GB/T 5009.27-2003 Determination of benzo(a)pyrene in foods

This standard specifies the determination method of benzo(a)pyrene in food. This standard is applicable to the determination of benzo(a)pyrene in food. The detection limit of this method is 1ng/g when the sample volume is 50g and the spot sample volume is 1g. GB/T 5009.27-2003 Determination of benzo(a)pyrene in food GB/T5009.27-2003 Standard download decompression password: www.bzxz.net

[CS 67.040
National Standard of the People's Republic of China
GB/T5009.27—2003
Replaces GB/T5005.27—16
Determination of benzo (a) prrene in food
foods2003-08-11 issued
Ministry of Health of the People's Republic of China
China National Standardization Administration
2004-01-01 implementation
CB/I5009.27—2003
This standard CB/T50(9.27.-996 Determination of benzo(a) in foods\. Compared with GH/T6n09,27—1996, this standard has the following changes: the name of the standard has been changed, the Chinese name of the standard has been changed to the standard of the product, and the main part of the standard is the first part: Chemical The structure of the original standard was revised in accordance with CB/T2UJL.4 in the "Analytical Method". The standard was proposed and coordinated by the Ministry of Health of the People's Republic of China, and was drafted by the Beining Provincial Health and Epidemic Prevention Station, the Hongsu Provincial Health and Epidemic Prevention Station, the Guangxi Zhuang Autonomous Region Health and Epidemic Prevention Station, the Shanghai Health and Epidemic Prevention Station, and the Xinjiang Uygur Autonomous Region Health and Epidemic Prevention Station. This standard was issued in 1991 and revised for the first time in 1996. This is the second revision. 21 Determination of Additive () Flower in Food This standard defines the method for the detection of additives in food. This standard is for the detection of additives in foods. Method detection limit: The sample volume is 1 when the amount of the sample is 1. The first method is fluorescence spectrophotometry. 2 Principles GB/T5009.27-2003. The organic flower is taken from the filter paper, or after saponification, the organic makeup liquid is purified by liquid-liquid separation, and then separated on acetylated filter paper. The solidified rice (the rice) is irradiated with blue-purple light. The paper part with the flower after separation is cut off, and after being immersed in the mountain, the light flux is measured with a spectrophotometer and the cup is covered with a fixed cover. 3 Reagents 3.1 Distilled rice ether (or right oil ether, boiling point 6~) heavy benzene is treated with alumina to obtain light. 3.3 1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1-hydroxy-2-hydroxy-2-hydroxy-1 3.12 Cooled filter paper: Cut the filter paper for rapid chromatography into strips of 30cm×4cm, and put the strips into a 500ml beaker filled with ethyl anhydride (18ml, 130ml acetic anhydride, 0.1ml aldehyde). Drain the strips thoroughly and keep the solution temperature at 21°C or above, stirring for 2 hours. Leave overnight, take out the strips, put them in a ventilated cabinet, and place them in a large Yong7 bath for 4 hours. After aspiration, press them flat on a porcelain plate covered with half filter paper under indoor wind. 15 to 18 strips can be processed at a time. 3.13 Standard solution of polyvinyl alcohol (m-butyl alcohol): 3ml brown wax solution (10.0-1% benzophenone) and the volume should be increased by 5 times. Place in ice and keep in 3.14ml standard solution. Pipette 1.00 ml of standard solution into a 1.00 ml volumetric flask and dilute with the same method to the same concentration. Then add 1.0 and 0.1 ml of standard solution per liter. Store in refrigerator.
4 Collector
4.1 Collector.
GH/T 5009.27—2003
4.2 Chromatographic needle with inner diameter of 10mm and length of 5mm: with inner diameter funnel of 2mm on the upper end and inner diameter of 50m~10m on the lower end, and with a flexible case on the lower end.
