# US 9,517,998 B1 · Device method of making, separating, and purifying artepillin C in propolis

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 9,517,998 B1
- Document ID: US9517998B1
- Status: Granted
- Kind: B1
- Application number: 14/965,965
- Filed: 2015-12-11
- Granted: 2016-12-13
- Earliest claimed: 2015-09-08
- Assignees: Ming-Hsi Chuang
- Applicants: Ming-Hsi Chuang
- Inventors: Ming-Hsi Chuang; Chi-An Li; Zer-Ran Yu; Shuen-Ane Kang; Chu-Ting Liu; Lin-Hsiang Chuang; Chiu-Ying Peng; I-Lung Yu; Cheng-Yu Chi; Hsin-Yu Chung
- Web page: https://patentgrep.com/patent/US9517998B1

## CPC

- C07C 51/47: by solid-liquid treatment; by chemisorption
- B01D 15/265: {Adsorption chromatography}
- B01D 15/40: using supercritical fluid as mobile phase or eluent
- Y02P 20/54: using solvents, e.g. supercritical solvents or ionic liquids
- C07C: ACYCLIC OR CARBOCYCLIC COMPOUNDS
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

A method of making, separating, and purifying artepillin C in propolis includes mixing propolis with ethanol to obtain a propolis-ethanol extract; providing supercritical carbon dioxide and the propolis-ethanol extract to a chromatographic column to separate wax, artepillin C, and flavonoids; providing supercritical carbon dioxide, the artepillin C, and the flavonoids to an adsorption column to remove the flavonoids; keeping the adsorption column still; and providing ethanol to the adsorption column to obtain the purified artepillin C.

## Claims

### Claim 1

A method of obtaining artepillin C in propolis, comprising the steps of:
mixing propolis with ethanol to form a mixture, and then removing impurities of the mixture to obtain a propolis-ethanol extract;
providing supercritical carbon dioxide and the propolis-ethanol extract to a chromatographic column to separate wax, artepillin C, and flavonoids from the propolis-ethanol extract, and then removing the wax at a bottom of the chromatographic column, and collecting the artepillin C and the flavonoids at a top of the chromatographic column;
providing supercritical carbon dioxide, the artepillin C, and the flavonoids to an adsorption column, and then removing the flavonoids at a bottom of the adsorption column, wherein the artepillin C is adsorbed by the adsorption column;
keeping the adsorption column still for a predetermined time; and
providing ethanol to the adsorption column to separate the artepillin C from the adsorption column, and then collecting the artepillin C at a top of the adsorption column.

### Claim 2 (depends on 1)

The method of claim 1, wherein a ratio of the propolis and the ethanol in the mixture is 1:5-10 (W/V), and a volume of the mixture is 10-20 L.

### Claim 3 (depends on 2)

The method of claim 2, wherein a weight of the propolis for the mixture is in a range between 1 Kg and 2 Kg.

### Claim 4 (depends on 1)

The method of claim 1, wherein the chromatographic column is set to 3,000-4,000 psi and 40-60° C., a flow rate of the supercritical carbon dioxide is set to 6-9 L/hr, and a flow rate of the propolis-ethanol extract is set to 1-3 L/hr for providing the supercritical carbon dioxide and the propolis-ethanol extract to the chromatographic column.

### Claim 5 (depends on 4)

The method of claim 4, wherein the chromatographic column has a stainless container with 36 mm in an interior diameter and 1,000 mm in a height; a stainless plate is received in the container, and the plate is made of a material selected from the group consisting of Pro-Pak protruded metal, saddles, rings, structured packing, and knitted packing.

### Claim 6 (depends on 1)

The method of claim 1, wherein the adsorption column is set to 2,000-3,000 psi and 40-60° C., and a flow rate of the supercritical carbon dioxide is set to 6-9 L/hr for providing the supercritical carbon dioxide to the adsorption column.

### Claim 7 (depends on 6)

The method of claim 6, wherein the adsorption column has a stainless container with 36 mm in an interior diameter and 1,000 mm in a height, and the stainless container receives a sorbent, wherein the sorbent is selected from the group consisting of silica gel, sephadex, and resin.

### Claim 8 (depends on 1)

The method of claim 1, wherein the adsorption column is kept still for 1-2 hr.

### Claim 9 (depends on 1)

The method of claim 1, wherein the adsorption column is set to 2,000-3,000 psi and 40-60° C., and a flow rate of the ethanol is 1-3 L/hr for providing the ethanol to the adsorption column.