GB/T 24903-2010 Inspection of grains and oils Determination of resveratrol in
peanuts High performance liquid chromatography GB/T24903-2010 standard download decompression password: www.bzxz.net This standard specifies the principle, reagents and materials, instruments and equipment, operating procedures and result calculation for the determination of resveratrol content in peanuts by high performance liquid chromatography. This standard is applicable to the determination of resveratrol content in peanut fruit and peanut kernel. The detection limit of resveratrol in the sample is 0.1MG/KG. ||
tt|| Appendix A of this standard is an informative appendix.
This standard is proposed by the State Administration of Grain. || tt||
This standard is under the jurisdiction of the National Technical Committee for Grain and Oil Standardization.
The drafting units of this standard: Nanjing University of Finance and Economics, Jiangnan University.
The main drafters of this standard: Wang Haifeng, Yuan Jian, Yang Xiaorong, Zhang Lianfu.
The clauses in the following documents become the clauses of this standard through reference in this standard. For all dated referenced documents, all subsequent amendments (excluding errata) or revisions are not applicable to this standard. However, parties that reach an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For all undated referenced documents, the latest versions are applicable to this standard.
GB/T6682 Specifications and test methods for water used in analytical laboratories

ICS 67.200.20
National Standard of the People's Republic of China
GB/T24903—2010
Inspection of grain and oils-Determination of resveratrol in peanut
High performance liquid chromatography
Inspection of grain and oils-Determination of resveratrol peanut by high performance liquid chromatography Issued on 2010-06-30
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China Administration of Standardization of the People's Republic of China
Implementation on 2011-01-01
Appendix A of this standard is an informative appendix.
This standard is proposed by the State Administration of Grain.
This standard is under the jurisdiction of the National Technical Committee for Grain and Oil Standardization. The drafting units of this standard are Nanjing University of Finance and Economics and Jiangnan University.
The main drafters of this standard are Wang Haifeng, Yuan Jian, Yang Xiaorong and Zhang Lianfu. GB/T24903—2010
1 Scope
Grain and oil inspection Determination of resveratrol in peanuts High performance liquid chromatography
GB/T24903—2010
This standard specifies the principle, reagents and materials, instruments and equipment, operating procedures and result calculation for the determination of resveratrol content in peanuts by high performance liquid chromatography.
This standard is applicable to the determination of resveratrol content in peanut fruit and peanut kernel. The detection limit of resveratrol in the sample is 0.1mg/kg. 2 Normative references
The clauses in the following documents become the clauses of this standard through reference in this standard. For any dated referenced document, all subsequent amendments (excluding errata or revised versions) are not applicable to this standard. However, parties to an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For any undated referenced document, the latest version shall apply to this standard. GB/T6682 Specifications and test methods for water for analytical laboratories 3 Principle
Resveratrol in the sample is extracted with ethanol-water solution. After centrifugation of the extract, the supernatant is taken and measured by high performance liquid chromatography equipped with an ultraviolet detector, and quantified by the external standard method. 4 Reagents and materials
Unless otherwise specified, all reagents used are analytical grade, and the experimental water should meet the secondary requirements in GB/T6682. 4.1 Anhydrous ethanol.
4.2 Methanol: chromatographic grade.||t t||4.3 Acetonitrile: chromatographic grade.
4.4 Glacial acetic acid.
4.5 85% ethanol solution: Take 850 mL of ethanol (4.1), add 150 mL of water, and mix. 4.6 Liquid mobile phase: acetonitrile + water + glacial acetic acid = 25 + 75 + 0.09. Take 250 mL of acetonitrile (4.3), add 750 mL of water and 0.9 mL of glacial acetic acid (4.4), mix, pass through a 0.2 μm filter membrane (5.6) and degas. 4.7 Resveratrol standard: purity ≥ 99%. 4.8 Resveratrol standard stock solution: Accurately weigh 12.5 mg (accurate to 0.0001 g) of resveratrol standard (4.7), dissolve it in methanol (4.2) and make up to 250 mL to obtain a 50 mg/L resveratrol standard stock solution, and store it in a dark place at 4 ℃ refrigerator for later use. 4.9 Resveratrol standard working solution: Accurately pipette 1mL, 2mL, 4mL, 6mL, 8mL, and 10mL of resveratrol standard stock solution (4.8), dilute with methanol (4.2) and make up to 50mL to obtain a series of standard working solutions (mass concentrations are 1mg/L, 2mg/L, 4mg/L, 6mg/L, 8mg/L, and 10mg/L, respectively). 5 Instruments and equipment
5.1 High performance liquid chromatograph: with UV detector. 5.2 Pulverizer: high-speed universal pulverizer, with a rotation speed of 24000r/min, or equivalent equipment. Desktop centrifuge: not less than 5000r/min, or equivalent equipment. 5.3
GB/T24903—2010
Microinjector: 10μL. wwW.bzxz.Net
5.5 Balance: sensitivity 0.01g, 0.0001g. 5.6 Filter membrane: pore size 0.2μm, diameter 25mm polysulfone membrane or equivalent. 6 Operation steps
6.1 Sample preparation
Peanut fruit samples are shelled, and peanut kernel samples are directly sampled. Take about 100g of peanut kernel sample and grind it with a grinder (5.2) for 2min3min. 6.2 Extraction
Weigh about 5g of the crushed sample (accurate to 0.01g) in a 250mL stoppered conical flask, add 60mL of 85% ethanol solution (4.5), place it in an 80℃ water bath for extraction for 45min, shake it from time to time, filter it with filter paper after cooling, wash the residue with a small amount of 85% ethanol solution (4.5), filter it, combine the filtrate, and make up to 100mL. Take 1mL~2mL of filtrate, centrifuge for 5min, the centrifugal speed is not less than 5000r/min, and the supernatant after centrifugation is used for injection and determination.
6.3 Determination
6.3.1 High Performance Liquid Chromatography Reference Conditions
Chromatographic column: Cls column, 150mm×3.9mm (inner diameter), 4μm, or equivalent. Mobile phase: acetonitrile decahydrate decaglacial acetic acid (4.6). Flow rate: 0.7mL/min.
Ultraviolet detector: wavelength 306nm.
Column temperature: room temperature.
Injection volume: 10μL.
For the chromatograms of resveratrol standard and sample determination, please refer to Appendix A. 6.3.2 Quantification
Use a microinjector (5.4) to draw equal volumes of the resveratrol standard working solution and the supernatant after centrifugation of the sample for analysis, and determine the response value (peak height or peak area). Draw a standard curve based on the concentration of the standard working solution and the corresponding peak area. Use the peak area of ​​resveratrol in the sample solution to check the standard curve and obtain the corresponding resveratrol concentration (c,). 7
Result calculation
The content of resveratrol in peanut kernel sample is calculated according to formula (1): XI=GXAXV
Wherein:
mass fraction of resveratrol in sample, in milligrams per kilogram (mg/kg); V
final volume of sample, in milliliters (mL); A—peak area value of resveratrol in sample solution; concentration of resveratrol in standard solution, in milligrams per liter (mg/L); c,
A,-—peak area value of resveratrol in standard solution; m—--mass of sample weighed, in grams (g). The determination result is calculated as the content of resveratrol in peanut kernel, and is retained to one decimal place. The absolute difference between two independent determination results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean. (1)
Appendix A
(Informative Appendix)
Chromatogram of Resveratrol
The chromatogram of resveratrol is shown in Figures A.1 and A.2. mv
Chromatogram of Resveratrol Standard
Chromatogram of Resveratrol in Sample Solution
GB/T24903—2010
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