Some standard content:
FCS67.040
National Standard of the People's Republic of China
GB/T5009.149—2003
G/17335.--1998
Determination of chroma in foods
Determination of chroma in foods2003-08-11Promulgated
Ministry of Health of the People's Republic of China
National Administration of Standardization
Implementation on 2004-01-01
This standard replaces GB/T17535—1998 Determination of chroma in foods. (H/T5009.149—2003
Part 4 Chemical analysis methods 3 This standard has been revised in accordance with GR/T23001.4—2001-Standard Writing Rules 2.
This standard is proposed and approved by the Ministry of Health of the People's Republic of China. The responsible drafting unit of this standard is the Institute of Labor Hygiene and Occupational Diseases, Chinese Academy of Preventive Medicine, and the participating units are the Food Safety Supervision and Inspection Institute, Ministry of Health.
The founders of this standard are Feng Yanpiao, Tu Mei and Yang Zuying. The original standard was first revised and issued in 1999, and this is the first revision.253
G1/5009.1492003
Cabinet yellow as a coloring agent has been listed in GB2760--1SS6 Hygiene Standard for Food Coagulants, with a maximum use amount of 0,=/kg 5
1 Scope
Determination of yellowing agent in food
This standard specifies the high performance liquid chromatography method and high performance liquid chromatography method for the determination of yellowing agent in a wide range of foods. This standard is applicable to the determination of yellowing agent in beverages, wines and cakes. The detection limit of the first method is a.?u/ml: the standard range is mg/ml~22g/ml. The first method is high performance liquid chromatography method
2 Principle
GB/T 5009. 149—2003
After the yellow in the sample was extracted and chemically treated, it was determined by high performance chromatography, and the main components of yellow were determined by the time and distance of the sample, as the reference substance.
3 The reagents were pure, and the water was algae water
3.1 Methyl acetate
3.2 Petroleum aldehyde 65~.
3.4 Ditert-butyl ether,
3.5 Main yellow pigment
3. The yellow pigment was obtained. || tt||3.7 Substance test solution: weigh 2.75 mg of low standard substance, dissolve it with methanol, and dilute it with 100 ml of methanol to obtain 27.5 g/mL constant temperature.
3.8 Substance standard working solution: pipette 0.2, 0.4, 6.0, and 0.1 mT of substance standard solution into 10 mL volumetric flasks respectively, add 100 ml of standard solution to make up to 0.1 mT in sequence to obtain substance test standard series of 3, 1.5, 11.1, 13.5, and 22.1 g/mL. 4.1 Small powder machine.
4.2 Water-soluble.
4.3 High performance liquid chromatography system: Waler's M510 pump, L'6K feeder, island pool RF-535. Fluorescence detector, Bluchip/PC meter and Baeline81C color control program, 5 analysis step BA
5.1 Sample treatment
5.1.1 The sample is cooked, stirred and the dioxide cap is removed Or super total gas, after oblique hook. Pass through microporous membrane (., Am filter, the filtrate is prepared for HPI analysis.
5.1.2 Alcohol; the sample is filtered through microporous cell iodine and prepared for HPLC analysis. 5.1.3 Black spot Take 1ng sample and put it into 1U0m medical plate, heat it with 50mL petroleum aldehyde and flow it for 1min, put it into a wet sand core funnel, wash the residue with alcohol 5 times, add the washing liquid to the decompression and concentrate the petroleum fermentation extract, and put the residue into the fume hood until there is no stone carving smell. Extract with medium for 3 to 5 times, 30 ml each time, until the extracted solution has no yellow color. Filter with a temperature of 100 °C for 20 seconds, pass the extracted solution through a 29
GB/F5009.149—2003
microporous filter membrane, filter and store in a refrigerator. 5.2 Determination
5. 2. 1 HPLC reference conditions
Chromatographic column: particle size 5 um ()DS Cx 150 mm×1. t mm: mobile phase: methanol-water (35-55)
flow rate: 0.3 mL/min
wavelength, 240 m,
5.2.2 Standard curve
Under the experimental conditions, the standard test solution, 2, 4, T, were injected respectively for ITL analysis, and then the standard curve was drawn using the height against the concentration of the test solution.
5.2.3 Sample determination
Under the experimental conditions, add 3 μT. 5.: The sample is dissolved in water and analyzed by HPLC. The peak is compared with the standard peak to determine the content of the test substance.
6 Calculation of results
According to the following calculation:
×1000
Content of yellow pigment in the sample, in grams per kilogram (g/kg); A—the total volume of the sample in the injection, in micrograms per milliliter (μg/mL); V—the volume of sample preparation, in microliters (μL); m—the mass of the sample, in grams (g).
7 Precision mouse
The absolute difference of two independent determination results obtained under repeatability conditions shall not exceed 5%. The second method is thin and thick chromatography.www.bzxz.net
8 Principle
The yellow pigment in the sample is extracted with an organic solvent and purified to remove the lower substances. After the sample is spotted and developed, it will appear as a black spot under the TIV254HT lamp. It is compared with the standard for qualitative and approximate quantitative analysis. 9 Reagents
All reagents used are analytically pure, mainly distilled water, 9.1 methanol.
9. 27 alcohol.
Ethyl acetate.
Two ketones.
Trimethylmethane,
Silica gel core F251. Developing solvent for thin layer chromatography: ethyl acetate + acetone + methanol + water (5+5-111). 9.8
9. 9 Developing agent, trifluoromethane + alcohol (6 + 3). 10 Only part
10.1 Whole pond concentrator.
10.2 Layer plate coater.
10.3 Glass plate, 1 cmx20 cm.25 enX20 cmm10.4 Chromatography.
10.5 UV24nm fluorescent lamp.
10.6 Micro syringe.
Operation method
11.1 Sample treatment
TLC test is performed on the samples of alcohol, alcohol and egg after treatment, and the results are shown in 11.2.2. 11.1.1 Alcohol: Collect 100mL of the sample, concentrate under reduced pressure until there is no alcohol smell, then extract with ethyl acetate, 30mL each time, extract 3 to 5 times, until there is no yellow color, then combine the liquid, take the bottle, and concentrate under reduced pressure until there is no yellow color. About 20mL is left, which is used for urine analysis.
11.1.2 Drinks: Take 100mL of the sample and extract with ethyl acetate, 50mL each time, extract 3 to 5 times, until there is no yellow color, then combine the liquid, concentrate under reduced pressure until there is no alcohol smell, and 20mL is left, which is used for urine analysis. 11.1.3 Cake: Take 1 μg of the powdered sample, add a little seaweed, mix well, blow dry the sample with hot air (for modeling), add 50 ml petroleum ether, stir, let stand for a while, discard petroleum ether, repeat this process three times to remove fat, grind after each use, transfer to a Soxhlet extractor, use methyl methacrylate to extract the pigment until no yellow surface is left. When all the pigment is extracted, concentrate to about 100 μl. This liquid is reserved for layer chromatography. 11.2 Determination 11.2.1 Spotting: Take a commercially available silica gel GF354 fluorescent plate, spot 0.5 μL of the sample at 70° above the bottom edge of the plate, and spot the plate with a 2L color standard.
11.2.2 Expand, and place the sample and standard plate already spotted in 11,2.1. Use 5.8,3.9 to spread and hold until the yellow is clearly visible and then collect and dry. Compare with the standard spot: insert the yellow R, the value is 0.64 and L.50. The yellow spot is 0.11, _5. The sample and the standard are consistent, which proves that the silk in the sample is yellow. Table 261
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