Method of on-line quickly detecting total anti-oxidizing property of sample

A technology for anti-oxidation and detection of samples, applied in the field of detection and analysis, can solve the problem that the antioxidant activity of the total mixture cannot be directly and quickly measured

Active Publication Date: 2016-01-13
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the HPLC-DPPH / ABTS method currently reported is characterized by online analysis of the antioxidant activity of a single compound, and cannot directly and quickly determine the antioxidant activity of the total mixture.
There is no report on the total antioxidant activity of HPLC-on-line rapid detection samples

Method used

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  • Method of on-line quickly detecting total anti-oxidizing property of sample
  • Method of on-line quickly detecting total anti-oxidizing property of sample
  • Method of on-line quickly detecting total anti-oxidizing property of sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Preparation of standard stock solution

[0043] Accurately weigh 25.00mg of rutin, gallic acid, ferulic acid, caffeic acid and chlorogenic acid in a 25mL brown volumetric flask, dissolve with 80% methanol and dilute to 25mL as a standard stock solution (1.00mgmL -1 ). Prepare a series of standard solutions by serial dilution.

[0044] Vitamin C water standard solution is prepared with chromatographic grade double distilled water, and needs to be prepared and used immediately.

[0045] 2. Configuration of free radical solution

[0046] Preparation of DPPH free radical solution: Weigh DPPH powder into 500mL methanol solution, and prepare DPPH free radical methanol solutions with concentrations of 0.1mM, 0.2mM and 0.5mM respectively.

[0047] ABTS radical reaction (ABTS · + ) solution configuration: 5mL ABTS solution + 88 μL potassium superphosphate aqueous solution, left overnight. Take 1mLABTS · + About 100mL of absolute ethanol was added to the stock solution, ...

Embodiment 2

[0058] 1. Sample processing

[0059] After the citrus fruit is cleaned, the citrus fruit is divided into four parts: citrus peel, sac, seed and juice. The juice is obtained by manually squeezing the citrus sac valves wrapped in three layers of gauze. The citrus solid part is placed in 50 DEG C of blast drying ovens and dries for 2-3 days. Crush the dried samples of citrus peels, pass through a 60-mesh sieve (0.280 mm), store them in a desiccator at room temperature, and wait for analysis. Accurately weigh 250 mg of citrus dry powder sample, 625 mg of seeds or 2 mL of citrus juice in a 50 mL centrifuge tube, add 80% methanol to a volume of 25 mL, mix thoroughly, ultrasonically extract at 55°C for 30 min, and shake for 12 h. Afterwards, the sample was centrifuged at a speed of 5000g for 10min, and the supernatant was taken and stored at -80°C. When analyzing citrus extracts on-line by HPLC-FRSD, it is necessary to filter the supernatant through a 0.22 micron organic filter me...

Embodiment 3

[0070] 1. Linear relationship investigation and determination of detection limit and quantification limit

[0071] Precisely draw the stock solution of rutin, gallic acid, ferulic acid, caffeic acid and chlorogenic acid into five 25mL brown volumetric flasks respectively. Using the gradual dilution method, a series of standard solutions with different concentrations were prepared. According to the "HPLC-FRSD online antioxidant method condition optimization", the finally optimized and established analysis conditions were analyzed online, and 25 μL of each standard solution was injected separately. Take the concentration of phenolic standard solution as the abscissa (X) and the negative peak area as the ordinate (Y), draw a standard curve and calculate the regression equation and correlation coefficient.

[0072] The standard solution was diluted by gradual dilution method, and when the signal-to-noise ratio (RSN) was set to 3, the lower limit of detection (LOD) of each standard...

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Abstract

The invention belongs to the technical field of detection and analysis and relates to a method of on-line quickly detecting total anti-oxidizing property of samples through high performance liquid chromatography (HPLC) and a free radical detection system. The high performance liquid chromatographic analyzer includes, in a successively connected manner, a sample injector, a quaternary gradient pump, a chromatographic analytic column, a column oven and a photodiode array detector. The free radical detection system includes a free radical reaction module, a temperature controlling box, a high-pressure pump and an ultraviolet visible light detector. The free radical reaction module is connected to the high performance liquid chromatography. The temperature controlling box is used for controlling the temperature of the free radical reaction module. The high-pressure pump is connected to the free radical reaction module. The free radical reaction module is connected to the ultraviolet visible light detector. A C18 reversed-phase chromatographic column is employed as a chromatographic analytic column and is 20*3.9 mm and 5 [mu]m in model. The free radical belongs to chemical free radicals. The method is complete and beautiful in peak pattern without trailing phenomenon, is accurate in test result, is quick, is high in selectivity and is high in sensitivity.

Description

technical field [0001] The invention belongs to the technical field of detection and analysis, and relates to a method for quickly detecting the total oxidation resistance of a sample online by using high-performance liquid chromatography and a free radical detection system. Background technique [0002] Since the 1980s, due to the safety considerations of synthetic antioxidants, people tend to develop and use natural antioxidants more and more. Isolation of botanical components from herbs, fruits, and vegetables as preservatives in food, cosmetics, and pharmaceuticals has become a research hotspot. For example, fruits, vegetables, milk, fat, egg yolk and sea fish are all good sources of antioxidants. It has been reported that natural antioxidant substances with potential antioxidant properties include: tea polyphenols, soybean isoflavones, Lycium barbarum polysaccharides, natural astaxanthin, lutein, lipoic acid, α-linolenic acid (ALA), lycopene, Vitamin E, vitamin C, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N21/31
Inventor 周志钦于杰张华席万鹏张文林
Owner SOUTHWEST UNIVERSITY
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