GB/T 5009.23-2003 Determination of aflatoxins Bl, B2, G1 and G2 in foods
Some standard content:
ICS67.040
National Standard of the People's Republic of China
GB/T 5009.23--2003
Replacement #GB/5909.231S96
Determination of aflatoxins B, B2, G1, G2 in foods2003-08-11Promulgated
Ministry of Health of the People's Republic of China
Standardization Administration of the People's Republic of China
Implementation on 2004-01-01
GB/T 5009.23--2003
This standard replaces GB/T 5909.2-1996 Determination of aflatoxins B, B2, G1, G2 in foods. The main modifications of this standard compared with GB/T 5909.23-16 are as follows: The Chinese name of the standard has been changed to \Determination of Yellow Oxygen in Dangerous Goods - Part 1: Chemical Analysis Methods in accordance with GB/20001.4-2CD Standard Preparation Rules. The structure of the standard has been modified.
This standard was drafted and managed by the Ministry of Health of the People's Republic of China. The first method of this standard was drafted by the Institute of Nutrition and Food Production, Chinese Academy of Preventive Medicine, and Qingdao Import and Export Commodity Inspection and Quarantine of the People's Republic of China.
The second method of this standard was drafted by the Institute of Nutrition and Food Production, Chinese Academy of Preventive Medicine, Beijing Municipal Institute of Food Safety and Health, Chinese Academy of Preventive Medicine, and the Ministry of Health's Food Safety Supervision and Inspection Institute.
This standard was first issued in 1985 and revised for the first time in 1999. This is the first revision. 19
1 Scope
Determination of aromatic benzoate BG in food
This standard specifies the determination method of aromatic benzoate C in various foods. This standard is applicable to the determination of aromatic benzoate G in food. GB/T5009.23—2003
The amount of aromatic benzoate BG on the low grid plate is 0.0. The detection limit of this method is 0.0:3:1/g. The detection limit of this method is 0.5:1/g.
2 Normative references
The following clauses are applicable to this standard and become its clauses through the introduction of this standard. Any document with a date and its subsequent amendments (excluding the amendments) are not applicable to this standard. However, the parties who have not reached an agreement on the standard shall study whether the latest versions of these sections can be used. For any referenced document without a date, its latest version is applicable to this standard. GB/T5003.222003 Yellow Industry Recommended B, First Method for Medical Chromatography
3 Principle| |tt||After extraction, concentration and thin layer separation, under 6Fnm external light, yellow poison B, B: produces blue-purple light, yellow poison G produces yellow colorless fluorescence, which is quantified according to the luminescence displayed on the explosion layer, 4 reagents
4,1 Same as CBT5005.22-13 3.1-~3.13,4.2 Sodium hypochlorite solution (for consumption): Preparation method see (B/T5303.222003 3.16.4.3) - ethanol - water (43+35+19) developer: Collect this ratio The drop was placed in a concentration stagnation bucket, shaken for 5 minutes, and left to stand overnight. The upper and lower layers of the drop were stored in bottles respectively. Do not accidentally mix the droplets. If the liquid is mixed, add water to it. After it becomes clear, stop heating, take the upper layer of solution for development, take a certain amount of the lower layer of solution, and put it in the development medium. Put the thin layer plate in the receiving medium, and develop it after pre-saturation with 4.4 tin sulfate 1.-3)
4,5 Huangshan belt each high B.,B,,G1,G: Standard solution is as follows: 4.5.1 Single standard Liquid (10g/ml.): weigh aflatoxin BG, standard product 1mg~1.2mg: dispersed aflatoxin B., G standard product 0.5--0.6g·monthly extract ethyl ester mixed solution as the reagent. Preparation method, concentration and determination of concentration Record 1:15009.22233313.14
The content of aflatoxin HR, G., and the maximum absorption filter length and elimination coefficient of the aflatoxin are shown in Table 1 Table 1
Maximum absorption peak wavelength/nm
Extinction coefficient
12 BC9
20 963
4.5.2 Service Standards: The following standards are used for the mixed reduction of acetylene and chlorine. GH/T5039.29-2003
4.5.2.1 Yellow standard is used for 11% yellow GC. 0.2g yellow, 2 for fast use.
4.5.2.2 Cost-aided penetration rate tolerance standard: 6% yellow per liter, 002% yellow zero storage requirement Ts. for low detection.
