Extraction and preparation method for limonin components
A technology of limonin and extract, which is applied in the field of biomedicine and can solve problems that have not yet been discovered
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experiment example 1
[0029] Experimental example 1: Determination of total citrin content.
[0030] 1. Preparation of reagents
[0031] (1) Limonin standard solution: Accurately weigh 0.0092g of limonin with an electronic balance, dissolve it with an appropriate amount of distilled water, transfer it to a 50mL volumetric flask after dissolving, constant volume, and shake well to obtain a mass concentration of 184mg / L Limonin standard solution;
[0032] (2) Display agent: Measure 65mL of concentrated sulfuric acid and 35mL of absolute ethanol, mix them, and add 125mg of p-dimethylaminobenzaldehyde after cooling. When in use, add 0.05mL of 9% ferric chloride solution to the solution and mix it evenly, and prepare it now;
[0033] (3) 9% ferric chloride solution: Weigh 4.5 g of ferric chloride solid, dissolve the solid in 45.5 mL of distilled water, and transfer it to a brown reagent bottle for storage after cooling.
[0034] 2. Make a standard curve
[0035] Take 6 test tubes, add 0, 0.4, 0.8,...
experiment example 2
[0038] Experimental example 2: HPLC-MS qualitative analysis of the total limonin extract of Ragbu japonica.
[0039] Preparation of the sample solution: take the total limonin extract of the ragwort fruit, prepare the test solution with methanol, filter it with a 0.45um organic microporous membrane, and use it as the test solution.
[0040]Preparation of standard solution: take homemade limonin, nomilin, limonoid diomol, leucodiol, 1-deacetylniporin B, cortex ketone, Yichangsu, granatumin H, swimacronoid A , add an appropriate amount of methanol as a standard solution.
[0041] Instrument: German Thermo Fischer Scientific high-resolution liquid chromatography-mass spectrometer.
[0042] Chromatographic conditions: Waters Symmetry C18 column (5μm, 250mm × 4.6mm), column temperature: 25°C, flow rate 800μl / min, detection wavelength: 550nm, injection volume: 5ul. Mobile phase: use acetonitrile as mobility A, 0.1% ammonia water as mobility B, carry out gradient elution as specifi...
Embodiment 1
[0047] Example 1: Preparation of total limonin extract and monomeric compound of Ragbukia
[0048] (1) 15kg of Ragwood fruit, use 80% ethanol as solvent, extraction temperature is minus 7°C, extraction times are 3 times, each extraction time is 7 hours, each time the amount of solvent is 15 times of the weight of Ragwood fruit, Filtrate, and freeze-dry the extract to obtain the crude extract of Ragbukia fruit;
[0049] (2) Purify the crude extract of Ragbukia fruit obtained in step (1) through MCI gel column chromatography, first elute with methanol-water with a part ratio of 2:98, and then use a part ratio of 10:90 Methanol-water elution, collect the methanol-water elution part with a fraction ratio of 10:90, recover methanol, and dry to obtain the ragwort fruit extract;
[0050] (3) Put the ragwort fruit extract obtained in step (2) through an MCI gel-silica gel mixed chromatographic column with a weight ratio of 1:1, and use petroleum ether-acetone gradient elution, and us...
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