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Low temperature sample introduction method capable of controlling proportion of punicalagin isomers

A technology for punicalagin and isomers, which is applied in the field of low temperature sampling that can control the ratio of punicalagin isomers

Inactive Publication Date: 2019-10-11
XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no literature on the artificial control of the ratio of α-punicalagin and β-punicalagin in punicalagin to a fixed ratio

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Weigh 1 mg of pomegranate peel extract with a punicalagin content of about 32% and a ratio of α-punicalagin to β-punicalagin of 42:58, dissolve it in 1 mL of methanol, and heat it in a water bath at 70°C for 1 hour to Dry, add 1mL water to dissolve, pass through a 0.45μm organic filter head, place at 0°C for 1h, and mark it as sample 1;

[0012] Take an analytical reversed-phase C18 chromatographic column, the specification column length is 250mm×inner diameter 4.6mm, the filler particle size is 10μm, the mobile phase is methanol-0.1% formic acid water with a volume ratio of 12:88, the flow rate is 0.6min / mL, and the temperature is 30°C. The ultraviolet detection wavelength is 254nm, equilibrate for 15min, and wait for use; take 20μL of sample 1 and inject it into a well-balanced chromatographic column, among which, the peak between 12.5min and 15min is α-punicalagin, and the peak between 27min and 32.5min is β-Punicalagin. Determination of α-punicalagin (A α ) and β-...

Embodiment 2

[0014] Weigh 1 mg of pomegranate peel extract with a punicalagin content of about 32%, and a ratio of α-punicalagin to β-punicalagin of 42:58, dissolve it in 1 mL of water, pass through a 0.45 μm organic filter head, and heat at 4 Place it overnight in the refrigerator at 0°C for 30 minutes at a temperature of 0°C, and mark it as sample 2;

[0015] Take an analytical reversed-phase C18 chromatographic column, the specification column length is 250mm×inner diameter 4.6mm, the filler particle size is 10μm, the mobile phase is methanol-0.1% formic acid water with a volume ratio of 12:88, the flow rate is 0.6min / mL, and the temperature is 30°C. The ultraviolet detection wavelength is 254nm, equilibrate for 15min, and wait for use; take 20μL of sample 2 and inject it into a well-balanced chromatographic column, among which, the peak between 12.5min and 15min is α-punicalagin, and the peak between 27min and 32.5min is β-punicalagin, determination of which α-punicalagin (A α ) and β...

Embodiment 3

[0017] Weigh 1 mg of pomegranate peel extract with a punicalagin content of about 32%, and a ratio of α-punicalagin to β-punicalagin of 42:58, dissolve it in 1 mL of water, pass through a 0.45 μm organic filter head, and heat at 4 ℃ overnight in the refrigerator, at a temperature of 10 ℃ for 5 hours, marked as sample 3;

[0018] Take an analytical reversed-phase C18 chromatographic column, the specification column length is 250mm×inner diameter 4.6mm, the filler particle size is 10μm, the mobile phase is methanol-0.1% formic acid water with a volume ratio of 12:88, the flow rate is 0.6min / mL, and the temperature is 30°C. The ultraviolet detection wavelength is 254nm, equilibrate for 15min, and wait for use; take 20μL of sample 3 and inject it into the well-balanced chromatographic column, among which, the peak between 12.5min and 15min is α-punicalagin, and the peak between 27min and 32.5min is β-punicalagin, determination of which α-punicalagin (A α ) and β-punicalagin (A β...

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Abstract

The invention relates to a low temperature sample introduction method capable of controlling the proportion of punicalagin isomers. According to the method, on the basis of the isomerization kineticsand thermodynamics study between two epimers of punicalagin, after a pomegranate bark extract containing rich punicalagin is respectively isomerized to stable proportions in methanol, ethanol, isopropanol, acetonitrile, methanol-water or water, a sample is introduced into a liquid chromatogram to perform separation and analysis, the isomerization kinetics process is prevented by applying a low temperature environment in a sample introduction process, so that the punicalagin maintains the isomer proportion prior to the sample introduction on a chromatographic column. The result shows that thelow temperature sample introduction technique can effectively fix the proportion of alpha-punicalagin and beta-punicalagin in the isomerized pomegranate bark extract to 32-50:50-68. By adopting the method, the alpha-punicalagin and the beta-punicalagin can be manually increased and is not decreased by isomerization in achromatographic separation process, so that the yield of the single isomer alpha-punicalagin and the beta-punicalagin in a preparation process is improved.

Description

technical field [0001] The invention relates to a low-temperature sample injection method capable of controlling the ratio of punicalagin isomers. The method is based on the isomerization kinetics and thermodynamics research between the two epimers of punicalagin. After the pomegranate peel extract is isomerized into a stable ratio in the solvent, it is then injected into the liquid chromatography for separation and analysis. By applying a low temperature environment during the injection process to prevent its isomerization kinetics, so that punicalagin Pre-injection isomer ratios are maintained on the column. The method can maintain the isomerization ratio of punicalagin isomers in the process of chromatographic separation, can increase the artificial control of α-punicalagin or β punicalagin without being affected by isomerization in the process of chromatographic separation Reduction can improve the yield in the preparation process of single isomer α-punicalagin and β-puni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/16G01N30/06
CPCG01N30/06G01N30/16
Inventor 阿吉艾克拜尔·艾萨孙光映赵永昕木尼热·阿布都艾尼古丽契热·阿地力
Owner XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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