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GB 23200.50-2016: Food safety national standard -- Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography -- Mass Spectrometry / Mass Spectrometry
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Food safety national standard -- Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography -- Mass Spectrometry / Mass Spectrometry
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GB 23200.50-2016
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Basic data | Standard ID | GB 23200.50-2016 (GB23200.50-2016) | | Description (Translated English) | Food safety national standard -- Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography -- Mass Spectrometry / Mass Spectrometry | | Sector / Industry | National Standard | | Classification of Chinese Standard | G25 | | Word Count Estimation | 16,110 | | Date of Issue | 2016-12-18 | | Date of Implementation | 2017-06-18 | | Older Standard (superseded by this standard) | SN/T 2561-2010 | | Regulation (derived from) | State Health Commission, Ministry of Agriculture, Food and Drug Administration Notice No. 16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 23200.50-2016: Food safety national standard -- Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography -- Mass Spectrometry / Mass Spectrometry
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Food safety national standard - Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography - Mass Spectrometry/Mass Spectrometry
National Standards of People's Republic of China
GB
Replace SN/T 2561-2010
National standards for food safety
Determination of Pyridine Pesticide Residues in Food
Liquid chromatography - mass spectrometry/mass spectrometry
National food safety standards-
Determination of pyridine pesticides residue in foods
Liquid 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
GB 2320.50-2016
Foreword
This standard replaces SN/T 2561-2010 "Determination of Pyridine Pesticide Residues in Import and Export Food by Liquid Chromatography-Mass Spectrometry".
Compared with SN/T 2561-2010, 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 2561-2010.
National standards for food safety
Determination of Pyridine Pesticide Residues in Food by Liquid Chromatography - Mass Spectrometry/Mass Spectrometry
1 Scope
This standard stipulates that imidacloprid, acetamiprid, imazethapyr, fenfosbenone, pyridylamide, thiazole niacin and fenpropathrin
Determination of seven kinds of pyridine pesticide residues by sample preparation and liquid chromatography - mass spectrometry/mass spectrometry.
This standard applies to rice, wheat, potatoes, spinach, citrus, walnut, tea, pork, fish, liver and milk
Determination and confirmation of the residual residues of insecticides, acetamiprid, imazethapyr, furazolidone, pyridinamide, thiazole niacin and fenpropathrin
It can refer to the implementation of food.
2 normative reference documents
The following documents are indispensable for the application of this document. For dated references, only the date of the note applies
This document. 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
The residual pesticide in the sample was extracted with sodium chloride after salting out the acetonitrile, and the extract was purified by graphitizing carbon black or C18 solid phase extraction column,
Using liquid chromatography - mass spectrometry/mass spectrometry to detect and confirm the external standard method.
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 Acetonitrile (CH3CN). liquid chromatography grade.
4.1.2 Methanol (CH3OH). liquid chromatography grade.
4.1.3 Toluene (C7H8).
4.1.4 acetic acid (CH3COOH).
4.1.5 Sodium chloride (NaCl).
4.1.6 anhydrous sodium sulfate (Na2SO4). 650 ℃ burning 4 h, placed in the dryer to spare.
4.2 solution preparation
4.2.1 0.1% acetic acid solution. take 1mL acetic acid, diluted with water and set to 1000 mL.
4.2.2 methanol - water (1 1, V/V). measure 100 mL of methanol, add 100 mL of water, mix well.
4.2.3 toluene - acetonitrile acidic solution. measure 100 mL of toluene, add 300 mL of acetonitrile and 4 mL of acetic acid, mix well.
4.3 standards
4.3.1 imidacloprid, acetamiprid, imazethapyr, furazolidone, pyridinamide, thiazole niacin and fenpropathrin Standard substance. purity
≥98%, see Appendix A.
4.4 standard solution preparation
4.4.1 standard stock solution. respectively, accurately weighed the appropriate amount of the standard, with methanol prepared into 1.0 mg/mL standard stock solution, 4 ℃
Save under darkness.
4.4.2 Preparation of mixed standard intermediate liquid. Take the above standard stock solution amount, with methanol prepared into 10 g/mL mixed standard solution,
4 ℃ save under the dark.
4.4.3 Preparation of standard working solution. the appropriate amount of mixed standard intermediate solution, with a blank sample matrix prepared into the appropriate concentration of the standard
Working solution, prepared before use.
