Standard Briefing:Stadard ID: GB 23200.77-2016
Stadard Title: Food safety national standard -- Detection Method of Benzomidazole Residue in Food
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Food safety national standard -- Detection Method of Benzomidazole Residue in Food
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GB 23200.77-2016
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Basic Data: | Standard ID | GB 23200.77-2016 (GB23200.77-2016) | | Description (Translated English) | Food safety national standard -- Detection Method of Benzomidazole Residue in Food | | Sector / Industry | National Standard | | Word Count Estimation | 12,132 | | Date of Issue | 2016-12-18 | | Date of Implementation | 2017-06-18 | | Older Standard (superseded by this standard) | SN/T 2431-2010 | | Regulation (derived from) | State Health Commission, Ministry of Agriculture, Food and Drug Administration Notice No. 16 of 2016 | Contents, Scope, and Excerpt:GB 23200.77-2016
Food safety national standard - Detection Method of Benzomidazole Residue in Food
National Standards of People's Republic of China
GB
Replace SN/T 2431-2010
National standards for food safety
Detection Method of Benzomidazole Residue in Food
National food safety standards-
Determination of halfenprox residue in foods
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
Preface
This standard replaces SN/T 2431-2010 "Determination of benzalkalate residues in food for import and export".
Compared with SN/T 2431-2010, the main changes are as follows.
- Standard text format is modified to national standard text format for food safety;
- the name and scope of the "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 2431-2010.
National standards for food safety
Detection Method of Benzomidazole Residue in Food
Scope
This standard specifies the determination of benzimidazole residues in food by gas chromatography and gas chromatography-mass spectrometry.
This standard applies to asparagus, potatoes, onions, pears, peaches, corn, buckwheat, tea, vinegar, honey, walnut, rabbit, chicken
Liver, shrimp, chicken in the detection of benzalkalcone residue, other food can refer to the implementation.
2 normative reference documents
The following documents are indispensable for the application of this document. For dated references, only the dated edition applies to this
file. For undated references, the latest edition (including all modifications) applies to this document.
GB 2763 National Standard for Food Safety - Maximum Residue Limit of Pesticides in Foodstuffs
GB/T 6682 Analytical laboratory water specifications and test methods
3 principle
Vinegar and honey dissolved and diluted with water, the solid phase extraction column purification; onion and tea with ethyl acetate - cyclohexane mixed solvent extraction, solid
Phase extraction and purification; animal food and high oil content of food with acetonitrile extraction, other food with ethyl acetate - cyclohexane mixed solvent extraction,
The extract was chromatographed on a gel chromatograph (GPC) with an electron capture detector, quantified by external standard method, positive sample gas
Determination by phase chromatography - mass spectrometry.
4 reagents and materials
Unless otherwise specified, all reagents are of analytical grade and water is in accordance with the primary water specified in GB/T 6682.
4.1 Reagents
4.1.1 Ethyl acetate (C4H8O2, CAS No. 141-78-6). Chromatographic purity.
4.1.2 Cyclohexane (C6H12, CAS No. 110-82-7). Chromatographic purity.
4.1.3 Acetonitrile (C2H3N, CAS No. 75-05-8). Chromatographic Purification.
4.1.4 n-hexane (C6H14, CAS No. 110-54-3). chromatographic purity
4.1.5 Acetone (C3H6O, CAS No. 67-64-1). Chromatographic Purification
4.1.6 anhydrous sodium sulfate (Na2SO4, CAS No. 15124-09-1). 650 ℃ burning 4 h, stored in a sealed container for use.
4.1.7 Sodium chloride (NaCl, CAS No. 7647-14-5).
4.2 solution preparation
4.2.1 Ethyl acetate-cyclohexane mixed solvent (1 1, V/V). 500 mL of ethyl acetate and 500 mL of cyclohexane were added and mixed.
4.2.2 N-Hexane - Acetone Mixed Solvent (9 1, V/V). Take 90 mL of n-hexane and 10 mL of acetone.
4.2.3 Cyclohexane-ethyl acetate mixed solvent (6 1, V/V). Take 10 mL of ethyl acetate and 60 mL of cyclohexane.
4.3 standards
4.3.1 Benzylidene ether Reference material. (Halfenprox; CAS. 111872-58-3; C24H23BrF2O3; Molecular weight. 477.34) Purity
≥99%.
