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Method for preparing flavonoids compounds in camellia seed shells by high-speed counter-current chromatography

A technology of high-speed countercurrent chromatography and flavonoids, applied in chemical instruments and methods, preparation of sugar derivatives, sugar derivatives, etc.

Inactive Publication Date: 2015-08-26
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] High-speed countercurrent chromatography is a new type of continuous and efficient liquid-liquid partition chromatography technology, which has been widely used in the fields of biology, medicine, food, materials, etc., especially in the field of separation and purification of active ingredients of natural products. However, the method for separating and preparing flavonoids from camellia oleifera seed shells with this high-speed countercurrent chromatography has not been reported yet.

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  • Method for preparing flavonoids compounds in camellia seed shells by high-speed counter-current chromatography
  • Method for preparing flavonoids compounds in camellia seed shells by high-speed counter-current chromatography
  • Method for preparing flavonoids compounds in camellia seed shells by high-speed counter-current chromatography

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Preparation of flavonoids in camellia oleifera seed shell

[0018] Camellia oleifera seed shells in this example were purchased from Hongyuan Agriculture and Forestry Development Co., Ltd., Huaihua City, Hunan Province.

[0019] 1. Preparation of raw material (camellia oleifera seed husk extract) for high-speed countercurrent (HSCCC) loading

[0020] Take 200 g of Camellia oleifera seed shell powder that has been crushed through a 10-mesh sieve, extract three times at 60°C with 70% ethanol of 6 times the weight of Camellia oleifera seed shell powder, the first time is 60 minutes, and the last two times are 30 minutes each, and the extract is collected , combined, filtered, concentrated under reduced pressure at 55°C to recover ethanol, and the obtained water layer (extract) was adsorbed by a chromatographic column equipped with 300ml D-101 macroporous adsorption resin at a flow rate of 3BV / h, and left to stand for 30min after adsorption , first rinse with 900...

Embodiment 2

[0033] see in conjunction image 3 and Figure 4 According to the chromatogram of HPLC (analytical conditions as above) at 254nm, the purity of the compound was calculated by the peak area normalization method, the purity of compound 1 was 99%, and the purity of compound 2 was 93%. The structural identification of embodiment 2 compound

[0034] The structures of the above two compounds were identified by mass spectrometry and nuclear magnetic resonance.

[0035] Compound 1: light yellow powder, molecular weight 756.21, component formula C 33 h 40 o 20 ,according to 1 H and 13 C NMR data, combined with the literature, identified the compound as: kaempferol-3-O-[β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranose-(1→6 )-β-D-glucopyranoside], its structure is as follows, and the NMR signal assignment is shown in Table 2.

[0036]

[0037] Table 2 The H NMR and C NMR spectra of compound 1

[0038]

[0039]

[0040] Compound 2: light yellow powder, molecular weight 730.20...

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Abstract

A method for preparing flavonoids compounds in camellia seed shells by high-speed counter-current chromatography comprises the following steps: camellia seed shells are taken as raw materials which are crushed and then extracted with ethanol, the extracting solution is collected and purified by D-101 macroporous resin, and after that, the materials of water, n-butyl alcohol, methyl tert-butyl ether, acetonitrile and glacial acetic acid at the volume ratio of 6: 3: 1:1: 0.001 serve as a high-speed counter-current solvent system to conduct high-speed counter-current chromatography on the camellia seed shell extract to prepare two flavonoids compounds. The method is simple and convenient, and excellent in reproducibility, can be used for large-scale preparation of compounds and lays a material basis for further study on activity.

Description

technical field [0001] The invention relates to a method for preparing flavonoids, in particular to a method for separating and preparing flavonoids from camellia oleifera seed shells by high-speed countercurrent chromatography. Background technique [0002] Camellia oleifera (Camellia oleifera) belongs to the genus Camellia oleifera. The seeds can be squeezed for oil and eaten. The tea seed shell is the outer testa. The testa contains more pigments and is brown-black and hard. It accounts for 30.6%-34.0% of the weight of the tea fruit. %. Camellia oleifera is one of the main woody oil tree species in my country, with an annual output of 1.5 million tons of oil. Tea seed shell is a by-product of tea seed oil extraction, which is the seed coat removed before oil extraction, but its utilization rate is relatively low. Low. Studies have shown that tea seed husks are rich in tea saponin, flavonoids, polysaccharides, polyphenols, proanthocyanidins and other components. If tea se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/07C07H1/08
CPCC07H1/08C07H17/07
Inventor 陆英李觅路李嘉伟刘毅
Owner HUNAN AGRICULTURAL UNIV
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