4.5 Materials
4.5.1 C18 solid phase extraction cartridge. 500 mg, 3 mL or equivalent.
4.5.2 Graphite Carbon Black Column. 500 mg, 6 mL or equivalent.
5 instruments and equipment
5.1 High Performance Liquid Chromatography-Mass Spectrometry/Mass Spectrometer. Distribution Spray Ion Source (ESI).
5.2 Analysis of the balance. the sense of 0.0001 g and 0.01 g.
5.3 Scroll Mixer.
5.4 Centrifuge (maximum speed 10000 r/min).
5.5 nitrogen dryers.
5.6 Rotary Evaporator.
5.7 solid phase extraction device.
5.8 Crusher.
6 Preparation and storage of samples
6.1 Preparation of the sample
During the operation of the sample preparation, the sample should be protected from contamination or changes in the content of the residue.
6.1.1 Citrus, Potato and Spinach
Take a representative sample 500 g, chopped (not available in water), and the sample is processed into a slurry with a mashed machine. Mix well and put it clean
Within the sample container, seal and mark the mark.
6.1.2 Rice, wheat, walnut and tea
Take a representative sample 500 g, crushed with a pulverizer. Mix, fit into a clean container, sealed and marked.
6.1.3 Fish, pork and liver
Take a representative sample 500 g, take the sample by the mashed machine mashed evenly, into the clean sample container, sealed and marked
Remember.
Note. The above sample sampling site according to GB 2763 Appendix A implementation.
6.2 Sample storage
Rice, wheat, walnuts and tea leaves at 0 ℃ 4 ℃; citrus, potatoes, spinach, pork, fish, liver and milk
-18 ℃ below the frozen storage.
7 Analysis steps
7.1 Extraction
7.1.1 Rice, wheat, walnut and tea
For rice, wheat, walnut samples, weigh 2 g sample (accurate to 0.01 g). For tea samples, weigh 1 g sample (fine
Indeed 0.01 g). Place the weighed sample in a 50 mL centrifuge tube, add 5 mL of water to mix for 30 min, add 2 g of sodium chloride and
10 mL of acetonitrile was added to the vortex mixer for 3 min and centrifuged at 5000 r/min for 5 min. The supernatant was transferred to another centrifuge tube.
The extraction process was repeated with 5 mL of acetonitrile and the extracts were combined. Concentrated to about 2 mL at 40 ° C and purged.
7.1.2 Citrus, milk
Weigh 5 g (accurate to 0.01 g) sample in a 50 mL centrifuge tube, add 2 g of sodium chloride and 10 mL of acetonitrile in a vortex mixer
Mixed for 3 min. 5000 r/min centrifugation 5 min, the supernatant transferred to another centrifuge tube. Repeat the above with 5 mL of acetonitrile
Take the process, the combined extract, at 40 ℃ nitrogen blowing concentrated to about 2 mL, to be purified.
7.1.3 Potatoes, spinach, pork, fish and liver
Weigh 2 g (accurate to 0.01 g) sample in a 50 mL centrifuge tube, add 3 mL of water, 2 g of sodium chloride and 10 mL of acetonitrile,
Mix the mixture for 3 min, centrifuge at 5000 r/min for 5 min, transfer the supernatant to another centrifuge tube. Re-use 5 mL of acetonitrile
Repeat the extraction process above, merge the extract. Concentrated to about 2 mL at 40 ° C and purged.
7.2 Purification
7.2.1 Rice, wheat, potatoes, citrus, pork, fish and milk
In the top of the C18 solid phase extraction column into the 1 cm high anhydrous sodium sulfate, first with 5 mL of acetonitrile pre-leaching column, discard the eluent. Then
The resulting extract was passed through the column and eluted with 4 mL of acetonitrile to control the flow rate at 0.5 mL/min. All effluents were collected. At 40 ° C
And the residue was filtered through a 1.0 mL methanol-water solution and passed through a 0.45 μm organic membrane for liquid chromatography-mass spectrometry/mass spectrometry
Determination.
7.2.2 Spinach, liver, walnut and tea
In the top of the graphitized carbon black solid phase extraction column into the 1cm high anhydrous sodium sulfate, first with 10mL toluene - acetonitrile acid solution pre-leaching
Small column, abandoned eluent. The column was then added to the extract and eluted with 25 mL of toluene-acetonitrile acid solution to control the flow rate of 0.5
ML/min, collecting all effluent. In 40 ° C water bath rotation concentrated to about 1 ~ 2mL, and then blown to the near-dry nitrogen flow. With 1.0 mL
Methanol - aqueous solution to dissolve the residue, 0.45 μm of the organic phase microporous membrane, for liquid chromatography - mass spectrometry/mass spectrometry.
