A method for separating phycoerythrin in Pteridophyllum pinnata using high-speed countercurrent chromatography
A technology for erythropoietin and high-speed counter-current chromatography in the pinnate algae, which is applied in the direction of organic chemistry, etc., can solve the problems of not many methods for separation and purification of erythropoietin from fiddlehead algae and less algae in pinnate algae, and achieves good application value, The effect of large amount of preparation and elimination of errors
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Embodiment 1
[0048] (1) Accurately weigh 20.00g of dry Pteridophyllum, use microwave-assisted extraction method to extract the phycoerythrin in Pteridophyllum, obtain the crude extract, the extraction conditions are as follows: extraction solvent 90% (volume percentage) ethanol , microwave temperature 50°C, microwave power 300W, microwave time 15min, solid-liquid ratio 1:40g / mL.
[0049] Table 1 Factor and level design of single factor experiment
[0050]
[0051] When the solid-liquid ratio is 1:40-1:50g / mL, the extraction rate does not increase significantly. Considering the extraction cost and the difficulty of subsequent purification, the solid-liquid ratio is 1:40g / mL. The microwave-assisted extraction parameters of this embodiment were then determined through the orthogonal experiment in Table 2 below.
[0052] Table 2 Orthogonal test factors and levels
[0053]
[0054] Finally, the microwave temperature is 50°C, the microwave time is 15min, the microwave power is 300W, the ...
Embodiment 2
[0066] (1) Accurately weigh 20.00g of Pteris pinnata, after drying, use microwave-assisted extraction method to extract phycoerythrin in Pteris pinnata, the extraction conditions are as follows: extraction solvent 95% ethanol, microwave temperature 60°C, microwave power 100W , microwave time 5min, solid-liquid ratio 1:30g / mL.
[0067] (2) Select the solvent system for high-speed countercurrent chromatography: n-hexane-methanol-water (2:1:1, v:v:v).
[0068] The upper phase of the solvent system is used as the stationary phase, and the lower phase is used as the mobile phase. The retention rate of the stationary phase is about 76.8%.
[0069] (3) Concentrate the crude extract into an extract, dissolve it in the mobile phase of the HSCCC solvent system, and separate it. The separation conditions are: flow rate 1.0mL / min, rotation speed 800rpm / min, detection wavelength 315nm.
[0070] (4) The separation fraction obtained and the pteridin standard substance were injected into hi...
Embodiment 3
[0073] (1) Accurately weigh 20.00 g of Pteridophyllum pinnatis, and extract the phycoerythrin in Pteridophyllum pinnata by microwave-assisted extraction. The extraction conditions are as follows: extraction solvent 85% ethanol, microwave temperature 40°C, microwave power 200W, microwave time 10min, solid-liquid ratio 1:50g / mL.
[0074] (2) Select the solvent system for high-speed countercurrent chromatography: n-hexane-methanol-water (2:1:1, v:v:v).
[0075] The upper phase of the solvent system is used as the stationary phase, and the lower phase is used as the mobile phase. The retention rate of the stationary phase is about 76.8%.
[0076] (3) Concentrate the crude extract into an extract, dissolve it in the mobile phase of HSCCC solvent system, and separate it. The separation conditions are: flow rate 3.0mL / min, rotation speed 900rpm / min, detection wavelength 315nm.
[0077] The obtained separated fraction and the phycoerythrin standard sample were injected into high-per...
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