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SPE-DES-HPLC method for simultaneously determining two flavanol substances in Shanxi mature vinegar

A technology of flavanols and aged vinegar, which is applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of complex composition of Shanxi aged mature vinegar and the inability to directly measure trace active ingredients, and achieves high sensitivity, good precision and accuracy. , the enrichment effect is good

Active Publication Date: 2021-10-29
神池县海子园醋业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that Shanxi mature vinegar contains catechin and epicatechin, but the composition of Shanxi mature vinegar is complex, and the trace active ingredients in it cannot be directly determined due to the interference of the matrix
The direct enrichment of trace active ingredients in mature vinegar and the selection of a suitable eluent to elute the active ingredients after enrichment are the key factors to be solved, but there are still few catechins and epicatechin in Shanxi mature vinegar Literature reports on content determination

Method used

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  • SPE-DES-HPLC method for simultaneously determining two flavanol substances in Shanxi mature vinegar
  • SPE-DES-HPLC method for simultaneously determining two flavanol substances in Shanxi mature vinegar
  • SPE-DES-HPLC method for simultaneously determining two flavanol substances in Shanxi mature vinegar

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: Simultaneous determination of two kinds of flavanols in Shanxi mature vinegar experimental method

[0050] 1. High performance liquid chromatography conditions

[0051] Agilent 1260 Infinity II high-performance liquid chromatography; the chromatographic column is Diamonsil C18 column, 4.6mm×250mm, 5μm; the detector is an ultraviolet detector; the flow rate is 1mL / min; the injection volume is 10μL; the column temperature is 35°C; the detection wavelength 280nm; the mobile phase is phosphate buffer and acetonitrile with a volume ratio of 0.1%; the elution method is gradient elution, as shown in Table 1 below:

[0052] Table 1 Gradient elution program

[0053]

[0054] 2. Activation of XAD-2 macroporous adsorption resin

[0055] Take 1g of macroporous resin in a beaker, add 5ml of absolute ethanol to activate for 2 hours, wash with distilled water until there is no alcohol smell, and complete the activation of the resin;

[0056] 3. Preparation of stand...

Embodiment 2

[0062] Embodiment 2: optimization of experimental conditions

[0063] 1. Single factor test optimization

[0064] 1. Selection of elution solvent

[0065] Since different eluents have different interactions with target analytes, the type of elution solvent has a great influence on the efficiency of SPE. Therefore, the present invention selects eluent: dehydrated alcohol, 70% ethanol, ethyl acetate, methyl alcohol, DES 1 (tetraethylammonium chloride: n-octanoic acid = 1:2, molar ratio), DES 2 (tetrabutylammonium chloride: n-octanoic acid = 1:2, molar ratio), DES 3 (choline chloride: acetic acid=1:2, molar ratio), according to the experimental method described in Example 1, respectively investigate its influence on the analysis effect of catechin and epicatechin.

[0066] Depend on figure 1 It can be seen that the order of the elution effects of the seven elution solvents on catechins is: ethyl acetate>DES 1 >Absolute Ethanol>Methanol>70% Ethanol>DES 3 >DES 2 ; The order...

Embodiment 3

[0100] Embodiment 3 actual vinegar sample detection

[0101] 1. Establishment of standard curve regression equation for catechin and epicatechin, limit of detection LOD (signal-to-noise ratio>3), limit of quantification LOQ (signal-to-noise ratio>10), enrichment multiple, and day-to-day relative standards Determination of deviation;

[0102] The formula for calculating the enrichment factor is as follows:

[0103]

[0104] In the formula, C sed Represents the concentration of the target in the eutectic; C 0 : initial target concentration

[0105] The mixed standard solution of each concentration prepared in Example 1 is pretreated successively according to the conditions determined in Example 2, and then the high-performance liquid chromatography conditions of Example 1 are used to detect, qualitatively by retention time, according to the measured peak of each concentration The corresponding relationship between the size of the area and its concentration draws a standar...

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Abstract

The invention belongs to the technical field of flavanol substance detection, and discloses an SPE-DES-HPLC method for simultaneously determining two flavanol substances in Shanxi mature vinegar. According to the method, a Dimonsil C18 chromatographic column is selected, and the flow velocity is 1mL / min; the sample size is 10 [mu] L; the detection wavelength is 280 nm; a mobile phase is a 0.1% phosphate buffer and acetonitrile; and gradient elution is carried out. Through an optimization experiment, when the dosage of the activated XAD-2 macroporous adsorption resin is finally determined to be 188mg, the volume of the eutectic solvent (tetraethylammonium chloride-n-caprylic acid in a molar ratio of 1: 3) is 400mu L, the adsorption time is 11min, and the analysis time is 20min, the enrichment effect of the two flavanol substances is optimal. Under the condition, the linear relation of catechin and epicatechin is sound (R2 > 0.99), and compared with the tedious process of traditional solid-phase extraction, the detection limit, the quantitation limit, the intraday and interday relative standard deviation and the adding standard recovery rate show that the method is simpler, more convenient, high in sensitivity and small in matrix interference, and is suitable for enrichment determination of catechin and epicatechin in Shanxi mature vinegar.

Description

technical field [0001] The invention belongs to the technical field of detection of flavanols, in particular to a SPE-DES-HPLC method for simultaneously measuring two kinds of flavanols in Shanxi aged vinegar. Background technique [0002] Catechin and epicatechin are a class of active flavanols mainly found in natural plants such as tea. Studies have shown that catechins have anti-oxidation, anti-inflammatory, anti-obesity, anti-bacterial, anti-viral, reduce bone loss, prevent food poisoning, promote the absorption of intestinal active substances, reduce the rate of influenza infection, prevent dental caries and other oral diseases, prevent The efficacy of cardiovascular disease and cancer, protection of red blood cells from permanganate-induced hemolysis, red blood cell protein sulfhydryl oxidation and membrane lipid peroxidation, and even the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease. Catechin will be metabolized rapidly when it e...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/86
CPCG01N30/02G01N30/06G01N30/08G01N30/8679
Inventor 白宝清范三红张锦华杨钰昆
Owner 神池县海子园醋业有限公司
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