7.3 Determination
7.3.1 Liquid Chromatographic Reference Conditions
A) Column. C18 column, 150 4.6 mm (inner diameter), particle size 5 μm or equivalent;
B) mobile phase. gradient elution program in Table 1;
C) Flow rate. 0.5 mL/min;
D) Column temperature. 30 ° C;
E) Injection volume. 10 μL.
Table 1 Mobile phase gradient elution program
time
Min
0.1% acetic acid
Methanol
7.3.2 Mass spectrometry reference conditions
A) ion source. electrospray ion source;
B) scanning mode. positive ions;
C) Detection method. Multiple reaction monitoring (MRM);
D) atomization gas, air curtain gas, auxiliary heating gas, collision gas are high purity nitrogen; before use should adjust the gas flow to mass spectrometry
Sensitivity to meet the test requirements, the reference conditions see Appendix B;
E) Parameters such as spray voltage, de-cluster voltage, and collision energy should be optimized to optimum sensitivity. See Appendix B for reference conditions.
7.3.3 Quantitative determination
According to the sample in the sample content, select the appropriate response value of the standard working solution for analysis. Standard working fluid and sample solution to be tested
The response value of the pyridine pesticide should be within the linear response range of the instrument. If the content exceeds the standard curve range, dilute to the appropriate concentration
Post analysis. The reference retention time of pyridine pesticides under these chromatographic conditions was imidacloprid (8.5 min), acetamiprid (9.0 min)
(11.6 min), paclitaxel (11.9 min), thiazolyl nicotinic acid (12.8 min) and fluorosulfide
(14.0 min). The multi-reaction monitoring chromatogram of the standard solution is shown in Appendix C in Figure C.1.
7.3.4 Qualitative determination
Under the same experimental conditions, the chromatographic peak retention time of the sample in the sample solution was the same as that of the standard working solution, and after subtracting the background
, The relative abundance of each qualitative ion is compared with the abundance of the standard ion, and the deviation
Does not exceed the scope specified in Table 2, you can determine the existence of the corresponding sample in the sample.
Table 2 The maximum allowable deviation of relative ion abundance when qualitative confirmation
Relative ion abundance 50% 20% to 50% 10% to 20% ≤10%
Allowable relative deviation ± 20% ± 25% ± 30% ± 50%
7.4 blank test
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 residual amount of pyridine pesticide in the sample according to formula (1). The calculation result is deductible.
X =
MA
VcA
S
(1)
Where.
The residual amount of pyridine pesticide component in the X - sample, in micrograms per kilogram (μg/kg);
A - the peak area of the pyridine pesticide component in the sample solution;
C - the concentration of pyridine pesticide components in standard working solutions in ng/ml (ng/mL);
V - the final volume of the sample solution in milliliters (mL);
SA - peak area of pyridine pesticide components in standard working solutions;
M - the mass of the 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)
Together with the requirements of Appendix E.
9.2 The ratio of the absolute difference between the two independent determinations obtained under reproducibility and its arithmetic mean (percentage)
Together with the requirements of Appendix F.
10% limit and recovery rate
10.1 Quantitation limits
Quantitative limits for pyridine pesticides in rice, wheat, potatoes, spinach, citrus, walnut, pork, fish, liver and milk
0.005 mg/kg; the limit of quantification of pyridine pesticides in tea was 0.01 mg/kg.
10.2 Recovery rate
Rice, potato, potato, spinach, citrus, walnut, tea, pork, fish, liver and milk.
The data on the level and recovery are shown in Appendix D.
Appendix A
(Informative)
The basic information of the standard substance
Table A.1 Basic information on reference materials
Chinese name English name CAS number Molecular formula Molecular weight
Imidacloprid 138261-41-3 C9H10ClN5O2 255.7
Acetamiprid 160430-64-8 C10H11ClN4 222.7
Imazethapyr 81385-77-5 C15H19N3O3 289.3
Thiazole niacin thiazopyr 117718-60-2 C16H17F5N2O2S 396.4
Flutidone fluridone 59756-60-4 C19H14F3NO 329.3
Pyridinamide boscalid 188425-85-6 C18H12Cl2N2O 343.2
Fisulfuride dithiopyr 97886-45-8 C15H16F5NO2S2 401.4
Appendix B
(Informative)
Reference to mass spectrometry conditions
Reference mass spectrometry conditions.
