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GB 23200.61-2016 English PDF

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GB 23200.61-2016: Food safety national standard -- Determination of benzidine residues in food by gas chromatography-mass spectrometry
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Basic data

Standard ID GB 23200.61-2016 (GB23200.61-2016)
Description (Translated English) Food safety national standard -- Determination of benzidine residues in food by gas chromatography-mass spectrometry
Sector / Industry National Standard
Classification of Chinese Standard G25
Word Count Estimation 13,146
Date of Issue 2016-12-18
Date of Implementation 2017-06-18
Older Standard (superseded by this standard) SN/T 2456-2010
Regulation (derived from) State Health Commission, Ministry of Agriculture, Food and Drug Administration Notice No. 16 of 2016
Issuing agency(ies) National Health and Family Planning Commission of the People's Republic of China, State Food and Drug Administration

GB 23200.61-2016: Food safety national standard -- Determination of benzidine residues in food by gas chromatography-mass spectrometry


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Food safety national standard - Determination of benzidine residues in food by gas chromatography - mass spectrometry National Standards of People's Republic of China GB Replace SN/T 2456-2010 National standards for food safety Determination of benzidine residues in food Gas chromatography - mass spectrometry National food safety standards- Determination of propham residue in foods Gas chromatography-mass spectrometry 2016-12-18 Release.2017-06-18 Implementation National Health and Family Planning Commission of the People 's Republic of China Issued by the Ministry of Agriculture of the People 's Republic of China State Administration of Food and Drug Administration

Foreword

This standard replaces SN/T 2456-2010 "Determination of benzidine residues in food for import and export by gas chromatography-mass spectrometry". Compared with SN/T 2456-2010, the main changes are as follows. - Standard text format is modified to national standard text format for food safety; - the name of the "import and export food" to "food"; - increase the "other food reference implementation" in the standard range. This standard replaced the previous version of the standard release. -SN/T 2456-2010. National standards for food safety Determination of benzidine residues in food by gas chromatography - mass spectrometry

1 Scope

This standard specifies the method for the determination of alanil residues in food by gas chromatography-mass spectrometry. This standard applies to soybeans, mustard, citrus, peanuts, tea, ginger, mushrooms, eels, pork, chicken liver, rice, soybeans and so on Determination and confirmation of the residues of amphetamines in export foodstuffs, and other foods may be referred to.

2 normative reference documents

The terms of the following documents are hereby incorporated by reference into this standard. Whichever is the date of the reference file, which is followed by all The amendments (not including corrigenda) or revisions do not apply to this standard, however, encourage the parties to reach an agreement under this standard Whether you can use the latest version of these files. For undated references, the latest edition applies to this standard. GB 2763 Maximum residue limit for pesticide in food GB/T 6682 Analytical laboratory water specifications and test methods

3 principle

The residual aniline in the sample was extracted with a mixed solvent of ethyl acetate-n-hexane (1 1, V/V). The plant-derived food was treated with graphite carbon black solid phase Extraction of pellets, animal food or high fat content of food by gel permeation chromatography (GPC) purification, gas chromatography - mass spectrometry Quantity and external standard method.

4 reagents and materials

Unless otherwise stated, the reagents used are pesticide residues and water is the primary water specified in GB/T 6682. 4.1 Reagents 4.1.1 n-hexane (C6H14). 4.1.2 Ethyl acetate (C4H8O2). 4.1.3 Sodium chloride (NaCl), analytical grade. 4.1.4 Cyclohexane (C6H12). 4.2 solution preparation 4.2.1 Ethyl acetate - n-hexane (1 1, V/V) Mixed solvent. 100 mL of ethyl acetate and 100 mL of n-hexane were added and mixed. 4.2.2 saturated sodium chloride solution. sodium chloride water dubbed saturated solution. 4.2.3 Ethyl acetate-cyclohexane (1 1, V/V) Mixed solvent. 100 mL of ethyl acetate and 100 mL of cyclohexane were added and mixed. 4.3 standards 4.3.1 Pesticide standards. Proparam (CAS. 122-42-9, C10H13NO2) Standard purity ≥ 95%. 4.4 standard solution preparation 4.4.1 Standard stock solution (100 mg/L). Accurately weigh the standard 10.0 mg, dissolve with n-hexane and set the volume to 100 mL. The stock solution was placed in a 4 ° C refrigerator. 4.4.2 standard working fluid. according to the need to take appropriate standard stock solution, n-hexane diluted to the appropriate concentration of standard working fluid. Standard work Liquid to now with the use. 4.5 Materials 4.5.1 ENVI-Carb Graphite Carbon Black Solid Phase Extraction Column. 250 mg, 3 mL, or equivalent. 4.5.2 GPC column packing. 20g.200 ~ 400 mesh Bio-beads S-X3.

