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A kind of industrial grade preparation liquid chromatography separation method of mogroside V

A mogroside and liquid phase preparation technology, applied in chemical instruments and methods, glycoside steroids, steroids, etc., can solve the problems affecting the use efficiency and life of the prepared liquid phase filler, unsuitable for industrial production, complicated processing process, etc. , to achieve the effect of stable and reliable product quality, high degree of automation and simple process flow

Active Publication Date: 2017-05-31
HUNAN HUACHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is not suitable for industrial production due to the complicated process, low yield and high cost due to repeated extraction and chromatography.
[0007] CN104086614A discloses a preparation method of Luo Han Guo extract suitable for industrial production. The method is to crush fresh Luo Han Guo, extract, concentrate, centrifuge, ultrafiltration, multi-stage adsorption and separation of macroporous resin, and perform nanofiltration, concentration and drying. Finally, four products with different specifications can be produced at the same time, and the total yield of mogroside V is >70%. This method is suitable for industrial production, but the purity of the obtained product is low, and the subsequent treatment process is relatively complicated
[0009] CN104530168A discloses an industrialized preparation method of Mogroside V. The method is to extract and filter the Luo Han Guo, use a preparative liquid chromatography system to separate, concentrate and dry the product, and the product purity can reach more than 95%. The process of the method is simple. The product has high purity, but because it has not been systematically filtered and purified before being put on the column, it seriously affects the service efficiency and life of the prepared liquid phase packing, the production cost is high, and it is still in the test scale-up stage, and there is no example of production scale-up

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) Dissolution and filtration: Take 150g of the crude product of Luo Han Guo (the content of mogroside V is 25%), dissolve it with 2500 mL of methanol, and filter it with a 200-mesh stainless steel mesh to obtain the filtrate;

[0036] (2) Centrifugation and ultrafiltration: The filtrate obtained in step (1) is subjected to tubular centrifugation at a centrifugal rate of 14,000r / min, and then a second ultrafiltration is performed using a roll-type ultrafiltration membrane. The molecular weight cut-off of the ultrafiltration membrane is 30,000 respectively. Daltons and 4000 Daltons, get 2250mL ultrafiltrate;

[0037] (3) Preparative liquid chromatography separation: inject the ultrafiltrate obtained in step (2) into the dynamic axial compression preparative liquid chromatography system through the injection pump, the injection volume is 2250 mL, advanced pre-column treatment, the specification of the pre-column is Ф200×50mm, the packing is octadecyl reversed-phase silic...

Embodiment 2

[0045] (1) Dissolution and filtration: take 150g of the crude product of Luo Han Guo (mogroside V content is 25%), dissolve with 2300mL of methanol, and filter with 200 mesh stainless steel mesh to obtain the filtrate;

[0046] (2) Centrifugation and ultrafiltration: The filtrate obtained in step (1) is subjected to tubular centrifugation at a centrifugal rate of 15,000r / min, and then a second ultrafiltration is performed using a roll-type ultrafiltration membrane, and the molecular weight cut-off of the ultrafiltration membrane is 50,000. Dalton and 6000 Dalton, get ultrafiltrate 2125mL;

[0047] (3) Preparative liquid chromatography separation: inject the ultrafiltrate obtained in step (2) into the dynamic axial compression preparative liquid chromatography system through the injection pump, the injection volume is 2125 mL, advanced pre-column treatment, the specification of the pre-column is Ф200 ×50mm, the filler is octadecyl reversed-phase silica gel, the particle size of t...

Embodiment 3

[0055] (1) Dissolving and filtering: Take 150g of the crude product of Luo Han Guo (the content of mogroside V is 25%), dissolve it with 1850 mL of methanol, and filter it with a 200-mesh stainless steel mesh to obtain the filtrate;

[0056] (2) Centrifugation and ultrafiltration: The filtrate obtained in step (1) is subjected to tubular centrifugation at a centrifugal rate of 12,000r / min, and then a second ultrafiltration is performed using a roll-type ultrafiltration membrane. The molecular weight cut-off of the ultrafiltration membrane is 40,000 respectively. Dalton and 5000 Dalton, get ultrafiltrate 1715mL;

[0057] (3) Preparative liquid chromatography separation: inject the ultrafiltrate obtained in step (2) into the dynamic axial compression preparative liquid chromatography system through the injection pump, the injection volume is 1715mL, advanced pre-column treatment, the specification of the pre-column is Ф200 ×50mm, the filler is octadecyl reversed-phase silica gel...

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Abstract

An industrial-grade preparative liquid chromatographic separation method of mogroside V comprises steps as follows: (1) dissolution and filtration: adding methanol to a crude Siratia grosvenorii product in the solid-to-liquid ratio of 1:(10-20) for dissolution, and performing filtration to obtain a filtrate; (2) centrifugation and ultrafiltration: centrifuging the filtrate obtained in Step (1), and then performing ultrafiltration to obtain an ultrafiltrate; (3) preparative liquid chromatographic separation: injecting the ultrafiltrate obtained in Sep (2) into a dynamic axial compression preparative liquid chromatographic system, then performing elution with an eluent, and collecting a target-section eluent; (4) concentration and drying: concentrating the target-section eluent obtained in Step (3), and then performing drying to obtain the mogroside V. The method is simple in technological process, high in automation degree, high in controllability and high in refining degree, the product quality reproducibility is good, and the product quality is stable and reliable; the purity of the mogroside V separated with the method is higher than or equal to 95%, and the yield is higher than or equal to 75%.

Description

technical field [0001] The invention relates to a separation method of mogroside V, in particular to an industrial-grade liquid chromatography separation method of mogroside V. Background technique [0002] Luo Han Guo, Latin scientific name Siratia grosvenorii, alias La Han Guo, fake bitter gourd, is native to Guilin, Guangxi, and Yongfu County and Longsheng County are rich in it, and its quality is high. Currently, it is also planted in Hunan, Jiangxi, Guangdong and other provinces. [0003] Luo Han Guo is a plant with the same origin of medicine and food. It can be used to make drinks and medicine. It is rich in nutrition and extremely sweet. Studies have shown that the main sweet substances in Luo Han Guo are mogrosides, and the most important chemical component is mogroside V, which has a sweetness of more than 250 times that of sucrose, and its calories are only 20 times that of sucrose. one. [0004] Mogroside V is a natural sweetener, which has the characteristics ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07J17/00
CPCC07J17/005
Inventor 唐方华黄华学
Owner HUNAN HUACHENG BIOTECH