4.3 Chromatographic reagent (commercial)
4.4 KD·glass gradienter.
Ultraviolet lamp: with wavelength nm or 25m oil film 4.6 Reflux chemical, condenser tube connected to the flask. Tissue cotton shredder:
4. Daylight fluorescence spectrometer.
5 Analysis steps
5.1 Sampling extraction
5.1.1 Food with low moisture content: Weigh 40.0g--60.0g of pulverized sample, put it into a container, and moisten the sample with 70mL. Hexane. Put E~g of oxide, 100mL of ethanol (95%) and 50mL~801mL of cyclohexane in the container: then connect the skin retardant, reflux extraction on 9 water for 6b-8h, heat the saponified solution into 300mL, store in a bucket, and pour the cyclohexane in the filter paper into the filter paper. 5%), divide into two receiving bottles, combine the wash and dry funnel: add 10mL water, add charcoal for 3min, let stand for separation (about 20min), the lower layer is added to the second separating funnel, and then extracted with 71mL cyclohexane. After stratification, discard the lower layer, combine the calcined layer in the first separating funnel, and rinse the second separating funnel with 6rrl.31ml. cyclohexane, combine the washes, wash with water, combine and extract with cyclohexane once, 100mL each time, and combine the three washes in the original second separating funnel, extract twice with cyclohexane, 30ml each time , shake for 0.5min, discard the water layer after stratification, collect the cyclohexane layer and put it into the first separatory funnel, add 5mL ~ 50mL water, concentrate under reduced pressure to 4mL, add appropriate amount of anhydrous sodium sulfate for dehydration, 5.1.2 Vegetable oil: weigh 30.1g ~ 2:.C of mixed oil sample, wash with 19mL cyclohexane in batches into 2mL separatory funnel, extract once with cyclohexane and saturated monomethyl methacrylate, 40mL each time, shake 1mL, combine the dimethyl methacrylate extracts, extract with 40mL cyclohexane saturated with dimethyl formamide: once, discard the cyclohexane layer: dimethyl formamide The extracts were combined in a 500-mL separatory tube containing 245 mL of sodium thiocyanate (2 g/L), mixed, and allowed to stand for a few minutes. The cyclohexane extracts were extracted twice with 191 mL each time and incubated for 3 min. The cyclohexane extracts were combined in the first 500-mL separatory tube. Dimethylformamide may also be used instead of dimethylformamide. The cyclohexane extracts were extracted twice with 100 mL each time using 4°C-50% water, and the extracts were extracted for 0.5 min. The aqueous layer was discarded and the hexane layer was collected and concentrated to 41 mL under reduced pressure in a 50°C ~ 6% water bath, and dehydrated with anhydrous sodium thiosulfate. 5.1.3 Fish, fish and their products: Weigh 50.nR~50.0R of the mixed sample, stir with dead water sodium sulfate (the ratio of sample to dead water sodium sulfate is 11 or 12. If there is too much water, the sample needs to be dried at 60°C first), put it into the filter paper, and then connect the fat-assisted extractor. Add 190% cyclohexane and extract at 0℃ for 6h. Then put the net extract into a 250mL separatory funnel, and then add 6mL~Bm Before washing the filter paper with n-cyclohexane, wash and combine in a 25 cm3 separatory funnel and extract three times with dimethylformamide saturated with cyclohexane. Continue according to the method. 5.1.4 Vegetables: Weigh 100.0 g of clean and air-dried edible vegetables, chop them into small pieces and put them into a tissue extractor with 150 ml of acetone. Heat for 2 min. Add a little deionized water to the small funnel and add 100 ml of acetone to the separatory funnel. The residue was separated and washed with L-inone. The washing liquid and the filter were placed on a filter plate. 100 mL of water and 1 ((m) cyclohexane were added to separate and extract for 30 min. The layers were allowed to stand and separated. The cyclohexane layer was transferred to another 50 mL separating funnel. The water layer was extracted twice with 1 CCml of cyclohexane. The cyclohexane extracts were combined and put into the first separating funnel, and then separated and washed with 20 mL of water for 2 times. The cyclohexane was collected and concentrated to 2 mL under reduced pressure in a 50-60 °C water bath. Add Dehydrate with anhydrous sodium sulfate.
5.1.5 Beverages <If the carbon dioxide is removed by heating in a water bath>: Pipette 5.ml~00.0ml. Place the test rod under 550ml. Dispense into a funnel, add 2g of activating sodium hydroxide to dissolve, add 50ml of cyclohexane and shake for 1min, separate the layers, and separate the water into the second dispenser. Then collect once with 50ml of cyclohexane, combine the collected hexane solutions, and add 103ml each time.Vibrate with water, wash twice, collect cyclohexane and concentrate to 25\L under reduced pressure on 5u%~6(water lead), add appropriate amount of sodium sulfate and water. GB/T5009.27—2003
5.1.6 Pastry, take 50.0%~60.C ground and filter paper, and operate according to 5.1.: Use 70)I. Cyclohexane to wet the sample... ". In 5.1, 1, E.1.3-5, 1., each operation, can be replaced by petroleum aldehyde, but with petroleum ether extraction chain to dry, make 35mL cyclohexane float.
