EPA and DHA, dissolve it in methanol, the sample solubility is 10mg / ml, put it in a filter device, and filter to remove solid particles;
[0017] 2. Inject the sample solution into the semi-preparative SFC separation system, the chromatographic column size is Φ25×250mm, C18 filler, the particle size is 5~45um, the sample volume is 50mg, CO 2 The flow rate is 1.0~10.0ml / min, methanol is used as modifier, the flow rate is 0~2.0ml / min, the temperature is 40°C, and the back pressure is 10~18MPa. The detection wavelength of the ultraviolet photometric detector used is 210nm, and the fractions with retention times of 5-25min (corresponding to EPA) and 6-30min (corresponding to DHA) are collected, and the purity is ≥99.7% by HPLC analysis.
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