5 instruments and equipment

5.1 Gas Chromatograph. with quality detector. 5.2 Gel Permeation Chromatography. 5.3 Crusher. 5.4 Tissue crusher. 5.5 Scroll Mixer. 5.6 Analytical balance. 0.01 g and 0.0001 g. 5.7 screw cap polytetrafluoroethylene plastic centrifuge tube, 50mL. 5.8 Centrifuge. 2500r/min. 5.9 Pipettes. 1 mL to 5 mL, 100 μL to 1 000 μL.

6 Preparation and storage of samples

6.1 Preparation of the sample 6.1.1 Fruits and vegetables Take representative fruits such as citrus, mustard, soybeans, ginger, peanuts and other fruits and vegetables at least 500 g, (not water washable) Tissue crusher The sample is processed into a slurry, mixed, divided into two as a sample, divided into a clean sample bag, sealed and identified. 6.1.2 rice, soybeans, tea, edible fungi (dry) class Take at least 300 g of representative samples such as tea leaves, edible fungi (dry) and the like, crushed with a pulverizer and passed through a 2.0 mm round hole sieve, Are divided into two as a sample, into the clean Sheng Sheng bag, sealed and identified. 6.1.3 Meat and viscera Take at least 500 g of representative samples of pork, chicken, fish and other meat and meat products. After chopping, the sample is processed into a Paste, mix, are divided into two as a sample, into the clean Sheng Sheng bag, sealed and identified. Note. The above sample sampling site according to GB 2763 Appendix A implementation. 6.2 Sample storage The sample should be stored at -18 ° C; during sample preparation, the sample should be protected from contamination or changes in the residue content.