4.4 standard solution preparation
4.4.1 standard stock solution (100 mg/L). accurately weighed the appropriate amount of standard material, dissolved with ethyl acetate, prepared into a concentration of 100 mg/L
Of the standard stock solution, the solution in 0-4 ℃ refrigerator to save.
4.4.2 standard working solution. if necessary, then diluted with ethyl acetate to the appropriate concentration of standard working solution. Standard working fluid should be used with the current distribution.
4.5 Materials
4.5.1 Graphitized Carbon Black and PSA Mixing Column. 1 mL, 50 mg Graphitized Carbon Black 50 mgPSA, mixed with 3 mL of cyclohexane-ethyl acetate
Solvent activation.
4.5.2 HLB solid phase extraction column. 3 mL, 60 mg, or equivalent, with 2 mL of n-hexane-acetone mixed solution, 2 mL of acetone, 5 mL
Distilled water pre-leaching.
4.5.3 Organic phase filter. 0.45 m.
5 instruments and equipment
5.1 Gas chromatograph with electronic capture detector.
5.2 Gas Chromatography-Mass Spectrometer with Electronic Bombing Source (EI).
5.3 Analysis of the balance. the amount of 0.01 g.
5.4 Analysis of the balance. the sense of 0.0001 g.
5.5 Gel chromatograph with unit pump, fraction collector.
5.6 Centrifuge. 4000 r/min.
5.7 Rotary Evaporator.
5.8 anhydrous sodium sulfate column. 7.5 cm × 1.5 (id) cm glass column, built 5 cm high anhydrous sodium sulfate.
5.9 Scroll Mixer.
5.10 homogenizer.
5.11 nitrogen blowing instrument.
5.12 with a plug centrifuge tube. 50 mL, polytetrafluoroethylene.
6 Preparation and storage of samples
6.1 Preparation of the sample
6.1.1 Grain and tea
The sample was divided by a quarter to 500 g, and the whole was ground with an attritor. Mix, are divided into two as a sample, into the clean Sheng Sheng
Inside the bottle, sealed, marked with the mark.
6.1.2 Fruits and vegetables
Take 500 g of fruit or vegetable sample (not water wash), and then cut the sample into a slurry with a food masher.
Mix, are divided into two as a sample, into the clean Sheng Sheng bag, sealed, marked mark.
6.1.3 meat and meat products, nuts
A sample of about 1 kg was taken from all the samples taken and crushed evenly by the crusher and divided into two portions
Net container as a sample. Sealed and marked.
Note. The above sample sampling site according to GB 2763 Appendix A implementation.
6.2 Sample storage
Cereals, tea, nuts, honey and honey products samples stored at 0-4 ℃; other samples in -18 ℃ below frozen
save.
During the operation of the sample preparation, the sample should be protected from contamination or changes in the content of the residue.
7 Analysis steps
7.1 Extraction
7.1.1 Vegetables, fruits, grain
Vegetables, fruit, cereal weighed 10 g (accurate to 0.01 g) homogeneous sample, placed in 50 mL with a plug centrifuge tube, add 20 mL of acetic acid B
Ester-cyclohexane mixture solvent, 10000 r/min homogenate 60 s, centrifuged at 4000 r/min for 3 min, the upper organic phase was collected and the residue was washed with 20 mL
Ethyl acetate-cyclohexane, and the combined organic phases were combined and dehydrated over anhydrous sodium sulfate column. The mixture was collected in 150 mL of concentrated
Bottle, concentrated in a 40 ° C water bath to near dry, accurately add 10.0 mL of ethyl acetate - cyclohexane dissolved residue, and over 0.45 m filter
Membrane, to be gel chromatographic purification.
7.1.2 walnut, rabbit meat, chicken liver, shrimp, chicken
Weigh 10 g (accurate to 0.01 g) homogeneous sample, placed in 50 mL stoppered plastic centrifuge tube, add 20 mL of acetonitrile, 10 mL distillation
Water, 3 g sodium chloride, 10000 r/min homogenate 60 s, 4000 r/min centrifugation 3 min, the upper organic phase was collected and the residue was washed with 20 mL of acetonitrile
The organic phase was combined and dehydrated over anhydrous sodium sulfate column. The mixture was collected in a 150 mL concentration flask and rotated in a 40 ° C water bath
Concentrated to near dry, accurately add 10.0 mL of ethyl acetate - cyclohexane dissolved residue, and over 0.45 m filter, gel chromatography to be purified.
