Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for preparing high-purity hydroxytyrosol by using high-speed counter-current chromatography and high performance liquid chromatography in combined manner

A technology of high-speed countercurrent chromatography and high-performance liquid chromatography, which is applied in chemical instruments and methods, preparation of organic compounds, organic chemistry, etc., can solve the problem of high-purity hydroxytyrosol without batches, achieve fewer steps, increase yield, high cost effect

Active Publication Date: 2015-12-16
INST OF CHEM IND OF FOREST PROD CHINESE ACAD OF FORESTRY
View PDF4 Cites 16 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the separation and preparation of hydroxytyrosol by high-speed countercurrent chromatography, and there is no batch of high-purity hydroxytyrosol products on the international market

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for preparing high-purity hydroxytyrosol by using high-speed counter-current chromatography and high performance liquid chromatography in combined manner
  • Method for preparing high-purity hydroxytyrosol by using high-speed counter-current chromatography and high performance liquid chromatography in combined manner
  • Method for preparing high-purity hydroxytyrosol by using high-speed counter-current chromatography and high performance liquid chromatography in combined manner

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Separation of standard samples by high-speed countercurrent chromatography

[0033] 1. Experimental equipment:

[0034] FastChrome analytical high-speed countercurrent chromatograph (Jiangyin Countercurrent Technology Co., Ltd.); separation column volume: 25ml; value range: 0.56~0.91; OptiChromeA semi-preparative high-speed countercurrent chromatograph (Jiangyin Countercurrent Technology Co., Ltd.); spiral tube separation column (revolution Radius: 80cm; Value range: 0.50~0.80; Spiral tube inner diameter: 1.59mm), single column volume 180ml, double column volume 360ml.

[0035] 2. Experimental materials:

[0036] Analysis of pure isomers of phenolic samples: catechol, resorcinol, hydroquinone; n-hexane, ethyl acetate, methanol, distilled water

[0037] 3. Preparation of solvents and samples:

[0038] Prepare the methanol mixed solution of the phenolic sample, the concentration of the mixed sample: catechol 20mg / ml+ resorcinol 20mg / ml+ hydroquinone 10mg / ml; use 80% metha...

Embodiment 2

[0059] Example 2 High-speed countercurrent chromatography and high-performance liquid chromatography combined method

[0060] The first step, preparation of hydroxytyrosol ointment

[0061] The oleuropein extract of Olea europaea is enzymatically hydrolyzed, the solid-liquid ratio of the extract and the phosphate buffer solution of 4-7 is 1g:10-100mL, and the mass ratio of the extract and the biological enzyme is 1g:20-100mg, placed in 30 ℃~100℃ in a constant temperature water bath shaker. After enzymolysis for 1h-10h, filter, the filtrate is extracted with ethyl acetate, the ethyl acetate layer is concentrated and dried, the concentrate is hydroxytyrosol ointment, the yield of the ointment relative to the plant leaf powder is 1-20%, and the hydroxytyrosol content is 5~ 20%;

[0062] The second step, high-speed countercurrent chromatography separation

[0063] 1) Select the 3-6:4-8:3-6:4-8 solution of petroleum ether: ethyl acetate: methanol: water according to the volume ratio as t...

Embodiment 3

[0068] Example 3 Experiment of separating hydroxytyrosol ointment by high-speed countercurrent chromatography

[0069] 1. Solvent preparation: petroleum ether: ethyl acetate: methanol: water=2:3:2:3. Prepare 2000ml according to the volume ratio. The upper phase is the stationary phase and the lower phase is the mobile phase.

[0070] 2. Sample preparation: Take 50 mg of hydroxytyrosol ointment in Example 3 and dissolve it in 50 ml of mobile phase.

[0071] 3. System balance: pump full stationary phase at a speed of 50ml / min, pump in mobile phase at a speed of 5ml / min, start

[0072] 4.OptiChrome TM A-300 forward centrifugation, observe the UV3000 output to reach the system balance, calculate the stationary phase retention rate: 60%

[0073] 5. Sample loading: inject the sample into OptiChrome through the sample valve TM In A-300. Forward centrifugation: OptiChrome TM A-300 is set to forward centrifugal rotation, set. The sample gets a spectrum through the detector, as attached ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for preparing high-purity hydroxytyrosol by using high-speed counter-current chromatography and high performance liquid chromatography in a combined manner. The method comprises the following steps of performing enzymolysis on an olive leaf extract; extracting with ethyl acetate and concentrating into hydroxytyrosol ointment of which hydroxytyrosol content is 5%-20%; mixing solvents such as petroleum ether, ethyl acetate, methanol and water proportionally; using the mixture as a high-speed counter-current solvent system under the condition that an upper phase is a fixed phase while a lower phase is a mobile phase; dissolving the hydroxytyrosol ointment in the lower phase to obtain a test article; fully filling multilayered coil separating columns of a high-speed counter-current chromatographic instrument in the fixed phase; injecting the mobile phase at revolving speed of 500-1800 r / min and at flowing speed of 1-5mL / min; and detecting by using an ultraviolet detector of which the wave length is 210-280 nanometers. Normal phase centrifugal rotation is implemented in a whole centrifugal process for 30-300 minutes. When the mobile phase begins to flow out of chromatographic columns, distillate is collected, and after the distillate is detected by HPLC (high performance liquid chromatography), the hydroxytyrosol of which the purity is 60-90% can be prepared. The method is easy to operate and low in pollution; and the purity of the separated hydroxytyrosol is high.

Description

1. Technical Field [0001] The invention relates to a method for separating hydroxytyrosol from plant leaves by enzymatic hydrolysis using high-speed countercurrent chromatography combined with high-performance liquid chromatography, and belongs to the field of natural medicine separation and medical and health care products. 2. Background technology [0002] At present, the separation technology of natural products in the world mostly adopts supercritical extraction technology, macroporous resin column chromatography technology, molecular distillation technology, membrane separation technology, etc. However, for natural products with low content and high biological activity, facing the society Advances and human requirements for high-tech refined products are extremely lacking in new technologies that can achieve separation and purification of high-purity substances and a considerable preparation capacity. If there is no such new technology, it will be difficult to realize the de...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12P7/22C07C39/11C07C37/82
Inventor 王成章原姣姣叶建中陈虹霞周昊陶冉张宇思
Owner INST OF CHEM IND OF FOREST PROD CHINESE ACAD OF FORESTRY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com