A) Scanning mode. positive ion scanning;
B) Detection method. Multiple reaction monitoring (MRM);
C) air curtain gas (CUR). 10 Psi;
D) Atomizing gas (GS1). 40 Psi;
E) Auxiliary heated gas (GS2). 50 Psi;
F) collision gas (CAD). 7 Psi;
G) electrospray voltage (IS). 5000 V;
H) ion source temperature (TEM). 450 C;
I) Qualitative ion pair, quantitative ion pair, collision energy, de-cluster voltage, contact chamber inlet and outlet voltage, and collision chamber outlet voltage see
Table B.1.
Table B.1 Multi-reaction monitoring conditions
Compounds
Mother ion
(M/z)
Ion
(M/z)
Dwell time
(Ms)
To cluster voltage
(DP)/V
Collision energy
(CE)/V
Collision room
Inlet voltage
(EP)/V
Collision room
Outlet voltage
(CXP)/V
Imidacloprid 256.0
209.0
175.0.200 46 33 10 6
Acetamidine 223.0
126.0
90.0.200 36 45 10 12
Imazethapyr
290.1
245.6
200 62 26 10 16
177.4.200 62 37 10 15
Fluorothiazone 330.5
309.5
200 124 46 10 15
259.2.200 124 60 10 15
Pyridinamide 343.0
307.2
200 98 27 10 15
140.2.200 98 30 10 6
Thiazole niacin 397.6
377.2
200 94 33 10 11
335.2.200 94 44 10 18
Fluoride Sulfide 402.3
354.0
500 86 26 10 15
272.4 500 98 410 10 15
Note. The ions with "*" are quantitative ions.
1) Non-commercial notices. The parameters listed in Appendix B are performed on the API4000 mass spectrometer, where the test instrument model is listed for reference only and
Do not involve commercial purposes, encourage standard users to try to use different manufacturers or models of equipment.
Appendix C
(Informative)
Multi - reaction monitoring (MRM) chromatograms for standard solutions
Pyridinamides
343.4/307.2
Pyridinamides
343.4/140.2
Acetamidine
223.0/126.0
Acetamidine
223.0/90
Imazethapyr
290.1/245.6
Imitation grass smoke
290.1/177.4
Imidacloprid
256.0/209.0
Imidacloprid
256.0/175.0
Figure C.1 Multi-reaction monitoring (MRM) chromatograms for standard solutions
Fluorothiazone
330.5/259.2 Fluorothiazone
330.5/309.5
Thiazole niacin
397.4/335.2
Thiazole niacin
397.4/377.2
Fenbutetine
402.2/354.0
Fenbutetine
402.2/272.4
Appendix D
(Informative)
The recovery of pyridine pesticides in different sample substrates
Table D.1 Recovery of Pyridine Pesticides in Different Sample Substrates
sample
Matrix
Add to
Level
/ (Mg/kg)
Recovery rate/%
Imidaclopridamphamidine imidazolium niacin fenfluramine ketophenadiazole n -
Rice
0.005 87.8 to 103.0 89.4 to 97.2 73.2 to 90.2 86.2 to 97.6 82.4 to 95.6 64.6 to 83.6 61.2 to 73.4
0.01 97.6 to 105.0 95.5 to 100.5 86.5 to 92.3 89.5 to 103.5 80.8 to 89.9 74.9 to 82.0 60.9 to 78.7
0.025 89.8 to 100.6 92.2 to 99.4 79.0 to 97.6 88.7 to 98.8 86.6 to 99.6 77.8 to 87.5 67.9 to 87.0
potato
0.005 85.0 to 100.2 86.6 to 100.4 65.0 to 86.0 77.0 to 91.2 65.0 to 82.4 69.6 to 79.6 60.6 to 79.6
0.01 88.0 to 96.1 92.9 to 98.5 83.5 to 99.5 79.7 to 87.8 80.5 to 89.0 75.6 to 88.0 65.9 to 76.0