5.2 Purification
5.2.1 Dry the lower end of the chromatographic column filled with natural glass, and then filled with 5T-6cm Calcium oxide, lightly cover the surface to fill the chemical layer, especially the hollow, level the top surface, then add 5cm1~6cm crushed adsorbent, and then add 5cm~-5cm anhydrous sodium aldehyde, elute the selected chromatography column with 31uL of hexane, close the piston when the hexane is lowered to the anhydrous sulfuric acid layer, 5.2.2 put the sample Y ring extracted into the chromatography column, set the temperature, and the throttling is recommended to be 1mL per minute, If necessary, appropriate force can be used to remove the cycloalkane liquid until it drops to the anhydrous sodium sulfide layer, and then the cycloalkane liquid should be observed under ultraviolet light until the blue fluorescent substance completely descends from the alumina layer. If the volume is too low, appropriate amount of liquid can be added, and the liquid can be collected and concentrated on a water bath under reduced pressure to 50-0.1 mmol/l. It can be determined according to the content of the raw material in the sample. The blood should not be evaporated. 5.3 Separation
5.3.1 Cartridge 7. At a 5c mark on the filter paper strip, draw a line with a pen as the starting line, collect a certain amount of the concentrated liquid after standardization, and spot it on the paper strip. Blow the air from the back of the strip and select the reagent. The same strip should be spotted with the standard solution (1=/mT). When spotting, the vertical grid of the spot should not exceed 31mrr. The chromatographic (simple) contains a developing agent, and the lower end of the filter paper strip is set with about 1% developing agent. cm, and take it out when the slide of the agent is about 2 cm. 5.3.2 Observe the developed paper strip under 365nm or 254nm ultraviolet light. Use a pen to mark the blue spots of the standard benzo(a) and the sample with the same weight: cut out these spots and put them into small colorimetric tubes, add 4mL of each, and put them into a 50--S0 water bath and shake them from time to time for 1.min.
5.4.1 Put the extracted samples and standard spots into the right cup of the fluorescence spectrophotometer In the process, 5nm is used as the excitation wavelength, and 36Sn~6Um wavelength is used for carbon scanning. The obtained spectrum is compared with the standard spectrum with the standard spectrum of () meters for qualitative analysis. 5.4.2 At the same time as the above sample analysis, a blank reagent is made, including all the reagents used for processing or sample. Read the fluorescence intensity of the sample, standard and test blank at 406m, (46+5m, (06-5)m respectively. The number calculated by adding 11 to the baseline is the calculated fluorescence intensity,
F -- Fur (FI Fu?/2
5.5ResultsThe oxygen meter
The content of oxygen in the sample is calculated by formula (2), X=[5/FXtF,-F>×1000_/(mXVVt
Wherein:
Y——The content of oxygen in the sample, in micrograms per gram (/kR): S
The mass of the standard spot of oxygen, in micrograms (ug): The fluorescence intensity of the standard spot, in millimeters (mm): The sample spot outflow concentration, in meters (mm); F
The diameter of the immersion liquid, in millimeters (mm); The volume of the sample shrinkage liquid, in liters (); The sample volume is in liters (mI.); The sample volume is in grams (g).
Calculation results are expressed to the decimal place
5.5 Precision
The difference between the two independent determination results obtained under the reproducible strip shall not exceed 2% of the arithmetic mean (2)
GB/T5009.27—2003
6 Principle
The second method is colorimetric method
The sample is adsorbed and purified on the acetic acid filter paper. The (a) standard spot with a high concentration of 100 μm is observed under a 35nm ultraviolet lamp and compared with the standard spot. 7 Reagents
F 3. 1--3. 11.
Interval 4.1--4.8.
9 Analysis steps
9.1 Extraction
Follow the method of step 5.2.
9.2 Purification
Follow the method of step 5.2.
9.3 Adjustment
Collect 5, 10, 15, 20 or 6C sample concentrate according to the test millet and (a) content and determine the 1C23 content (u) standard (0.1K/WL): Melt on the same filter, expand the filter in step 5.3.1, and take out the sample. Under the light of the external lamp, compare the intensity of the sample. When the sample contains high concentration of benzoic acid, it can be reduced and then concentrated. Try to put the test rod between the test points: 3.4 Calculation of the sample can be calculated according to formula (3). x(m1 5)/(m×V./V.)
