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GB 5009.265-2021 English PDF

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GB 5009.265-2021: National food safety standard - Determination of polycyclic aromatic hydrocarbons in foods
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GB 5009.265: Evolution and historical versions

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GB 5009.265-2021English439 Add to Cart 4 days [Need to translate] National food safety standard - Determination of polycyclic aromatic hydrocarbons in foods Valid GB 5009.265-2021
GB 5009.265-2016English85 Add to Cart 0--9 seconds. Auto-delivery National food safety standard -- Determination of polycyclic aromatic hydrocarbons in foods Obsolete GB 5009.265-2016

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Basic data

Standard ID GB 5009.265-2021 (GB5009.265-2021)
Description (Translated English) National food safety standard - Determination of polycyclic aromatic hydrocarbons in foods
Sector / Industry National Standard
Classification of Chinese Standard X09
Word Count Estimation 22,232
Issuing agency(ies) National Health Commission of the People's Republic of China, State Administration for Market Regulation

GB 5009.265-2021: National food safety standard - Determination of polycyclic aromatic hydrocarbons in foods


---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
(National Food Safety Standard Determination of Polycyclic Aromatic Hydrocarbons in Food) National Standards of People's Republic of China National Food Safety Standard Determination of polycyclic aromatic hydrocarbons in food Released on 2021-09-07 Implementation on 2022-03-07 National Health Commission of the People's Republic of China Issued by the State Administration for Market Regulation National Food Safety Standard Determination of polycyclic aromatic hydrocarbons in food

1 Scope

The first method of this standard specifies the gas chromatography-mass spectrometry method for the determination of 16 kinds of polycyclic aromatic hydrocarbons in food; the second method specifies 15 kinds of polycyclic aromatic hydrocarbons in food Method for determination of polycyclic aromatic hydrocarbons by liquid chromatography. The first method of this standard applies to 16 types of polycyclic foods and their products, meat and their products, aquatic products and their products, animal and vegetable fats and their products. Aromatic hydrocarbons (benzo[c]fluorene, benzo[a]anthracene, cyclopenta[c,d]pyrene, 5-methyl, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo [j]Fluoranthene, Benzo[a]pyrene, indeno[1,2,3-c,d]pyrene, dibenzo[a,h]anthracene, benzo[g,h,i]perylene, dibenzo[a,l ] Pyrene, dibenzo[a,e] pyrene, di Determination of benzo[a,i]pyrene, dibenzo[a,h]pyrene); suitable for four kinds of polycyclic aromatic hydrocarbons (benzo [a] Determination of anthracene, benzo[b]fluoranthene, benzo[a]pyrene). The second method of this standard applies to 15 kinds of polycyclic aromatics in grain and its products, meat and its products, aquatic products and its products, vegetables, and animal and vegetable oils. Hydrocarbons (naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno [1,2,3-c,d] Determination of pyrene, dibenzo[a,h]anthracene and benzo[g,h,i]perylene). Method 1 Gas Chromatography-Mass Spectrometry

2 Principle

The polycyclic aromatic hydrocarbons in the sample are extracted by solvent, saponified with potassium hydroxide ethanol solution, purified by solid phase extraction column, and concentrated by gas chromatography-mass spectrometry. Measured with instrument and quantified by isotope internal standard method.

