GB 5009.291-2023 English PDF

GB 5009.291-2023: National food safety standard - Determination of chlorate and perchlorate in foods
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GB 5009.291-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards - Determination of chlorate
and perchlorate in food
ISSUED ON. SEPTEMBER 6, 2023
IMPLEMENTED ON. MARCH 6, 2024
Issued by. National Health Commission of the PRC;
State Administration for Market Regulation.
Table of Contents
1 Scope... 3
2 Principle... 3
3 Reagents and materials... 3
4 Apparatus... 6
5 Analytical procedure... 6
6 Expression of analysis results... 9
7 Precision... 10
8 Other... 10
Appendix A Reference conditions for liquid chromatography-tandem mass
spectrometry... 11
Appendix B Multiple reaction monitoring (MRM) diagrams of chlorate and
perchlorate standards... 13
National food safety standards - Determination of chlorate
and perchlorate in food
1 Scope
This Standard specifies the liquid chromatography-tandem mass spectrometry method
for the determination of chlorate and perchlorate in food.
This Standard applies to the determination of chlorate and perchlorate in vegetables and
their products, fruits and their products, cereals and their products, meat and meat
products, aquatic products, eggs and egg products, milk and dairy products, condiments,
beverages, infant formula foods, infant supplementary foods, and tea.
2 Principle
Use an acidified acetonitrile-water mixed solution to extract the chlorate and
perchlorate in the specimen; use a solid-phase extraction column to purify; use high-
performance liquid chromatography to separate; use tandem mass spectrometry to test;
use the isotope internal standard method to quantify.
3 Reagents and materials
Unless otherwise stated, all reagents in this method are chromatographic grade; the
water is grade 1 water specified in GB/T 6682.
3.1 Reagents
3.1.1 Acetonitrile (CH3CN).
3.1.2 Methanol (CH3OH).
3.1.3 Formic acid (HCOOH).
3.2 Reagent preparation
3.2.1 0.1% formic acid aqueous solution. Take 1 mL of formic acid; use water to dilute
to 1000 mL and mix well.
3.2.2 Acetonitrile-formic acid aqueous solution. Take 60 mL of acetonitrile; use 0.1%
formic acid aqueous solution to adjust the volume to 100 mL; mix well.
3.3 Standards
3.3.1 Sodium chlorate (NaClO3, CAS number. 7775-09-9). Purity is >99%; or a
standard certified by the state and granted the reference material certificate.
3.3.2 Sodium perchlorate (NaClO4, CAS number. 7601-89-0). Purity is >99%; or a
standard certified by the state and granted the reference material certificate.
3.3.3 18O3-chlorate (chlorate isotope internal standard). 100 μg/mL (calculated as 18O3-
chlorate).
3.3.4 18O4-perchlorate (perchlorate isotope internal standard). 100 μg/mL (calculated as
18O4-perchlorate).
3.4 Preparation of standard solutions
3.4.1 Chlorate standard stock solution (1.0 mg/mL, calculated as chlorate). Accurately
weigh 1.2801 g of sodium chlorate; use water to dissolve and dilute to 1000 mL; mix
well. Store it at 2 °C~8 °C. The validity period is 1 year.
3.4.2 Perchlorate standard stock solution (1.0 mg/mL, calculated as perchlorate).
Accurately weigh 1.2300 g of sodium perchlorate; use water to dissolve and dilute to
1000 mL; mix well. Store it at 2 °C~8 °C. The validity period is 1 year.
Note. The standard stock solutions of chlorate and perchlorate can use standards certified by the
state and granted the reference material certificate.
3.4.3 Perchlorate standard stock intermediate solution (100 μg/mL). Accurately pipette
10.0 mL of perchlorate standard stock solution; place it in a 100 mL volumetric flask;
use water to dilute and make up to volume; mix well; transfer the solution to a reagent
bottle. Store it at 2 °C~8 °C. The validity period is 1 year.
3.4.4 Chlorate and perchlorate mixed standard intermediate solution. Respectively
pipette 1.00 mL of chlorate standard stock solution AND 1.00 mL of perchlorate
standard stock intermediate solution; place them in the same 100 mL volumetric flask.
Use water to dilute and make up to volume; shake well; prepare a mixed standard
intermediate solution with mass concentrations of chlorate and perchlorate of 10 μg/mL
and 1.0 μg/mL respectively; transfer the solution to a reagent bottle. Store it at
2 °C~8 °C. The validity period is 3 months.
3.4.5 Chlorate and perchlorate mixed standard use solution Ⅰ. Pipette 1.00 mL of
chlorate and perchlorate mixed standard intermediate solution; place it in a 10 mL
volumetric flask. Use water to dilute and make up to volume; shake well; prepare a
mixed standard use solution with mass concentrations of chlorate and perchlorate of
1000 ng/mL and 100 ng/mL respectively; transfer the solution to a reagent bottle. Store
it at 2 °C~8 °C. The validity period is 1 month.