0.025 81.0 to 99.0 82.6 to 100.0 79.6 to 94.7 86.6 to 94.6 79.4 to 88.7 75.2 to 88.1 66.2 to 75.2
Walnut
0.005 82.0 to 95.0 80.2 to 93.2 63.4 to 77.0 79.0 to 95.0 63.6 to 81.0 60.8 to 69.6 66.4 to 74.6
0.01 83.0 to 92.4 89.7 to 96.1 72.5 to 80.3 82.5 to 93.4 78.5 to 89.7 68.0 to 83.4 69.9 to 80.1
0.025 76.4 to 96.6 76.4 to 100.8 69.6 to 77.8 75.8 to 86.2 84.2 to 93.6 64.9 to 76.9 66.2 to 75.2
pork
0.005 76.4 ~ 87.6 75.4 ~ 101.0 60.4 ~ 77.6 75.2 ~ 90.0 61.6 ~ 86.0 60.2 ~ 77.8 60.8 ~ 76.6
0.01 96.7 to 100.3 91.5 to 101.0 70.6 to 101.0 88.5 to 93.0 76.1 to 88.6 71.0 to 81.5 63.4 to 70.1
0.025 87.2 to 99.2 87.1 to 100.0 72.4 to 91.8 87.4 to 95.0 73.4 to 84.3 77.8 to 87.6 63.6 to 68.6
Fish
0.005 91.0 to 102.0 84.8 to 99.4 67.8 to 90.6 79.6 to 91.2 74.2 to 87.4 69.6 to 81.2 64.6 to 79.8
0.01 96.6 to 102.5 94.2 to 102.0 78.9 to 83.4 76.5 to 85.2 80.7 to 89.5 65.5 to 78.9 62.4 to 80.6
0.025 86.5 to 96.6 91.2 to 101.2 80.3 to 91.6 75.8 to 86.2 84.2 to 102.0 66.2 to 83.1 64.9 to 76.9
wheat
0.005 87.0 to 100.2 79.8 to 102.4 81.0 to 93.0 79.4 to 87.8 73.4 to 86.4 73.2 to 83.0 60.2 to 73.0
0.01 89.4 to 104.5 86.8 to 99.9 79.0 to 90.4 85.5 to 95.0 79.5 to 95.5 81.5 to 90.8 62.3 to 78.7
0.025 91.4 to 100.5 86.8 to 99.9 76.6 to 87.9 79.4 to 94.1 71.6 to 87.5 84.0 to 93.0 69.1 to 73.5
Tangerine
0.005 78.0 to 91.2 84.4 to 101.2 75.6 to 95.6 81.2 to 100.0 77.4 to 97.2 75.6 to 94.4 61.4 to 77.8
0.01 87.4 to 97.0 89.0 to 95.6 72.6 to 99.2 81.2 to 100.0 77.2 to 97.8 71.6 to 86.2 79.1 to 85.9
0.025 83.4 to 91.6 84.0 to 99.2 79.6 to 91.2 82.9 to 89.1 77.5 to 87.2 71.8 to 85.8 71.0 to 78.0
milk
0.005 76.4 to 95.2 77.0 to 108.0 69.0 to 93.0 64.6 to 85.0 73.8 to 92.2 65.0 to 82.6 60.0 to 68.8
0.01 81.6 to 97.2 82.4 to 98.0 78.9 to 88.8 83.6 to 91.5 68.4 to 88.4 67.2 to 84.2 63.2 to 81.2
0.025 82.4 to 94.0 81.9 to 92.8 80.8 to 97.2 85.8 to 106.8 78.2 to 87.6 71.6 to 87.6 67.2 to 77.7
spinach
0.005 72.4 to 88.4 76.6 to 97.8 68.6 to 80.4 77.2 to 91.4 64.6 to 82.4 63.6 to 77.6 60.4 to 69.0
0.01 81.4 to 92.0 87.6 to 98.5 74.2 to 87.6 79.8 to 88.8 77.8 to 84.5 67.9 to 82.3 61.3 to 80.1
0.025 76.6 to 98.0 78.6 to 89.2 78.6 to 87.0 82.8 to 91.6 77.4 to 92.8 67.4 to 79.5 64.0 to 76.6
Liver
0.005 75.0 to 89.0 70.6 to 89.2 62.8 to 74.0 72.6 to 80.8 74.6 to 82.6 61.6 to 71.8 64.4 to 75.6
0.01 72.7 to 84.6 84.4 to 91.5 60.0 to 80.4 82.5 to 92.9 71.2 to 81.4 72.3 to 80.1 60.3 to 73.8
0.025 72.8 ~ 90.4 78.8 ~ 99.9 72.9 ~ 76.9 77.7 ~ 95.0 74.4 ~ 85.6 68.5 ~ 82.3 63.0 ~ 76.4
tea
0.01 70.0 to 82.0 83.2 to 94.1 66.6 to 77.8 79.6 to 94.6 68.4 to 80.2 62.6 to 78.2 62.0 to 78.8
0.02 78.9 to 94.5 76.8 to 94.5 71.7 to 84.8 78.6 to 101.5 67.8 to 91.0 73.5 to 86.5 62.5 to 84.5
0.05 70.2 to 85.2 78.8 to 93.2 68.0 to 75.8 76.5 to 91.9 65.4 to 73.4 71.1 to 79.8 70.2 to 85.2
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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