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Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges

A technology for phenolic substances and determination methods, applied in the field of analytical chemistry, can solve the problem of difficulty in detecting the real content, and achieve the effect of simple sample pretreatment and low cost

Active Publication Date: 2012-10-03
INST OF AGRI ENG TECH FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the multi-component system is measured, since the maximum absorption wavelength of each component is not the same, and the tedious pretreatment process will cause a certain loss of the substance to be measured, it is difficult to fully detect multiple groups by using the constant wavelength method. The actual content of each component in the subsystem

Method used

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  • Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges
  • Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges
  • Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Determination of grape phenols

[0023] 1 Standard sample: gallic acid, catechin, epicatechin, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, rutin, ferulic acid, benzoic acid, hesperidin, naringin, salicylic acid Acid, quercetin, resveratrol, nobiletin, orange peelin were purchased from sigma company.

[0024] 2 Sample pretreatment

[0025] 2-3g freeze-dried fruit sample (wine)→grinding with 50% ethanol→extracting twice with ethyl acetate→collecting the extract→concentrating to dryness under reduced pressure→dissolving in 2ml solvent (methanol:DMSO=1:1).

[0026] 3 Chromatographic conditions

[0027] Chromatographic column Merck LiChrospher_100RP18e (250mm×4.0mm); detection wavelength: 0-20min: 230nm; 20-35min: 280nm; 35-42min: 309nm; 42-60min: 320nm; column temperature: 30℃; gradient elution: mobile phase A is water: acetic acid = 98: 2; mobile phase B is acetonitrile. Elution program: 0min→100min, acetonitrile: 16%→70%, 2% acetic acid wat...

Embodiment 2

[0030] Embodiment 2 citrus peel pulp

[0031] 1 Standard sample: gallic acid, catechin, epicatechin, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, rutin, ferulic acid, benzoic acid, hesperidin, naringin, salicylic acid Acid, quercetin, resveratrol, nobiletin, orange peelin were purchased from sigma company.

[0032] 2 Sample pretreatment

[0033] First, separate the pulp and peel of the sample and freeze-dry it through 60 mesh for later use; mix 0.1g sample with 1mL methanol-DMSO (V / V.50:50) for 10min at room temperature, centrifuge at 9000rpm, 15min at 4°C, repeat 3 times, and combine the supernatant In a volumetric flask, dilute to volume with methanol; filter through a 0.45 μm microporous membrane for later use.

[0034] 3 Chromatographic conditions

[0035] Column Merck LiChrospher_100RP18e (250mm×4.0mm); Detection wavelength: 0-20min: 230nm; 20-35min: 280nm; 35-42min: 309nm; 42-60min: 280nm; Column temperature: 30°C; Gradient elution: mobile phase A is...

Embodiment 3

[0038] The extraction of phenolic substance in the wine sample of embodiment 3

[0039] 1 Standard sample: gallic acid, catechin, epicatechin, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, rutin, ferulic acid, benzoic acid, hesperidin, naringin, salicylic acid Acid, quercetin, resveratrol, nobiletin, orange peelin were purchased from sigma company.

[0040] 2 Sample pretreatment

[0041] Sampling and determination of monomeric phenols in wine, the extraction method is similar to that of grape fruit, the process route: wine (10 ml) → 50% ethanol extraction twice (5 times solid-liquid ratio, 70 °C extraction for 60 min) → reduced pressure concentration → Chloroform extraction 2 times→

[0042] Extract twice with ethyl acetate → concentrate under reduced pressure → extract → dissolve in methanol, dilute to 10 ml → store at 4°C for later use, repeat 3 times.

[0043] 3 Chromatographic conditions

[0044] Chromatographic column Merck LiChrospher_100RP18e (250mm...

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Abstract

The invention discloses a method utilizing the HPLC (High Performance Liquid Chromatography) to test seventeen phenol substances contained in grapes and oranges, which comprises the following steps: firstly, separating the peel and pulp of the oranges or the grapes; secondly, drying and crushing the peel and pulp, and adding DMSO (Dimethyl Sulfoxide) : methanol (1:1) or a methanol solution into the peel and pulp to execute extraction; thirdly, after the collection of supernatant fluid, repeatedly extracting and compounding the supernatant fluid to execute HPLC detection; and lastly, identifying the sorts of the grapes and the oranges according to different sorts, content analysis of various phenol substances measured in different producing areas, and different chemical compound contents. The method disclosed by the invention adopts the HPLC to synchronously separate and test the seventeen phenol substances, and achieves the test to the ingredients and contents of the seventeen chemical compounds in different sorts of fruits such as the orange and the grapes and in wine, is simple in the pretreatment of the sample and low in cost, can synchronously test the ingredients and the contents of the phenol substances in the fruits, fruit juices and fruit wine.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, in particular to a method for measuring 17 kinds of phenolic substances in grapes and citrus fruits by using HPLC. Background technique [0002] Polyphenols include simple phenols and bioflavonoids. Simple phenols include simple phenols and their derivatives and simple phenolic acids and their derivatives such as caffeic acid, chlorogenic acid, gallic acid, and the like. Bioflavonoids can be divided into two categories of compounds, one is polyphenolic monomers, that is, non-polymers, including various flavonoids and their glycosides such as rutin, quercetin, hesperidin, naringin Glycosides, nobiletin, tangeretin, etc.; the other is oligomers or polymers formed by the polymerization of monomers. Phenolic substances in fruits have a great influence on the color and flavor of fruits. Animals cannot synthesize flavonoids. Flavonoids are widely found in fruits, and are mostly distributed in th...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 陈源余亚白杨道富魏巍何志刚林晓姿谢鸿根
Owner INST OF AGRI ENG TECH FUJIAN ACAD OF AGRI SCI
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