7 Analysis steps

7.1 Extraction 7.1.1 Fruits and vegetables Weigh 5g (accurate to 0.01g) homogeneous sample, placed in 50 mL stoppered plastic centrifuge tube, add sodium chloride 3 g, then add 10 mL B Acid ethyl ester-n-hexane mixed solvent vortex shaker for 30 s, the ultrasonic extraction 20 min, 2500 r/min centrifugation 3 min, take the upper Organic phase in another test tube, and then were added 5 mL ethyl acetate - n-hexane mixed solvent repeated extraction 2 times, combined extract, 40 ℃ Nitrogen was purged to about 1 mL and purified according to 7.2.1. 7.1.2 tea, rice, edible fungi (dry) class Weigh 2.5 g (accurate to 0.01 g) homogeneous sample placed in a 50 mL stoppered plastic centrifuge tube, add 2.5 mL of saturated sodium chloride solution, Add 5 mL of ethyl acetate-n-hexane mixed solvent, vortex oscillator for 30 s, ultrasonic extraction 20 min, centrifuge at 2500r/min 3 min, Take the upper organic phase in another tube. And then added ethyl acetate - n-hexane mixed solvent were extracted twice, combined extract, 40 ℃ Nitrogen was purged to about 1 mL and purified according to 7.2.2. 7.1.3 Soybean, meat and viscera For samples of chicken liver, pork, fish and other samples weighed 5g (accurate to 0.01g) uniform sample placed in 50 mL stopper plastic centrifuge tube, plus Sodium chloride 3 g, then add 10 mL mixed solvent vortex oscillator for 30 s, after ultrasonic extraction 20 min, 2500 r/min centrifugal 3 min, Take the upper organic phase in another tube, and then add 5 mL of ethyl acetate - n-hexane mixed solvent repeated extraction 2 times, combined extract, 40 ℃ nitrogen blowing concentrated to 10.0 mL, according to 7.2.3 purification steps. 7.2 Purification 7.2.1 Samples extracted in accordance with 7.1.1 The ENVI-Carb column was loaded on a solid phase extraction vacuum suction apparatus and the mixture was washed with 3 mL of ethyl acetate-n-hexane mixed solvent Column, the flow rate is about 1 mL/min. The extract was passed through an ENVI-Carb column and eluted with 6 mL of a mixture of ethyl acetate-n-hexane, The whole eluate was collected and allowed to stand at 40 ° C to near dryness and 2.0 mL with ethyl acetate for GC-MS analysis. 7.2.2 Samples extracted in accordance with 7.1.2 The ENVI-Carb column was loaded on a solid phase extraction vacuum suction apparatus and the mixture was washed with 3 mL of ethyl acetate-n-hexane mixed solvent Column, the flow rate is about 1 mL/min. The extract was passed through an ENVI-Carb column and eluted with 6 mL of a mixture of ethyl acetate-n-hexane, The whole eluate was collected and the nitrogen was blown to near dry at 40 ° C and 1.0 mL in volume with ethyl acetate for GC-MS analysis. 7.2.3 Samples extracted in accordance with 7.1.3 5 mL of the extract was passed through a GPC column and eluted with a mixed solvent of ethyl acetate-cyclohexane (1 1, V/V) to maintain a flow rate of 4.7 ml/min, The 30 ~ 43 mL eluate was collected, concentrated to near dryness, and quantified to 1.0 mL with ethyl acetate for GC-MS analysis. 7.3 Determination 7.3.1 Gas Chromatography - Mass Spectrometry Reference Conditions A) Column. DB-5MS quartz capillary column, 30 m x 0.25 mm (id) x 0.25 μm, or equivalent; B) Column temperature. initial temperature 80 ° C, rise to 130 ° C at 10 ° C/min, rise to 136 ° C at 0.5 ° C/min, At 40 ° C/min up to 280 ° C for 2 min; C) Inlet temperature. 260 ° C; D) ion source. 230 ° C; E) Select the monitoring ion (m/z). Quantitative ion 120; Qualitative ions 93,137,179 F) Carrier gas. helium, purity greater than or equal 99.999%, flow rate 1 mL/min; G) Injection volume. 1 μL; H) Injection method. Splitless injection I) Quadrupole temperature. 250 ° C; J) Connection temperature. 280 ° C 7.3.2 Determination and confirmation of chromatography According to the content of the sample in the sample, select the appropriate response value of the standard working solution for chromatographic analysis of the standard working fluid and sample And so on. The response value of the benzamide in the standard working fluid and the sample to be measured shall be within the linear response range of the instrument. If the sample solution is in the selective ion chromatogram of the standard working solution, a chromatographic peak appears at the same retention time, and after subtracting the background In the sample mass spectrum, m/z120,93,137,179 appears, the abundance of the selected ions was higher than the abundance ratio of the corresponding ions Within the permissible range (see Table 1 for the permissible range), it is possible to determine the presence of aniline residues in the sample. Under the above conditions of the instrument, aniline Reference retention time is 14.2 min, ion abundance ratio m/z93. 120. 137. 179 = 100. 34. 42. 54. Table 1 Maximum allowable error for relative ion abundance using gas chromatography-mass spectrometry 7.4 blank experiment In addition to the sample, according to the above determination steps.

8 results are calculated and expressed

Use the chromatographic data processor or calculate the content of alanine in the sample by the following formula (1). MAs FVCsA   (1) Where. X - Pesticide residue in milligrams per kilogram (mg/kg); The peak area of pesticide in A sample solution; Cs - the concentration of pesticide in the standard working fluid, in milligrams per liter (mg/L); V - the final volume of the sample, in milliliters (mL); F - dilution or concentration multiple SA - peak area of pesticide in standard working fluid; M - the amount of sample, in grams (g). Note. The result of the calculation shall be deducted from the blank value. The result of the measurement shall be expressed as the arithmetic mean of the parallel measurement, and two valid digits shall be retained.