3 Reagents and materials

Unless otherwise specified, the reagents used in this method are all analytically pure, and the water is the first grade water specified in GB/T 6682. 3.1 Reagents 3.1.1 Ethyl acetate (C4H8O2). chromatographically pure. 3.1.2 Dichloromethane (CH2Cl2). chromatographically pure. 3.1.3 Acetone (C3H6O). chromatographically pure. 3.1.4 Cyclohexane (C6H12). chromatographically pure. 3.1.5 n-hexane (C6H14). chromatographically pure. 3.1.6 Isooctane (C8H18). chromatographically pure. 3.1.7 Absolute ethanol (C2H6O). chromatographically pure. 3.1.8 Ammonia (NH3·H2O). The mass fraction is about 25%. 3.1.9 Anhydrous ether (C4H10O). 3.1.10 Petroleum ether (C5H12O2). the boiling range is 30℃~60℃. 3.1.11 Potassium hydroxide (KOH). 3.1.12 Anhydrous sodium sulfate (Na2SO4). Bake at 400℃ for 2h before use, then put it in the desiccator for later use. 3.2 Reagent preparation 3.2.1 Potassium hydroxide ethanol solution (0.3mol/L). Weigh 1.68g potassium hydroxide, dissolve it with 100mL absolute ethanol ultrasonically, and use it immediately Now equipped. 3.2.2 Potassium hydroxide ethanol solution (1.5mol/L). Weigh 8.40g potassium hydroxide, dissolve it with 100mL absolute ethanol ultrasonically, and use it immediately Now equipped. 3.2.3 Cyclohexane-ethyl acetate solution (11). Mix cyclohexane and ethyl acetate according to 1.1 (volume ratio) evenly. 3.2.4 Dichloromethane-ethyl acetate solution (11). Mix dichloromethane and ethyl acetate according to 1.1 (volume ratio) evenly. 3.2.5 Acetone-Isooctane Solution (1 1). Mix acetone and isooctane according to 1.1 (volume ratio) evenly. 3.3 Standard solution 3.3.1 PAH standard solution Polycyclic aromatic hydrocarbon standard solution (containing benzo[c]fluorene, benzo[a]anthracene, cyclopenta[c,d]pyrene, 5-methyl, benzo[b]fluoranthene, benzo [k]fluoranthene, benzo[j]fluoranthene, benzo[a]pyrene, indeno[1,2,3-c,d]pyrene, dibenzo[a,h]anthracene, benzo[g,h,i] perylene, dibenzo[a,l] Pyrene, dibenzo[a,e]pyrene, dibenzo[a,i]pyrene, dibenzo[a,h]pyrene, 10.0μg/mL or corresponding concentration). certified by the country and awarded the standard The standard solution of the quasi-substance certificate should be stored at -18℃ and protected from light. See Appendix A for the CAS numbers of PAHs. 3.3.2 Isotope polycyclic aromatic hydrocarbon internal standard solution Isotope polycyclic aromatic hydrocarbon internal standard solution (containing at least D12-benzo[a]anthracene, D12-, D12-benzo[b]fluoranthene, D12-benzo[a]pyrene, D12-indene And [1,2,3-c,d]pyrene, D14-dibenzo[a,h]anthracene, D12-benzo[g,h,i]perylene, 100.0μg/mL or corresponding concentration), -18 Avoid under ℃ Light save. See Appendix A for the internal standard CAS number of isotope polycyclic aromatic hydrocarbons. Warning---PAHs are known carcinogenic, teratogenic, and mutagenic substances, and their carcinogenicity increases with the number of benzene rings. Enter Particular attention should be paid to safety protection when performing standard solution preparation and other operations. The operation should be performed in a fume hood to minimize exposure. 3.4 Preparation of standard liquid 3.4.1 PAH standard solution (500.0ng/mL). draw 0.50mL of PAH standard solution (10.0μg/mL) in 10mL In the volumetric flask, dilute to the mark with the acetone-iso-octane solution (1 1), mix well, transfer the solution to a brown glass storage bottle, and protect from light at 4°C Sealed and stored, the shelf life is 3 months. 3.4.2 PAH standard use solution (100.0ng/mL). Pipette 2.0mL of PAH standard use solution (500.0ng/mL) in 10mL In the volumetric flask, dilute to the mark with the acetone-iso-octane solution (1 1), mix well, transfer the solution to a brown glass storage bottle, and protect from light at 4°C Sealed and stored, the shelf life is 3 months. 3.4.3 Isotope polycyclic aromatic hydrocarbons internal standard solution (200.0ng/mL). draw the isotope polycyclic aromatic hydrocarbons internal standard solution (100.0μg/mL) 0.10mL in a 50mL volumetric flask, dilute to the mark with acetone-iso-octane solution (1 1), mix well, transfer the solution to brown glass for storage In the bottle, store it in a sealed container at 4℃ in the dark, and the shelf life is 1 year. 3.4.4 PAH standard series working fluid. Take 8 cone-bottomed sample bottles, and draw the PAH standard fluid (100.0ng/mL) 20.0μL, 40.0μL, 50.0μL, 80.0μL and PAH standard use fluid (500.0ng/mL) 20.0μL, 40.0μL, 50.0μL, 100.0μL, then each add isotope polycyclic aromatic hydrocarbons internal standard solution (200.0ng/mL) 100.0μL, supplemented with acetone-iso-octane solution (1 1) The volume is up to.200μL, and the concentration of the standard series of PAHs is 10.0ng/mL, 20.0ng/mL, 25.0ng/mL, 40.0ng/mL, 50.0ng/mL, 100.0ng/mL, 125.0ng/mL, 250.0ng/mL, the internal standard is 100.0ng/mL, ready for immediate use.

