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A detection kit for measuring small berry orac and its detection method

A detection kit and detection method technology, which can be used in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problem of lack of total antioxidant capacity of small berries.

Inactive Publication Date: 2011-12-07
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

The research on ORAC analysis methods is still in its infancy in China, and there is no standard measurement method and unified evaluation basis for the total antioxidant capacity of small berries. The present invention provides an ORAC test suitable for the determination of total antioxidant capacity of small berries The kit and its detection method use Trolox as the quantitative standard, and use the internationally accepted ORAC value to evaluate the total antioxidant capacity of small berries, which is convenient for popularization and application

Method used

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  • A detection kit for measuring small berry orac and its detection method

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Embodiment 1

[0020] Weigh 5.062 grams of fresh strawberries, add 80% methanol solution, and shake and extract at 28°C for 24 hours to obtain the sample solution. Take 5 microliters of sample solution and dilute to 1 ml with phosphate buffer, which is a 200-fold dilution. Dilute into 400, 800, 1600 times dilutions respectively according to the double dilution method. Add 100 microliters of sample solution (200, 400, 800, 1600 times dilution) on the 96-well microtiter plate, and then add 50 microliters of 0.4 micromol fluorescein per liter with a twelve-channel multi-channel pipette Mix the sodium reagent and react for 15 minutes at 37°C. Use a twelve-channel multi-channel pipette to add 50 microliters of 60 millimoles per liter of AAPH solution, and immediately measure the fluorescence intensity with a fluorescent microplate analyzer. The processing steps for the Trolox standard solution are the same as above, except that the dilution concentrations of the standard are 0, 2, 4, 8, 12, and 1...

Embodiment 2

[0022] Weigh 5.113 grams of fresh blueberries, add 70% ethanol solution, and shake and extract at 28°C for 24 hours to prepare a sample solution. Take 5 microliters of sample solution and dilute to 1 ml with phosphate buffer, which is a 200-fold dilution. Dilute it into 400, 800, 1600, 3200, 6400 times dilutions according to the double dilution method. Choose 800, 1600, 3200, 6400 times dilutions as the sample solution, add 100 microliters of sample solution to the 96-well microtiter plate, and then add 50 microliters of fluorescein sodium reagent with a twelve-channel multi-channel pipette After mixing and reacting at 37°C for 15 minutes, add 50 microliters of AAPH solution with a twelve-channel multi-channel pipette, and immediately measure the fluorescence intensity. The processing steps for the Trolox standard solution are the same as above, except that the dilution concentrations of the standard are 0, 2, 4, 8, 12, and 16 micromolar per liter.

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Abstract

The invention relates to a method for measuring the total antioxidant capacity of small berries, provides an ORAC detection kit suitable for the determination of the total antioxidant capacity of small berries and a detection method thereof, and belongs to the technical field of activity detection and determination. The kit includes: antioxidant standard substance Trolox storage reagent, fluorescent substance fluorescein sodium storage reagent, free radical generator AAPH reagent, phosphate buffer, and fluorescent microwell reaction plate, which is made of a plastic or hard paper material Boxes are packed together to make up. The ORAC detection method for the total antioxidant capacity of small berries comprises the following steps: small berry sample pretreatment; detection with reagent kits; and analysis of detection results. By mixing the sample and the reagent in a certain volume ratio, the fluorescence microplate analyzer is used to detect the change of the fluorescence intensity of the complex, thereby measuring and calculating the total antioxidant capacity of the small berry. The invention can provide a complete set of total antioxidant capacity test of small berries, has high sensitivity and good accuracy, and is convenient for popularization and application.

Description

1. Technical Field [0001] The present invention is a method for determining the total antioxidant capacity of small berries, provides an ORAC detection kit and a detection method suitable for determining the total antioxidant capacity of small berries, and belongs to the technical field of activity testing and determination. 2. Background technology [0002] Small berries, including strawberries, blueberries, raspberries, blackberries, etc., are popular all over the world with their unique flavors and nutritional and health value, and are "super fruits". Small berries are rich in polyphenolic active substances such as vitamins and anthocyanins, which have the function of anti-oxidation and scavenging oxygen free radicals. They are the third-generation health-care fruit that integrates nutrition and health care. Studies have shown that aging and various diseases are related to oxygen free radicals. Therefore, foods that are closely related to health, including the determination of...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 黄午阳李春阳刘文旭闫征王乃富
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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