5 Instruments
GB/T 5u3.22-20s4.1~.4.U
6 Analysis steps
6.1 Sampling
same, (/ 503. 33-2(05 in 5. . 6.2 Extraction
GB/T5309.22
20035.2.
6.3 Determination
6.3.1 One-way development method
6.3.1.1 Preparation of temperature-controlled absorption
(B/15309.2220035.3.1.1.
6.3.1.2 Spotting
Same as GE/T5:13.22--2003 in 5.3.1.2. The dropping pattern is as follows. First point: 1Gα1. Huangqu needs two series mud containing standard working solution II. Second point: 2CμL sample,
First 3.0. Sample 10% effective rate of the mixed use of the fourth point, 2UL sample hungry + 1UL yellow koji poison mud standard use of wave 1. G.3.1.3 Development and observation
yellow koji BBG ratio of the transfer value is arranged in order of B>B>G>G Add 1cm of anhydrous ethyl alcohol in the development mold, pre-expand 12cm, take out the machine. Then add 10mL of acetonitrile-dihydrochloric acid (8 + 93) in the development mold, expand 1Vcm~12cm, and take out. Result: The method is as follows. E 3.1.3.1 By adding aflatoxin to the sample point and then using the standard 1 change, the fluorescence points of aflatoxin B, 3, and GG can be overlapped with the fluorescence points of aflatoxin B, 3, and GG in the sample. If the sample is negative, the third point on the sample layer is 0.4, 0.2, 0.01, 0.03, and 0.10, respectively. It can be used to check the presence of aflatoxin B in the body. If the detection point is normal, it can be classified as a ratio, then the positioning effect is achieved. According to the yellow curve of the first point on the pole, i, G:, G, according to 0.002, 3.01, (.002, 0.002, Co: E. ±, it plays a positioning role. 6.3.1.3.2 If there is no blue-purple fluorescent spot at the second point corresponding to the yellow curve *B: B, or there is a yellow-green fluorescent spot at the corresponding position of the total curve, it means that the yellow curve of the sample contains less than 5g/kg; BG is less than 2.5/kg. If there are more than 1 fluorescent spot at the cabinet level, a confirmation test is required. 6.3.1.4 Confirmation test
6.3.1.4. 1 Huang Dian must be obtained by reacting with acetic acid to produce a derivative. It is limited to B. and G. (and blue or acetic acid do not react. B and G. will be the biological ratio of me BG1. Add two points to the edge of the thin layer plate. The first point: 101. Huangqu standard liquid
The second point: 20pL sweep liquid: bzxZ.net
Ten or more points are cut into three small drops each. Cover it. After 5 minutes, use a hair dryer to blow hot air for 2n. Make the hot air blow to the temperature of the thin layer plate not higher than: Then cut the following two points on the thin layer plate. The third point: 10 called. The first song is used again to meet the standard red liquid. 192
The fourth point: 2 mixed liquid.
GB /T5009.23-~2003
Then observe the sample under normal external light to obtain whether it produces the same biological target as aflatoxin B: or G:. The three and four points of acetic acid can be used as blank controls for the sample and the standard derivative. 6.3.1.4.2 The aflatoxin test of B: or G: is carried out by placing the sample in acetic acid (46+35+19) for development. If the standard point and the sample point coincide with each other, it can be confirmed.