3.4.6 Chlorate and perchlorate mixed standard use solution II. Pipette 1.00 mL of
chlorate and perchlorate mixed standard use solution I; place it in a 10 mL volumetric
flask. Use water to dilute and make up to volume; shake well; prepare a mixed standard
use solution with mass concentrations of chlorate and perchlorate of 100 ng/mL and 10
ng/mL respectively; transfer the solution to a reagent bottle. Store it at 2 °C~8 °C. The
validity period is 1 month.
3.4.7 Chlorate and perchlorate isotope internal-standard mixed standard intermediate
solution. Pipette 0.50 mL of chlorate isotope internal standard AND 50 μL of
perchlorate isotope internal standard; place them in the same 10 mL volumetric flask.
Use water to dilute and make up to volume; shake well; prepare a mixed standard
intermediate solution with mass concentrations of chlorate internal standard and
perchlorate internal standard of 5.0 μg/mL and 0.50 μg/mL, respectively. Store it at
2 °C~8 °C. The validity period is 3 months.
3.4.8 Chlorate and perchlorate isotope internal-standard mixed standard use solution.
Pipette 2.00 mL of chlorate and perchlorate isotope internal-standard mixed standard
intermediate solution; place it in a 10 mL volumetric flask. Use water to dilute and make
up to volume; shake well; prepare a mixed standard use solution with mass
concentrations of chlorate internal standard and perchlorate internal standard of 1.0
μg/mL and 0.10 μg/mL, respectively. Store it at 2 °C~8 °C. The validity period is 3
months.
3.4.9 Chlorate and perchlorate standard series solutions. Respectively pipette 0 μL, 250
μL, and 500 μL of chlorate and perchlorate mixed standard solution II; 100 μL, 500 μL,
1000 μL, and 5000 μL of chlorate and perchlorate mixed standard use solution I; AND
200 μL of isotope internal-standard mixed standard use solution. Use water to dilute
and adjust the volume to 10 mL. The mass concentrations of chlorate are 0 ng/mL, 2.5
ng/mL, 5.0 ng/mL, 10 ng/mL, 50 ng/mL, 100 ng/mL, and 500 ng/mL, respectively. The
mass concentrations of perchlorate are 0 ng/mL, 0.25 ng/mL, 0.50 ng/mL, 1.0 ng/mL,
5.0 ng/mL, 10 ng/mL, and 50 ng/mL, respectively. The mass concentration of the
chlorate isotope internal standard solution in the standard working solution is 20 ng/mL;
the concentration of the perchlorate isotope internal standard solution is 2.0 ng/mL.
Prepare at the time of use.
3.5 Materials
3.5.1 Solid-phase extraction column. Graphitized carbon black column (500 mg, 6 mL)
or one with equivalent performance. Before use, use 6 mL of methanol for activation;
use 6 mL of acetonitrile-formic acid aqueous solution to elute and balance. Before
loading the sample, drain the remaining solution in the column.
3.5.2 Ag/H column (1 g, 2.5 mL) or one with equivalent performance. Before use, use
10 mL of water to rinse; let stand and activate for 30 min.
Weigh 1 g of specimen (accurate to 0.01 g); add 20.0 μL of chlorate and perchlorate
isotope internal-standard mixed standard use solution; vortex for 10 s; use a liquid
chromatograph-tandem mass spectrometer for determination.
5.2.2 Other samples
For solid samples or condiments, weigh 1 g (accurate to 0.01 g). For liquid or semi-
solid samples (except condiments), weigh 2 g (accurate to 0.01 g). Place in a 50 mL
centrifuge tube; add 40.0 μL of chlorate and perchlorate isotope internal-standard mixed
standard intermediate solution; mix by shaking and let stand for 30 min. Add 4.0 mL of
0.1% formic acid aqueous solution; shake and mix after plugging; vortex and mix for
30 s; ultrasonic extraction for 15 min; add 6.0 mL of acetonitrile; ultrasonic extraction
for 30 min; centrifuge at 10000 r/min for 5 min; take the supernatant for purification.
5.3 Specimen purification
5.3.1 High-salt samples (soy sauce, oyster sauce, spiced salt, etc.)
Pipette 2.0 mL of the supernatant through the graphitized carbon black solid-phase
extraction column; discard the effluent; then pipette 5.0 mL of the supernatant through
the graphitized carbon black solid-phase extraction column; use a centrifuge tube to
collect the effluent. Then use an Ag/H cartridge to purify; discard the first 3 mL; collect
the subsequent effluent; pass it through a regenerated cellulose filter membrane; use a
liquid chromatograph-tandem mass spectrometer for determination.