4 Apparatus and equipment

4.1 Gas chromatography-quadrupole mass spectrometer (GC-MS). equipped with electron bombardment ion source (EI source). 4.2 Electronic balance. Sensitivity is 0.01g, 0.001g. 4.3 High-speed crusher. 4.4 Homogenizer. 4.5 Constant temperature water bath device. 4.6 Ultrasonic cleaner. 4.7 Nitrogen blowing instrument. 4.8 High-speed centrifuge. speed ≥10000r/min. 4.9 Vortex oscillator. 4.10 Precision pipette. 4.11 Solid phase extraction cartridge (styrene divinylbenzene polymer) or equivalent purification column. 6mL, 300mg.

5 Analysis steps

5.1 Sample preparation 5.1.1 Sample preparation 5.1.1.1 Grain and its products Take a sample of.200g~500g, use a high-speed grinder to fully crush it, mix it evenly, and separate it into about 100g, and pack it in a clean sample bag Or in the bottle, sealed and stored at -18℃ for testing. 5.1.1.2 Meat and its products, aquatic products and its products, vegetables Take a sample of.200g~500g, chop the edible part first, then fully mash it with a homogenizer, take about 100g and put it in a clean sample bag or In the bottle, it should be frozen and stored at -18℃ after sealing the label for testing. 5.1.1.3 Animal and vegetable oils and products Take 100g~200g of animal and vegetable oils and products in bulk, and put them in clean sample bottles, seal them and store them in a cool place for testing. The final packaging of animal and vegetable oils is labelled and stored in a cool place for testing. 5.1.1.4 Infant formula milk powder, supplementary food for infants and young children Take.200g~500g of the sample to be crushed, use a high-speed crusher to fully crush it, mix evenly, and separate out about 100g, and install it in a clean In the sample bag or bottle, store it in a refrigerator at 4℃ after sealing the label for testing. The infant formula milk powder and infant supplementary food that do not need to be crushed are labeled and stored in a cool place for testing. 5.1.2 Sample extraction 5.1.2.1 Grain and its products Weigh 5g of the sample (accurate to 0.001g), place it in a 50mL centrifuge tube with a stopper, and add 100μL of isotope polycyclic aromatic hydrocarbon internal standard for use Solution (200.0ng/mL) shake and mix and let stand for 30min, add 5g anhydrous sodium sulfate, 20mL cyclohexane-ethyl acetate mixed solution (1 1) Vortex extraction for 1min, ultrasonic extraction for 15min, centrifuge at 10000r/min for 3min, and draw the supernatant to 15mL pre-weighed centrifugation In a tube (accurate to 0.01g), dry the solvent with nitrogen in a water bath at 45°C to obtain the extract, weigh it, calculate the mass of the extract (accurate to 0.01g), and extract Material to be saponified. If the mass of the extract is greater than 1g, weigh out 1g of the extract (accurate to 0.01g) and place it in another 15mL centrifuge tube with a stopper for saponification. 