Wherein:
x ​​is the total amount of oxygen extracted in the sample, in micrograms (g/kg)—mass of the sample extracted with oxygen, in grams (g): 1, or the total concentrated content of the sample, in liters; V.—specific volume, in liters (ml.); 22
sample weight, in grams (g).-F>×1000_/(mXVVt
In the formula:
Y——the content of extract in the sample, in micrograms per gram (/kR); S
a) the mass of the standard spot, in micrograms (ug); the fluorescence intensity of the standard spot, in millimeters (mm); the sample spot outflow concentration, in millimeters (mm); F
records the surface area of ​​the immersion liquid, in millimeters (mm); the volume of the sample shrinkage liquid, in liters (liters); the sample volume is in liters (mI).
The formula is detailed in the mass disk, in grams (g).
Calculation results are expressed to the decimal place
5.5 Precision
The difference between the two independent determination results obtained under the reproducible strip shall not exceed 2% of the arithmetic mean (2)
GB/T5009.27—2003www.bzxz.net
6 Principle
The second method is colorimetric method
The sample is adsorbed and purified on the acetic acid filter paper. The (a) standard spot with a high concentration of 100 μm is observed under a 35nm ultraviolet lamp and compared with the standard spot. 7 Reagents
F 3. 1--3. 11.
Interval 4.1--4.8.
9 Analysis steps
9.1 Extraction
Follow the method of step 5.2.
9.2 Purification
Follow the method of step 5.2.
9.3 Adjustment
Collect 5, 10, 15, 20 or 6C sample concentrate according to the test millet and (a) content and determine the 1C23 content (u) standard (0.1K/WL): Melt on the same filter, expand the filter in step 5.3.1, and take out the sample. Under the light of the external lamp, compare the intensity of the sample. When the sample contains high concentration of benzoic acid, it can be reduced and then concentrated. Try to put the test rod between the test points: 3.4 Calculation of the sample can be calculated according to formula (3). x(m1 5)/(m×V./V.)
Wherein:
x ​​is the total amount of oxygen extracted in the sample, in micrograms (g/kg)—mass of the sample extracted with oxygen, in grams (g): 1, or the total concentrated content of the sample, in liters; V.—specific volume, in liters (ml.); 22
sample weight, in grams (g).-F>×1000_/(mXVVt
In the formula:
Y——the content of extract in the sample, in micrograms per gram (/kR); S
a) the mass of the standard spot, in micrograms (ug); the fluorescence intensity of the standard spot, in millimeters (mm); the sample spot outflow concentration, in millimeters (mm); F
records the surface area of ​​the immersion liquid, in millimeters (mm); the volume of the sample shrinkage liquid, in liters (liters); the sample volume is in liters (mI).
The formula is detailed in the mass disk, in grams (g).
Calculation results are expressed to the decimal place
5.5 Precision
The difference between the two independent determination results obtained under the reproducible strip shall not exceed 2% of the arithmetic mean (2)
GB/T5009.27—2003
6 Principle
The second method is colorimetric method
The sample is adsorbed and purified on the acetic acid filter paper. The (a) standard spot with a high concentration of 100 μm is observed under a 35nm ultraviolet lamp and compared with the standard spot. 7 Reagents
F 3. 1--3. 11.
Interval 4.1--4.8.
9 Analysis steps
9.1 Extraction
Follow the method of step 5.2.
9.2 Purification
Follow the method of step 5.2.
9.3 Adjustment
Collect 5, 10, 15, 20 or 6C sample concentrate according to the test millet and (a) content and determine the 1C23 content (u) standard (0.1K/WL): Melt on the same filter, expand the filter in step 5.3.1, and take out the sample. Under the light of the external lamp, compare the intensity of the sample. When the sample contains high concentration of benzoic acid, it can be reduced and then concentrated. Try to put the test rod between the test points: 3.4 Calculation of the sample can be calculated according to formula (3). x(m1 5)/(m×V./V.)
Wherein:
x ​​is the total amount of oxygen extracted in the sample, in micrograms (g/kg)—mass of the sample extracted with oxygen, in grams (g): 1, or the total concentrated content of the sample, in liters; V.—specific volume, in liters (ml.); 22
sample weight, in grams (g).
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