Preparation method of 5-hydroxytryptamine derivatives from safflower seed meal by high-speed countercurrent chromatography
A technology of high-speed countercurrent chromatography and safflower seed meal is applied in the field of separation of natural medicines and achieves the effects of good repeatability, simple process and convenient operation.
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Embodiment 1
[0033] 1. Preparation of Crude Extract of 5-Hydroxytryptamine Derivatives
[0034] Take 300g of safflower seed meal and reflux extraction with absolute ethanol (liquid-to-material ratio: 15:1) for 1h, extract 3 times to combine the extract, evaporate to dryness with a rotary evaporator, dissolve in 500ml of 80% methanol, extract with an equal volume of n-hexane, and take the methanol phase Evaporate to dryness, dissolve in 300ml of ethyl acetate, then extract with an equal volume of water, and take the ethyl acetate phase and evaporate to dryness for later use.
[0035] 2. Preparation of 5-hydroxytryptamine derivatives by high-speed countercurrent separation
[0036] Prepare a two-phase solvent system of chloroform-methanol-0.1mol / L HCL water (1:1:1, v / v) in a separatory funnel, shake well and let stand overnight. The lower phase was the stationary phase, and the upper phase was the mobile phase, which were ultrasonically degassed for 30 min before use. Take 100 mg of the sa...
Embodiment 2
[0043] 1. Preparation of Crude Extract of 5-Hydroxytryptamine Derivatives
[0044] The preparation of the crude extract of 5-hydroxytryptamine derivatives is the same as in Example 1.
[0045] 2. Preparation of 5-hydroxytryptamine derivatives by high-speed countercurrent separation
[0046] Prepare a chloroform-methanol-0.1mol / L HCL water (3.5:4:4.5, v / v) two-phase solvent system in a separatory funnel, shake it well and let it stand overnight. The lower phase was the stationary phase, and the upper phase was the mobile phase, which were ultrasonically degassed for 30 min before use. Take 120mg of the sample powder to be separated, shake it and dissolve it completely in 10mL of the lower phase solution, and set it aside.
[0047] The lower phase of the solvent system was pumped into the main unit with a flow rate of 15 mL / min to fill the separation solenoid, and the circulating water bath was turned on and the temperature was set to 25 °C. Turn on the power of the main engi...
Embodiment 3
[0049] 1. Preparation of Crude Extract of 5-Hydroxytryptamine Derivatives
[0050] The preparation of the crude extract of 5-hydroxytryptamine derivatives is the same as in Example 1.
[0051] 2. Preparation of 5-hydroxytryptamine derivatives by high-speed countercurrent separation
[0052] Prepare a two-phase solvent system of chloroform-methanol-0.1mol / L HCL water (1:1:1, v / v) in a separatory funnel, shake well and let stand overnight. The lower phase was the stationary phase, and the upper phase was the mobile phase, which were ultrasonically degassed for 30 min before use. Take 100 mg of the sample powder to be separated, shake it and dissolve it completely in 10 mL of the lower phase solution, and set it aside.
[0053] The lower phase of the solvent system was pumped into the main unit with a flow rate of 15 mL / min to fill the separation solenoid, and the circulating water bath was turned on and the temperature was set to 25 °C. Turn on the power of the main engine an...
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