6.3.1.4.3 On the developed thin plate, the matrix is multiplied by sulfuric acid (1-3) and the aflatoxin B:, B:, G: and G: all turn into yellow fluorescence. 6.3.1.5 Dilution Quantification
The temperature of the zero-toxic aflatoxin B, G and the standard temperature of the aflatoxin B, G and the standard temperature of the standard temperature are respectively 0.If the fluorescence intensity of the sample is consistent with that of the detection point, the content of BG in the test sample is 5/g:, and the amount is R/kg. If the fluorescence intensity of any yellow poison in the sample is stronger than the detection limit, it is necessary to quantify it step by step until the fluorescence intensity of the sample scattered point is consistent with the fluorescence intensity of the detection limit point. The determination and calculation method refer to 5.3.1.5 and 3.3.1.5 in G13/10C9.222003. 66.3.2 Two-way opening method
6.3.2.1 Two-point addition method
6.3.2.1.1 Take three full plates and add the standard aflatoxin solution to the plate 3m from the bottom. That is, drop 10% of the standard aflatoxin solution at 8cm-1cm from the edge of the plate. Add 20% of the standard aflatoxin solution at 2.8cm-3.0cm from the left edge. Then add 10% of the standard aflatoxin solution to the sample point of the second plate. Add 1% of the standard aflatoxin solution to the sample point of the second plate. 6.3.2.1.2 Same as 5.3.2.1.2 of GR/T 5009.222303. 6.3.2.1.3 Observe and evaluate the results:
6.3.2.1.3.1 Observe the first and second plates under ultraviolet light. If the second point of the second plate is within the range of the standard points of G, G, and G, the first and second plates shall be inspected. If the minimum detection amount appears at the corresponding place, and there is a fluorescent spot at the same position on the first plate as on the second plate, then the content of yellow soda ash B and G in the sample is below 2.6.3.2.1.3.2 Make the first report on the fluorescent spot at the same position on the second plate, then compare the first plate with the second plate to see whether the fluorescent spot at the second point on the third plate overlaps with the standard G point. If so, conduct the confirmation test according to 5.3.2.1.3.5. 6.3.2.1.3.3 Confirmation test of yellow soda ash B and G: Place a thin plate on the fourth plate, and add 10% yellow soda ash mixed with standard 1% diacetate at a distance of 0.2cm to 1cm from the second plate, 2.8cm to 3cm from the left edge.Flow into the separating funnel, blood = methane 1UmL, this sample splash 1mL is equivalent to 2.0R sample. 10.1.4 Separation, yellow: take 21 of the source sample, put it in a bottle with a seat, add 100L of alcohol water, 20m1. Swing 30in, put it in a clean piece, pass it through a folded rapid qualitative filter paper for 5nIv.I. Check the volume, collect the methanol water filtrate FL (equivalent to 1C sample, about 12g of sample contains 5L of water), put it in a separatory funnel, add 1Cml of chlorine in the flow, connect 1.1.1\lightly diffuse 2mit-mia\ and it is full! TcJ is equivalent to .0R style, 10.2 Determination
10.2.1 Preparation of column tube: Use a small amount of defatted phase as the bottom and use thick iron wire to make it sure, and then put it on the hanging tube rack. Add people in sequence with a height of J.5cm and then find 114
GB/T 5009.232003
sulfuric acid (0 day ~.1) 0cm group magnesium rate absorbent, heart.cm no rice feeling sodium (80 days-00 days), 1.5ctr neutral annual aluminum 1.r1 acidic lead oxide: 3m water breaking sodium (4) glycerol-80 days) pre-blocked with a little cover of absorbent cotton, when loading and testing, each reagent should be appropriately filled, and the surfaces of the two reagents should be flush. The column should be used as soon as it is loaded to avoid the decrease of water absorption activity in the air. 10.2.2 Microscope: In the prepared microscope, add oxygen at 1.0m above the light. At the same time, add 1m dioxane and then add yellow T3: standard push liquid (0.1, 0.50, 100L). Each of the three microscopes should be equivalent to dry yellow B. When the filter reaches the top layer, add 0, 5, and 1mL of trichloromethane (-10.9> developing agent. When developing, keep the tube vertical and keep the developing agent flowing. After the study, the result can be observed. 10.2.3 Results and determination: The results were observed under a 565nm ultraviolet lamp. After analysis, the sample tube is compared with the U, 5, and J5 emission standards. If the adsorbent layer in the sample tube is not as good as the sample (the silicon magnesium adsorbent layer in the sample shows a yellow fluorescence in the / below), it is necessary to further perform confirmation determination by thin layer chromatography, but there is no need to re-excite the sample. The operation is as follows: accurately pipette 1ml of chloroform extract (equivalent to 1R or 8P sample) from the source, and add a small amount of benzene 7. The reaction mixture (S812) is transferred into a graduated small tube several times to a sufficient volume of 1.0mL, and the volume is filled to full to reach the confirmation level of thin layer chromatography, and the content of yellow soda ash 5 is measured.
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