9 precision

9.1 The ratio of the absolute difference between the two independent determinations obtained under reproducibility and its arithmetic mean (percentage) shall be in accordance with Appendix E requirements. 9.2 The ratio of the absolute difference between the two independent determinations obtained under reproducibility and its arithmetic mean (percentage) shall be in accordance with Appendix F requirements. 10% limit and recovery rate Relative abundance (base) 50% 20% to 50% 10% to 20% ≤10% Allowable relative deviation ± 10% ± 15% ± 20% ± 50% 10.1 Quantitation limits The quantification limit of this method is 0.005 mg/kg. 10.2 Recovery rate When the levels were 0.005 mg/kg, 0.01 mg/kg, 0.02 mg/kg, 0.05 mg/kg, the aniline pesticides were present in different substrates The recovery rate is given in Appendix D.

Appendix A

(Informative) Aniline pesticide CAS number, retention time and monitoring ion abundance ratio Pesticide Chinese name Pesticide English name CAS number Molecular formula Molecular retention time Min Monitor the ion abundance ratio 1 aniline spirit propham 122-42-9 C10H13NO2 179.22 14.2 93 * (100), 120 (34), 137 (42), 179 (54) Note." "Marked ions are quantitative ions

Appendix B

(Informative) Optimum chromatographic extraction of aniline

Appendix C

(Informative) Phenylamine standard quality spectrum

Appendix D

(Informative) Addition Recovery Rate of Aniline Pesticides in Different Substrates Sample concentration, mg/kg recovery range,% sample concentration, mg/kg recovery range,% Soybeans 0.005 76.0% to 106.0% Mustard 0.005 68.0% to 96.0% 0.01 77.0% to 104.0% 0.01 72.0% to 100.0% 0.02 74.0% ~ 112.5% 0.02 76.0% ~ 109.0% 0.05 71.0% to 95.8% 0.05 86.8% to 101.2% Citrus 0.005 86.0% to 108.0% peanut 0.005 70.0% to 90.0% 0.01 90.0% to 110.0% 0.01 68.0% to 99.0% 0.02 77.5% to 110.5% 0.02 77.5% to 112.5% 0.05 70.6% to 87.0% 0.05 89.4% to 98.8% tea 0.005 74.0% to 110.0% 0.005 70.0% to 100.0% 0.01 73.0% to 105.0% 0.01 73.0% to 106.0% 0.02 80.0% to 109.0% 0.02 77.0% to 101.0% 0.05 70.0% to 107.2% 0.05 71.6% to 99.0% Mushrooms 0.005 64.0% to 100.0% eel 0.005 88.0% to 96.0% 0.01 72.0% to 99.0% 0.01 74.0% to 105.0% 0.02 74.0% to 108.5% 0.02 78.5% to 90.5% 0.05 66.0% to 91.4% 0.05 73.6% to 88.4% pork 0.005 84.0% ~ 96.0% Chicken liver 0.005 78.0% to 96.0% 0.01 75.0% to 86.0% 0.01 72.0% to 86.0% 0.02 75.5% ~ 87.5% 0.02 75.5% ~ 85.5% 0.05 72.4% to 80.6% 0.05 71.2% to 75.8% Rice 0.005 84.0% to 104.0% Soybeans 0.005 74.0% to 98.0% 0.01 75.0% ~ 94.0% 0.01 79.0% ~ 102.0% 0.02 83.0% to 100.5% 0.02 78.0% to 101.0% 0.05 71.6% to 94.0% 0.05 73.4% to 95.8%

Appendix E

(Normative appendix) Laboratory repeatability requirements Table E.1 Laboratory repeatability requirements Measured component content Mg/kg Precision 0.001 36 > 0.01 > 1 14

Appendix F

(Normative appendix) Inter-laboratory reproducibility requirements Table F.1 Inter-laboratory reproducibility requirements Measured component content Mg/kg Precision 0.001 54 > 0.01 > 1 19

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