Method for preparing chemical reference substances of niga-ichigoside F1 and suavissimoside R1 simultaneously

Abstract

The invention relates to a method for preparing chemical reference substances of niga-ichigoside F1 and suavissimoside R1 simultaneously. The method comprises the following steps of extracting crude extract from a rubus parvifolius L. medicinal material or adopting directly rubus parvifolius L. extract as a raw material, and carrying out high-efficiency purification through two processes of resin column separation and high speed counter current chromatography to obtain the chemical reference substances of niga-ichigoside F1 and suavissimoside R1, wherein purities of the chemical reference substances are above 98%. The method has the advantages of simple processes, high product purity and good adaptability for mass production.

Description

technical field

[0001] The present invention belongs to the technical field of medicine, and in particular relates to a method for simultaneously preparing niga-ichigoside F1 and suavissimoside R1 chemical reference substances by using resin column separation and high-speed countercurrent chromatography. Background technique

[0002] Mao berry ( Rubus parvifolius L.) is the dried whole herb of the Rubus genus Rubus in the family Rosaceae. It is a commonly used herbal medicine in the folk. It has the functions of clearing away heat and detoxification, dispelling blood stasis and stopping bleeding, expelling wind and dampness, promoting blood circulation and cooling blood, killing insects and treating sores, etc. It is used in gynecology to stop bleeding ,Significant effect. Modern studies have shown that the extract of Mao berry has a protective effect on cerebral ischemia, has the effect of hemostasis and promoting blood circulation and removing blood stasis, and has the s...

Examples

Embodiment 1

[0019] 1. Coarse separation of resin column: take 10g of the extract of Mao berry as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on a HPD-100 macroporous resin column, first wash with 5BV of water to remove impurities, and then use 4 -7BV was eluted with 50% ethanol solution, the eluate was collected, concentrated and dried to obtain 408 mg of crude product;

[0020] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 4:5:2) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the main engine to rotate, the speed is controlled at 1000r / min, and at the same time Pump the mobile phase at...

Embodiment 2

[0023] 1. Coarse separation with resin column: take 50g of the chrysanthemum berry extract as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on the AB-8 macroporous resin column, first wash with 4BV of water to remove impurities, and then use 5BV50 % ethanol solution was eluted, and the eluate was collected, concentrated and dried to obtain 2.2g of crude product;

[0024] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 7: 8:2) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the host to rotate, the speed is controlled at 750r / min, and at the same time Pump the mobile phase at a fl...

Embodiment 3

[0027] 1. Coarse separation by resin column: take 50g of the extract of Mao berry as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on HPD-100 macroporous resin column, first wash with 5BV of water to remove impurities, and then use 4BV50 % ethanol solution was eluted, and the eluate was collected, concentrated and dried to obtain 2.3g of crude product;

[0028] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 7:5:5) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the main engine to rotate, the speed is controlled at 700r / min, and at the same time Pump the mobile phase at a flow r...

technical field

[0001] The present invention belongs to the technical field of medicine, and in particular relates to a method for simultaneously preparing niga-ichigoside F1 and suavissimoside R1 chemical reference substances by using resin column separation and high-speed countercurrent chromatography. Background technique

[0002] Mao berry ( Rubus parvifolius L.) is the dried whole herb of the Rubus genus Rubus in the family Rosaceae. It is a commonly used herbal medicine in the folk. It has the functions of clearing away heat and detoxification, dispelling blood stasis and stopping bleeding, expelling wind and dampness, promoting blood circulation and cooling blood, killing insects and treating sores, etc. It is used in gynecology to stop bleeding ,Significant effect. Modern studies have shown that the extract of Mao berry has a protective effect on cerebral ischemia, has the effect of hemostasis and promoting blood circulation and removing blood stasis, and has the s...

Examples

Embodiment 1

[0019] 1. Coarse separation of resin column: take 10g of the extract of Mao berry as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on a HPD-100 macroporous resin column, first wash with 5BV of water to remove impurities, and then use 4 -7BV was eluted with 50% ethanol solution, the eluate was collected, concentrated and dried to obtain 408 mg of crude product;

[0020] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 4:5:2) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the main engine to rotate, the speed is controlled at 1000r / min, and at the same time Pump the mobile phase at...

Embodiment 2

[0023] 1. Coarse separation with resin column: take 50g of the chrysanthemum berry extract as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on the AB-8 macroporous resin column, first wash with 4BV of water to remove impurities, and then use 5BV50 % ethanol solution was eluted, and the eluate was collected, concentrated and dried to obtain 2.2g of crude product;

[0024] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 7: 8:2) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the host to rotate, the speed is controlled at 750r / min, and at the same time Pump the mobile phase at a fl...

Embodiment 3

[0027] 1. Coarse separation by resin column: take 50g of the extract of Mao berry as raw material, dissolve it with 15% ethanol solution, filter, and separate the filtrate on HPD-100 macroporous resin column, first wash with 5BV of water to remove impurities, and then use 4BV50 % ethanol solution was eluted, and the eluate was collected, concentrated and dried to obtain 2.3g of crude product;

[0028] 2. Purification by high-speed countercurrent chromatography: use dichloromethane-methanol-water system as a two-phase solvent system, put the system dichloromethane-methanol-water (volume ratio 7:5:5) in a separatory funnel at room temperature Shake, place, and balance for a period of time, take the upper phase as the stationary phase, and the lower phase as the mobile phase, pump the stationary phase into the chromatographic column to fill the column, then start the main engine to rotate, the speed is controlled at 700r / min, and at the same time Pump the mobile phase at a flow r...