New method for extracting and separating high-purity huperzine A from thousand-layer column
A technology of huperzine A and Melaleuca tower, which is applied in the field of extraction and separation of high-purity huperzine A, can solve the problems of not meeting market demand, small production scale, and low product purity, and achieve saving of reagents, short process cycle, The effect of high purity
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[0027] Preparation of the extract: ① Add 3-25 times the amount of 0.2-2% sulfuric acid to the crushed Melaleuca tower and soak for 0-24 hours at room temperature; ② Percolate extraction until no Huperzine A can be detected in the effluent. The extract is reserved;
[0028] Macroporous resin pre-separation: ①Adjust the pH of the extract to 8-10, and apply to a macroporous adsorption resin column; ②Use 0-15% ethanol for the first elution, and discard the eluate; ③Use 15 for the second time -Elute with 70% ethanol, collect the eluate, and recover the ethanol to obtain the extract extract;
[0029] Purification by high-speed countercurrent chromatography: Using petroleum ether-n-butanol-phosphate buffer as a solvent system, the crude extract of Huperzine A obtained above is purified by high-speed countercurrent chromatography to obtain high-purity Huperzine A. The purity is above 98%.
Example Embodiment
[0030] Example 1:
[0031] Take 100g of the crushed Melaleuca medicinal material, add 1.5L of 1% sulfuric acid, soak at room temperature for 24 hours, percolate until no Huperzine A is detected in the effluent, adjust the pH of the extract to 8 with ammonia.
[0032] Put the pH-adjusted extract on a macroporous adsorption resin column, eluting with 5% ethanol until the effluent is colorless, and the eluate is discarded. Then it was eluted with 30% ethanol to detect the absence of Huperzine A in the eluate, collect the eluate, and distill under reduced pressure to recover the solvent to obtain an extract of Huperzine A.
[0033] Use petroleum ether-n-butanol-phosphate buffer as the high-speed countercurrent chromatography two-phase solvent system to separate and purify the extract of Huperzine A, the stationary phase is petroleum ether-n-butanol (1.5:3.5, V / V), mobile The phase is a phosphate buffer containing 1% sodium dihydrogen phosphate and disodium hydrogen phosphate.
Example Embodiment
[0034] Example 2:
[0035] Take 100g of the crushed Melaleuca medicinal materials, add 1.5L of 1% sulfuric acid, soak at room temperature for 24 hours, and ultrasonically treat for 30 minutes to separate the medicinal solution. 1L of 1% sulfuric acid was added to the medicine residue, and ultrasonic treatment was used for 20 minutes to separate the medicine liquid. Combine the medicinal solutions, filter to remove impurities in the extract, and adjust the pH of the extract to 8.
[0036] Put the pH-adjusted extract on a macroporous adsorption resin column, eluting with 5% ethanol until the effluent is colorless, and the eluate is discarded. Then eluted with 30% ethanol until there was no Huperzine A in the effluent, collected the eluate, and distilled under reduced pressure to recover the solvent to obtain an extract of Huperzine A.
[0037] Use petroleum ether-n-butanol-phosphate buffer as the high-speed countercurrent chromatography two-phase solvent system to separate and purify...
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