5.3.2 Other samples
Pipette 2.0 mL of the supernatant through the graphitized carbon black solid-phase
extraction column; discard the effluent. Then pipette 2.0 mL of the supernatant through
the graphitized carbon black solid-phase extraction column; pass the effluent through
the regenerated cellulose filter membrane; use a liquid chromatograph-tandem mass
spectrometer for determination.
5.4 Reference conditions for liquid chromatography-tandem mass spectrometry
5.4.1 Liquid chromatography conditions are as follows.
a) Chromatographic column. Pentafluorophenyl column with positive charge on the
packing surface (column length is 100 mm; inner diameter is 2.1 mm; packing
particle size is 1.7 μm); or one with equivalent performance.
b) Mobile phase. Phase A is acetonitrile; phase B is 0.1% formic acid aqueous
solution. The gradient elution procedure is shown in Table A.1 in Appendix A.
c) Flow rate. 0.3 mL/min.
d) Column temperature. 35 °C.
e) Injection volume. 3 μL.
5.4.2 Mass spectrometry conditions are as follows.
a) Ionization mode. Electrospray ionization negative ion mode (ESI-).
b) Scanning method. Multiple reaction monitoring (MRM) scan.
c) Mass spectrometry tuning parameters AND qualitative and quantitative ions are
shown in Table A.2 in Appendix A.
d) The multiple reaction monitoring (MRM) ion channel diagrams of chlorate and
perchlorate are shown in Figure B.1 in Appendix B.
5.5 Preparation of standard curve
Use liquid chromatography-tandem mass spectrometry to determine the chlorate and
perchlorate standard series solutions; obtain the corresponding peak area. Use the
concentration of chlorate and perchlorate in the standard series working solution as the
abscissa; use the peak area ratio as the ordinate; draw a standard curve.
5.6 Qualitative determination
Select two groups of ions for the component to be measured. Each group of ions
contains a parent ion and a product ion. Under the same test conditions, the retention
time of the substance to be tested in the sample AND the corresponding retention time
in the standard solution deviate within 2.5%. The signal-to-noise ratio of the monitored
ion pairs must be greater than or equal to 3 (S/N≥3). And the relative abundance of the
qualitative ion of the measured component in the sample IS compared with the relative
abundance of the corresponding qualitative ion in the standard solution with a similar
concentration. If the deviation does not exceed the range specified in Table 1, it can be
determined that there is a corresponding analyte in the sample.
Table 1 -- Maximum allowable deviation of relative ion abundance in qualitative
confirmation
5.7 Quantitative determination
The specimen solution obtained by the treatment is analyzed by liquid chromatograph-
tandem mass spectrometer; to obtain the ratio OF the peak area of chlorate and
perchlorate TO the peak area of the corresponding isotope internal standard. According
to the standard curve, obtain the contents of chlorate and perchlorate in the liquid to be
tested.
Maximum allowable deviation/%
Relative ion abundance (K)/%
GB 5009.291-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards - Determination of chlorate
and perchlorate in food
ISSUED ON. SEPTEMBER 6, 2023
IMPLEMENTED ON. MARCH 6, 2024
Issued by. National Health Commission of the PRC;
State Administration for Market Regulation.
Table of Contents
1 Scope... 3
2 Principle... 3
3 Reagents and materials... 3
4 Apparatus... 6
5 Analytical procedure... 6
6 Expression of analysis results... 9
7 Precision... 10
8 Other... 10
Appendix A Reference conditions for liquid chromatography-tandem mass
spectrometry... 11
Appendix B Multiple reaction monitoring (MRM) diagrams of chlorate and
perchlorate standards... 13
National food safety standards - Determination of chlorate
and perchlorate in food
1 Scope
This Standard specifies the liquid chromatography-tandem mass spectrometry method
for the determination of chlorate and perchlorate in food.
This Standard applies to the determination of chlorate and perchlorate in vegetables and
their products, fruits and their products, cereals and their products, meat and meat
products, aquatic products, eggs and egg products, milk and dairy products, condiments,
beverages, infant formula foods, infant supplementary foods, and tea.
2 Principle
Use an acidified acetonitrile-water mixed solution to extract the chlorate and
perchlorate in the specimen; use a solid-phase extraction column to purify; use high-
performance liquid chromatography to separate; use tandem mass spectrometry to test;
use the isotope internal standard method to quantify.
3 Reagents and materials
Unless otherwise stated, all reagents in this method are chromatographic grade; the
water is grade 1 water specified in GB/T 6682.
3.1 Reagents
3.1.1 Acetonitrile (CH3CN).
3.1.2 Methanol (CH3OH).
3.1.3 Form...