5.1.2.2 Meat and its products, aquatic products and its products Weigh 5g of the sample (accurate to 0.001g), place it in a 50mL centrifuge tube with a stopper, and add 100μL of isotope polycyclic aromatic hydrocarbon internal standard for use Liquid (200.0ng/mL) shake and mix and let stand for 30min, add 20g anhydrous sodium sulfate, stir and disperse with a metal spoon, add 20mL ring Hexane-ethyl acetate mixed solution (11), vortex extraction for 1 min, further ultrasonic extraction for 15 min, centrifuge at 10000r/min for 3 min, suction Take the supernatant into a 15mL pre-weighed centrifuge tube, dry the solvent with nitrogen under a water bath at 45°C to obtain the extract, weigh it, and calculate the quality of the extract (Accurate to 0.01g), the extract is to be saponified. If the mass of the extract is greater than 1g, weigh out 1g of the extract (accurate to 0.01g) and place it in another 15mL centrifuge tube with a stopper for saponification. 5.1.2.3 Animal and vegetable oils and products Weigh 0.5g~1g of the sample (accurate to 0.001g), place it in a 15mL centrifuge tube with a stopper, and add 100μL of isotope polycyclic aromatic hydrocarbons Standard use solution (200.0ng/mL), to be saponified. 5.1.2.4 Infant formula milk powder Weigh 2g of the sample (accurate to 0.001g), place it in a 50mL centrifuge tube with a stopper, and add 100μL of isotope polycyclic aromatic hydrocarbon internal standard for use Solution (200.0ng/mL), add 10mL water, vortex to dissolve, add 2mL ammonia water to mix well, place in 65℃±5℃ water bath for 10min, Take out and cool to room temperature. Add 10mL of absolute ethanol, mix slowly, add 8mL of anhydrous ether, vortex for 5min, then add 8mL Petroleum ether, vortex for 5min, centrifuge at 10000r/min for 5min, aspirate the upper organic phase into a 15mL pre-weighed centrifuge tube, Dry the solvent with nitrogen under a water bath at 45℃ to obtain the extract, weigh it, and calculate the mass of the extract (accurate to 0.01g), and the extract is to be saponified. 5.1.2.5 Supplementary foods for infants and young children Weigh 2g (accurate to 0.001g) sample, place it in a 50mL centrifuge tube with stopper, add 100μL isotope polycyclic aromatic hydrocarbon internal standard for use Solution (200.0ng/mL), add 10mL water, 10mL absolute ethanol, vortex to dissolve, add 8mL anhydrous ether, vortex for 5min, then Add 8mL petroleum ether, vortex for 5min, centrifuge at 10000r/min for 5min, aspirate the upper organic phase to 15mL pre-weighed In the heart tube, dry the solvent with nitrogen in a water bath at 45℃ to obtain the extract, weigh it, and calculate the mass of the extract (accurate to 0.01g). The extract is to be saponified. 5.1.3 Saponification 5.1.3.1 If the mass of the extract is less than or equal to 0.2g (accurate to 0.01g), add 5mL of 0.3mol/L potassium hydroxide ethanol solution, vortex to mix Homogenize, place at room temperature for 5 min, add 4 mL water, 5 mL n-hexane, vortex for 2 min, centrifuge at 10000r/min for 2 min, and extract the upper layer with n-hexane. Take the liquid to be purified. 5.1.3.2 If the mass of the extract is between 0.2g~1g (accurate to 0.01g), add 5mL 1.5mol/L potassium hydroxide ethanol solution, and cover Vortex to mix, place in a 70℃±2℃ water bath for saponification for 3min. After the saponification is complete, take it out and cool it to room temperature with tap water, add 4 mL of water, 5mL n-hexane, vortex extraction for 2min, centrifugation at 10000r/min for 2min, the upper n-hexane extract is to be purified. 