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Industrial high performance liquid chromatography refining method of stevioside St

A technology of stevioside and chromatographic system, which is applied in the field of preparation of stevioside, can solve the problems of long time and low purity, and achieve the effect of easy control and simplified process technology

Inactive Publication Date: 2014-05-14
JIANGSU HANBON SCI & TECH CO
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide a new method for preparing high-purity steviol glycoside St by using an industrial preparative liquid chromatography separation system to solve the problems of long time and low purity in the process of refining stevioside crude products. Include the following steps:

Method used

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  • Industrial high performance liquid chromatography refining method of stevioside St
  • Industrial high performance liquid chromatography refining method of stevioside St
  • Industrial high performance liquid chromatography refining method of stevioside St

Examples

Experimental program
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Effect test

Embodiment 1

[0013] 1. Take 400mg of steviol glycoside crude product and mix it with aqueous solution, configure it into a saturated solution, place it in a filter device, and filter to remove solid particles;

[0014] 2. Inject the steviol glycoside solution into the dynamic axial compression column to prepare the chromatographic system. The dynamic axial compression column size is Φ50×250mm, the octadecyl reversed-phase silica gel particle size is 10μm, the sample volume is 400mg, and the flow rate of the mobile phase is 80ml / min. In the initial stage of separation, the volume ratio of mobile phase methanol to water ranges from 30 / 70 to 20 / 80. After 45 minutes of elution, the volume ratio of mobile phase methanol to water is switched to 90 / 10, and the volume ratio of mobile phase methanol to water is eluted to 65 minutes to wash out the rear-end impurities. , a separation cycle ends. The detection wavelength of the ultraviolet photometric detector that adopts is 210nm, collects the cut...

Embodiment 2

[0016] 1. Take 1g of steviol glycoside crude product and mix it with aqueous solution, configure it into a saturated solution, place it in a filter device, and filter to remove solid particles;

[0017] 2. Inject the steviol glycoside solution into the dynamic axial compression column to prepare the chromatographic system. The dynamic axial compression column size is Φ50×250mm, the octadecyl reversed-phase silica gel particle size is 10μm, the sample volume is 1g, and the flow rate of the mobile phase is 80ml / min. In the initial stage of separation, the volume ratio of mobile phase methanol to water is 30 / 70~20 / 80. After 45 minutes of elution, the volume ratio of mobile phase methanol to water is switched to 90 / 10, and the volume ratio of mobile phase methanol to water is 90 / 10. The cycle ends. The detection wavelength of the ultraviolet photometric detector that adopts is 210nm, collects the cut that retention time is in 29~33min, as image 3 Shown, through HPLC analysis p...

Embodiment 3

[0019] 1. Take 1g of steviol glycoside crude product and mix it with aqueous solution, configure it into a saturated solution, place it in a filter device, and filter to remove solid particles;

[0020] 2. Inject the steviol glycoside solution into the dynamic axial compression column to prepare the chromatographic system. The dynamic axial compression column size is Φ50×250mm, the octadecyl reversed-phase silica gel particle size is 20~45μm, the sample volume is 1g, and the flow rate of the mobile phase is It is 80ml / min. In the initial stage of separation, the volume ratio of mobile phase methanol to water is 40 / 60~20 / 80. After 55 minutes of elution, the volume ratio of mobile phase methanol to water is switched to 90 / 10, and the volume ratio of mobile phase methanol to water is 90 / 10. After elution for 65 minutes, the back-end impurities are washed out. The cycle ends. The detection wavelength of the ultraviolet photometric detector that adopts is 210nm, collects the fract...

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Abstract

The invention belongs to the natural products extraction field, and employs a stevia rebaudiana extract product crude product as a raw material, an appropriate moving phase and column separating condition is obtained through a HPLC optimization condition, and is amplified to industrial preparation scale liquid chromatogram separating system, and stevioside St with high purity is refined. The invention has the advantages that the crude product is used for preparing the required substance by using industrial preparation scale liquid chromatogram separation, the good separating and purification effect can be reached, the solvent has the advantages of low cost, simple operation, short period and high efficiency, and the purity can reach more than 99.5%.

Description

technical field [0001] The invention belongs to the field of natural product extraction, and in particular relates to a preparation method of steviol glycosides. Background technique [0002] Stevia, also known as stevia sugar, is an extract of stevia rebaudiana leaves, which does not contain sugar and calories. They are a class of sweet terpene glycosides, white to slightly yellow in color, suitable in taste, and have no peculiar smell. They are new sugar sources with broad development prospects. Among them, steviol glycoside St accounts for 60%~70% of the total glycosides, and its sweetness is 300 times that of sucrose. It is a sweetener approved by the Ministry of Health of my country. It has a natural low calorific value and is very close to the taste of sucrose. It is the third natural sweetener with development value and health praise after sugarcane and beet sugar, and is known internationally as "the third sugar source in the world". Steviol glycoside products can ...

Claims

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Application Information

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IPC IPC(8): C07H15/256C07H1/06
Inventor 张宇王亚辉沈健增
Owner JIANGSU HANBON SCI & TECH CO