7.1.3 tea, onions
Weigh 2.5 g (accurate to 0.01 g) homogeneous sample, placed in 50 mL stoppered plastic centrifuge tube, add 15 mL of distilled water, put it aside for 1 h,
Into the 20 mL ethyl acetate-cyclohexane mixture solvent, 10000 r/min homogenate 60 s, 4000 r/min centrifugation 3 min, collecting the upper organic phase, residual
The residue was extracted once again with 20 mL of ethyl acetate-cyclohexane mixed solvent. The combined organic phases were combined and dehydrated over anhydrous sodium sulfate.
In 150 mL concentrated flask, in 40 ℃ water bath spin concentrated to near dry, with ethyl acetate - cyclohexane mixed solvent accurate volume to 1.0
ML, over 0.45 m filter, to be solid phase extraction purification.
7.2 Purification
7.2.1 Vegetable, fruit, grain, walnut, rabbit meat, chicken liver, shrimp, chicken gel chromatography
7.2.1.1 Gel Chromatographic Purification Conditions
A) gel cleaning column. 300 mm × 10 mm (id), Bio Beads S-X3, 60-100 mesh, or equivalent;
B) Mobile phase. cyclohexane-ethyl acetate (1 1);
C) flow rate. 4.7 mL/min;
D) Sample loop. 5 mL;
G) Collection time. 7.5 min ~ 12.5 min.
7.2.1.2 Gel Chromatographic Purification Procedure
10mL of the solution to be purified according to 7.2.1 conditions, the collection of all the collection liquid in the nitrogen blowing tube, in the 35 ℃ water bath blowing to the dry,
Ethyl acetate to 1.0 mL, determined by gas chromatograph, and confirmed by gas chromatography-mass spectrometry.
7.2.2 vinegar, honey
Weigh 2.5 g (accurate to 0.01 g) homogeneous sample, add 5 mL water vortex oscillation mix for 30 s, the solution all through HLB solid phase extraction
Column, the flow rate control in the 1 mL/min, and then 2mL distilled water leaching column, discard the eluent, vacuum drying 2 min, then 4 mL
Alkane-acetone mixed solution, the eluate was collected in a centrifuge tube, dried at 40 ° C under nitrogen and 0.5 mL of ethyl acetate.
7.2.3 tea, onions
Take 0.5 mL of the solution to be purified on the graphitized carbon black and PSA mixed column, eluted with 1.5 mL of ethyl acetate. cyclohexane mixed solvent, flow rate
Control at 1 mL/min, collect all effluent in a centrifuge tube, dry at 40 ° C, 0.5 mL of ethyl acetate.
7.2 Determination
7.3.1 Gas Chromatographic Reference Conditions
A) Column. DB-1301 quartz capillary column, 30m × 0.25 mm (id) × 0.25 m, or equivalent performance;
B) Column temperature. 50 ° C 20 ° C/min.200 ° C 5 ° C/min 260 ° C (10 min);
C) Inlet temperature. 260 ° C;
D) detector temperature. 300 ° C;
E) Carrier gas. nitrogen, purity greater than or equal to 99.999%, column flow 2 mL/min;
F) Injection method. no shunt, 0.75 min after the opening of the diversion valve;
G) Injection volume. 1 μL.
7.3.2 Gas Chromatography - Mass Spectrometry Reference Conditions
A) Column. HP-5MS quartz capillary column, 30m × 0.25 mm (id) × 0.25 m, or equivalent performance;
B) Column temperature. 50 ° C (1 min) 10 ° C/min 280 ° C (10 min);
C) Inlet temperature. 250 ° C;
D) chromatographic - mass spectrometer interface temperature. 280 ° C;
E) ionization mode. EI;
F) Ionization energy. 70 eV;
G) Carrier gas. helium, purity greater than or equal 99.999%, flow rate 1 mL/min;
H) Injection method. no shunt, 0.75 min after the opening of the diversion valve;
I) Injection volume. 1 μL;
J) Measurement method. Select ion monitoring;
K) Select the monitoring ion (m/z). 183,264,265;
L) solvent delay. 5.0 min.
7.3.3 Determination and confirmation of chromatography
According to the content of benzimidium in the sample solution, the standard working solution with similar peak area was selected, the standard working solution and the benzimidium ether phase
The values shall be within the linear range of the instrument. Standard working solution and sample solution volume measurement. Under the above chromatographic conditions, benzyl
The retention time of mite ether was about 22.4 min. The chromatogram of the standard is shown in Appendix A, Appendix A.1.