5.1.3.3 Add 5 mL of 1.5 mol/L potassium hydroxide ethanol solution to the animal and vegetable oil sample (the solid oil sample is first placed in a 50 ℃ water bath to melt Add after chemical conversion), cover and vortex to mix, place in 70℃±2℃ water bath for saponification for 3min. After the saponification is completed, take it out and cool it to the room with tap water Warm, add 4mL water, 5mL n-hexane, vortex extraction for 2min, centrifuge at 10000r/min for 2min, the upper n-hexane extract is to be purified. Note. Benzo [c] fluorene is unstable to alkali. When measuring benzo [c] fluorene, the saponification time depends on the quality of the oil, and 1g extract is saponified for 3 minutes. 5.1.4 Purification Add about 1 g of anhydrous sodium sulfate to the solid phase extraction cartridge, and wash the activated column with 3 mL of dichloromethane and 3 mL of n-hexane in sequence. After the completion of the chemistry, all the n-hexane extract is transferred to the solid phase extraction column, and the flow rate is controlled at about 1.0 mL/min. After passing through the packing layer, first rinse with 5 mL n-hexane to remove impurities, and then eluted with 5 mL dichloromethane-ethyl acetate solution (1 1), and collect the eluent In a 10mL conical bottom glass test tube. The eluent was blown dry with nitrogen in a water bath at 40°C, and then vortexed with 0.1 mL of acetone-iso-octane solution (1 1) Spin to dissolve the residue and transfer it to the cone-bottom sample bottle for GC-MS determination. 5.1.5 Blank test Except that no sample is added, the analysis procedure is exactly the same as that of the sample. 5.1.6 Parallel test According to the above steps, the same sample is tested in parallel. 5.2 Instrument reference conditions 5.2.1 Reference conditions for gas chromatography. 5.2.1.1 Chromatographic column. DB-EUPAH capillary column. column length 20m, inner diameter 0.18mm, film thickness 0.14μm or equivalent chromatographic column. 5.2.1.2 The temperature of the injection port. 280℃. 5.2.1.3 Carrier gas. helium, purity ≥99.999%. 5.2.1.4 Injection volume. 1μL~2μL, splitless injection. 5.2.1.5 Solvent delay. 16.5min. 5.2.1.6 Column temperature program. initial temperature 80℃, keep for 2min, increase to 250℃ at 10℃/min, keep for 2min, increase to 8℃/min Keep it at 315℃ for 5min, and finally raise it to 320℃ at 20℃/min and keep it for 5min. 5.2.1.7 Flow program. 0.7mL/min is maintained for 32min, and then 5mL/min is increased from 0.7mL/min to 1.5mL/min to the end. 5.2.2 Reference conditions for mass spectrometry. 5.2.2.1 Ionization method. electron impact source (EI). 5.2.2.2 The ionization energy is. 70eV. 5.2.2.3 Ion source temperature. 230℃. 5.2.2.4 Transmission line temperature. 280℃. 5.2.2.5 Quadrupole temperature. 150℃. 5.2.2.6 Measurement method. select ion monitoring method. 5.2.2.7 Monitoring ion. see Appendix B. 5.3 Qualitative analysis The retention time of the chromatographic peak of the target compound in the sample is compared with the retention time of the chromatographic peak of the corresponding standard target compound, and the range of change is Should be within ±0.5%. The relative abundance of the target compound monitoring ion in the sample is consistent with the relative abundance of the standard solution at the same concentration, and the allowable deviation does not exceed According to the provisions of Table 1, it can be judged that there is a corresponding measured object in the sample.

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