Method for extracting chlorogenic acid and neochlorogenic acid from bunge pricklyash leaves

Abstract

The invention discloses a method for extracting chlorogenic acid and neochlorogenic acid from bunge pricklyash leaves. The method is characterized by comprising the following steps of: crushing dried bunge pricklyash leaves till the average particle size is 400-600 micrometers; adding to methanol with the concentration of 50wt%-80wt% according to the material-to-liquid ratio of 1g:(8-30)mL, oscillating at room temperature for 24-36 hours, separating supernatant, concentrating, and drying to obtain a methanol extract; dispersing the methanol extract into distilled water at the material-to-ratio of 1g:(10-30)mL, adding ethyl acetate according to the volume ratio of 1:2, carrying out oscillating extraction for 2-3 times, collecting a water-phase extractive, concentrating and drying, wherein the yield is 19.8%-25.8%; carrying out gradient elution on a reveres-phase resin SciBioChem MCI-GEL chromatographic column positioned on a dried sample by using methanol-water, wherein the filler height is 20cm, the column diameter is 3.2 cm, the concentrations of methanol-water are 0wt%, 10wt%, 20wt%, 30wt%, 40wt% and 50wt%, and the elution flow velocity is1-2mL / min; respectively collecting 10wt% methanol eluent and 20wt% methanol eluent, concentrating, drying, and then separating and purifying by using liquid chromatography to obtain the neochlorogenic acid and the chlorogenic acid, wherein the yield is 0.03%-0.05% and 0.03%-0.06% respectively.

Description

technical field

[0001] The invention relates to a method for preparing chlorogenic acid and neochlorogenic acid from prickly ash leaves, and belongs to the technical field of Chinese herbal medicine preparation and chemical industry. Background technique

[0002] Chlorogenic acid is a depsipolic acid composed of caffeic acid and quinic acid, also known as coffee tannic acid, which is a kind of plant produced by shikimic acid pathway during aerobic respiration. A phenylpropanoid compound. It widely exists in plants, and its content is higher in honeysuckle and Eucommia. Chlorogenic acid is an important biologically active substance with a wide range of pharmacological effects and biological activities. It is not only the main active ingredient of many medicinal materials and Chinese patent medicines for antibacterial detoxification, anti-inflammatory and choleretic, but also has anti-virus, white blood cell increase, anti-tumor, Effects of lowering blood pressure, lowering ...

Examples

Embodiment 1

[0039] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 600 μm, then use 300 mL of methanol with a concentration of 80 wt % as an extractant, shake at room temperature for 36 hours, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0040](2) Disperse the above-mentioned methanol extract in 90ml distilled water, the ratio of solid to liquid is 1g: 30mL, then use 180mL ethyl acetate to shake and extract twice, collect the aqueous phase extract, concentrate and dry, and obtain 2.58g of dried sample, the yield 25.8%;

[0041] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow rate i...

Embodiment 2

[0046] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 500 μm, then use 200 mL of methanol with a concentration of 70 wt % as an extractant, shake at room temperature for 30 h, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0047] (2) Disperse the above-mentioned methanol extract in 50ml of distilled water, the ratio of solid to liquid is 1g: 20mL, then use 100mL of ethyl acetate to shake and extract twice, collect the aqueous phase extract, concentrate and dry, and obtain 2.12g of the dried sample, the yield 21.2%;

[0048] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow...

Embodiment 3

[0053] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 500 μm, then use 100 mL of methanol with a concentration of 60 wt % as an extractant, shake at room temperature for 24 hours, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0054] (2) Disperse the above-mentioned methanol extract in 25ml of distilled water, the ratio of solid to liquid is 1g: 10mL, then shake and extract once with 25mL of ethyl acetate, collect the aqueous phase extract, concentrate and dry, and obtain 2.07g of dried sample, the yield 20.7%;

[0055] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow rat...

technical field

[0001] The invention relates to a method for preparing chlorogenic acid and neochlorogenic acid from prickly ash leaves, and belongs to the technical field of Chinese herbal medicine preparation and chemical industry. Background technique

[0002] Chlorogenic acid is a depsipolic acid composed of caffeic acid and quinic acid, also known as coffee tannic acid, which is a kind of plant produced by shikimic acid pathway during aerobic respiration. A phenylpropanoid compound. It widely exists in plants, and its content is higher in honeysuckle and Eucommia. Chlorogenic acid is an important biologically active substance with a wide range of pharmacological effects and biological activities. It is not only the main active ingredient of many medicinal materials and Chinese patent medicines for antibacterial detoxification, anti-inflammatory and choleretic, but also has anti-virus, white blood cell increase, anti-tumor, Effects of lowering blood pressure, lowering ...

Examples

Embodiment 1

[0039] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 600 μm, then use 300 mL of methanol with a concentration of 80 wt % as an extractant, shake at room temperature for 36 hours, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0040](2) Disperse the above-mentioned methanol extract in 90ml distilled water, the ratio of solid to liquid is 1g: 30mL, then use 180mL ethyl acetate to shake and extract twice, collect the aqueous phase extract, concentrate and dry, and obtain 2.58g of dried sample, the yield 25.8%;

[0041] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow rate i...

Embodiment 2

[0046] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 500 μm, then use 200 mL of methanol with a concentration of 70 wt % as an extractant, shake at room temperature for 30 h, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0047] (2) Disperse the above-mentioned methanol extract in 50ml of distilled water, the ratio of solid to liquid is 1g: 20mL, then use 100mL of ethyl acetate to shake and extract twice, collect the aqueous phase extract, concentrate and dry, and obtain 2.12g of the dried sample, the yield 21.2%;

[0048] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow...

Embodiment 3

[0053] (1) Grind 10 g of dried Zanthoxylum bungeanum leaves, with an average particle size of 500 μm, then use 100 mL of methanol with a concentration of 60 wt % as an extractant, shake at room temperature for 24 hours, separate the supernatant, and concentrate and dry the supernatant to obtain methanol extract ;

[0054] (2) Disperse the above-mentioned methanol extract in 25ml of distilled water, the ratio of solid to liquid is 1g: 10mL, then shake and extract once with 25mL of ethyl acetate, collect the aqueous phase extract, concentrate and dry, and obtain 2.07g of dried sample, the yield 20.7%;

[0055] (3) Take 1 g of the above-mentioned dried sample, dissolve it in 10 mL of distilled water, put it on a reversed-phase resin SciBioChem MCI-GEL chromatography column, the filler height is 20 cm, and the column diameter is 3.2 cm, and it is eluted with methanol-water gradient, and the concentration is 0 wt%. , 10wt%, 20wt%, 30wt%, 40wt%, 50wt% methanol, the elution flow rat...

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