The standard solution and sample solution are measured according to the conditions of 7.3.1, if the sample solution with the standard solution of the same retention time there is a peak, then
The gas chromatography-mass spectrometry was confirmed.
The results show that the retention time of the chromatogram is consistent with that of the standard sample, and in the sample spectrum after subtracting the background,
The selected ions are present; the abundance of the selected ions is consistent with the relative abundance of the ions associated with the standard sample, and the similarity is allowed
Difference (see Table 1), you can determine the sample for the ciprofloxacin positive detection. Gas Chromatography - Mass Spectrometry Ion Chromatography
And the spectrum shown in Appendix A.2 and Appendix A.3.
Table 1 Maximum allowable deviation of relative ion abundance when qualitative confirmation
Relative abundance (base) 50% 20% to 50% 10% to 20% ≤10%
Allowable relative deviation ± 20% ± 25% ± 30% ± 50%
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 amount of benzimidate residues in the sample.
MA
VCA
Where.
X - the amount of benzimidazole residues in milligrams per kilogram, mg/kg;
A - peak area of acetophenone in sample solution;
Cs - the concentration of benzimidate in standard working fluid, in micrograms per milliliter, g/mL;
V - the final volume of the sample solution, in milliliters, mL;
As - the peak area of benzimidate in standard working solution;
M - the amount of sample represented by the final 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 C 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 D requirements.
10% limit and recovery rate
10.1 Quantitation limits
The limit of quantification of this method is 0.01 mg/kg.
10.2 Recovery rate
When the levels were 0.01 mg/kg, 0.05 mg/kg, 2.0 mg/kg. The addition of benzimidium ether in different substrates is shown in attached
Record
Appendix A
(Informative)
Benzamidate standard chromatogram
Figure A.1 Benzimidate Standard Gas Chromatography
Figure A.2 Benzimidone Gas Chromatography-Mass Spectrometry Ion Chromatography
Figure A.3 Benzomidate standard substance mass spectrometry
Appendix B
(Informative)
The recovery of benzylidene ether in different substrates
Table D.1 Recovery rates of benzylidene ethers in different matrices
Sample added level mg/kg Recovery range (%)
0.01 80.2 to 98.4
0.05 81.6 ~ 97.2
2.0 82.6 ~ 99.1
0.01 84.3 ~ 98.2
0.05 85.2 to 98.2
2.0 87.6 ~ 99.5
asparagus
0.01 82.3 to 101.3
0.05 84.4 to 100.8
2.0 96.1 to 99.7
Green onions
0.01 73.7 to 99.2
0.05 77.0 to 89.8
2.0 81.2 to 102.4
potato
0.01 85.5 to 99.4
0.05 86.2 to 98.6
2.0 85.8 ~ 99.6
honey
0.01 71 to 94.8
0.05 71.8 to 91.4
2.0 81.3 ~ 98.1
Buckwheat
0.01 79 to 96.5
0.05 82.2 to 95.8
2.0 83.1 to 98.4
tea
0.01 78.4 to 104.3
0.05 82.2 to 97.8
2.0 83.2 to 102.1
corn
0.01 81.2 to 102.4
0.05 83.0 to 101.0
2.0 83.6 ~ 99.8
Vinegar
0.01 81 to 96.2
0.05 81.6 ~ 95.0
2.0 83.4 to 98.7
Walnut
0.01 80.3 ~ 97.5
0.05 80.0 to 98.2
2.0 83.8 to 99.1
rabbit
0.01 83.1 to 96.8
0.05 86.2 to 95.6
2.0 85.7 ~ 98.3
Chicken liver
0.01 81.2 to 103.2
0.05 85.0 to 96.5
2.0 81.2 to 90.0
Shrimp
0.01 77.6 ~ 104.1
0.05 79.4 to 99.6
2.0 80.1 to 101.3
chicken
0.01 83.1 to 97.8
0.05 84.4 to 96.2
2.0 84.8 ~ 98.1
Appendix C
(Normative appendix)
Laboratory repeatability requirements
Table C.1 Laboratory repeatability requirements
Measured component content
Mg/kg
Precision
0.001 36
> 0.01
> 1 14
Appendix D
(Normative appendix)
Inter-laboratory reproducibility requirements
Table D.1 Inter-laboratory reproducibility requirements
Measured component content
Mg/kg
Precision
0.001 54
> 0.01
> 1 19
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