GB 5009.22-2016 PDF English
US$245.00 · In stock · Download in 9 secondsGB 5009.22-2016: National food safety standard - B-group and G-group aflatoxins in foods Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedureStatus: Valid GB 5009.22: Evolution and historical versions
| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.22-2016 | English | 245 |
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National food safety standard - B-group and G-group aflatoxins in foods
| Valid |
| GB/T 5009.22-2003 | English | 479 |
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Determination of aflatoxin B1 in foods
| Obsolete |
| GB/T 5009.22-1996 | English | 359 |
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Method for determination of aflatoxin B1 in foods
| Obsolete |
| GB 5009.22-1985 | English | 239 |
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2 days
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Method for determination of aflatoxin B1 in foods
| Obsolete |
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GB 5009.22-2016: National food safety standard - B-group and G-group aflatoxins in foods---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GB5009.22-2016
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard - Determination of B-
group and G-group Aflatoxins in Foods
Issued on. DECEMBER 23, 2016
Implemented on. JUNE 23, 2017
Issued by. National Health and Family Planning Commission of the PRC;
China Food and Drug Administration.
Table of Contents
Foreword... 4
1 Scope... 5
2 Principle... 6
3 Reagents and materials... 6
4 Instruments and equipment... 8
5 Analytical procedures... 9
6 Expression of analytical results... 15
7 Precision... 15
8 Others... 16
9 Principle... 16
10 Reagents and materials... 16
11 Instruments and equipment... 18
12 Analytical procedures... 19
13 Expression of analytical results... 21
14 Precision... 22
15 Others... 22
16 Principle... 23
17 Reagents and materials... 23
18 Instruments and equipment... 25
19 Analytical procedures... 27
20 Expression of analytical results... 31
21 Precision... 32
22 Others... 32
23 Principle... 33
24 Reagents and materials... 33
25 Instruments and equipment... 33
26 Analytical procedures... 34
27 Expression of analytical results... 34
28 Precision... 35
29 Others... 35
30 Principle... 36
31 Reagents and materials... 36
32 Instruments and equipment... 38
33 Analytical steps... 38
34 Precision... 46
35 Others... 46
Appendix A Calibration method of standard concentration of AFT B1, AFT B2,
AFT G1 and AFT G2... 47
Appendix B Method of verification of immunoaffinity column... 49
Appendix C Tandem mass spectrometry... 50
Appendix D Liquid chromatogram... 55
Appendix E Determination method of quality of enzyme-linked immunosorbent
kit... 59
1 Scope
This standard specifies the method for determination of aflatoxin B1, aflatoxin
B2, aflatoxin G1, aflatoxin G2 (hereinafter referred to as AFT B1, AFT B2, AFT G1
and AFT G2) in food.
The first method of this standard is the isotope dilution liquid chromatography-
tandem mass spectrometry, which is suitable for the determination of AFT B1,
AFT B2, AFT G1 and AFT G2 in cereals and their products, beans and their
products, nuts and seeds, fats and their products, seasonings, infant formula
and infant complementary foods.
The second method of this standard is the high-performance liquid
chromatography-pre-column derivatization, which is suitable for the
determination of AFT B1, AFT B2, AFT G1 and AFT G2 in cereals and their
products, beans and their products, nuts and seeds, fats and their products,
seasonings, infant formula and infant complementary foods.
The third method of this standard is the high-performance liquid
chromatography-post-column derivatization, which is suitable for the
determination of AFT B1, AFT B2, AFT G1 and AFT G2 in cereals and their
products, beans and their products, nuts and seeds, fats and their products,
seasonings, infant formula and infant complementary foods.
The fourth method of this standard is the enzyme-linked immunoadsorption
screening method, which is suitable for the determination of AFT B1 in cereals
and their products, beans and their products, nuts and seeds, fats and their
products, seasonings, infant formula and infant complementary foods.
The fifth method of this standard is the thin-layer chromatography, which is
suitable for the determination of AFT B1 in cereals and their products, beans
and their products, nuts and seeds, fats and their products, and seasonings.
Method I. Isotope dilution liquid chromatography-
tandem mass spectrometry
2 Principle
The aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2 in the specimen are
extracted by acetonitrile-water solution or methanol-water solution. After the
extract is diluted by the phosphate buffer solution which contains 1% Triton X-
100 (or Tween-20) (if necessary, it is initially purified by the aflatoxin solid-phase
purification column), through purification and enrichment by immunoaffinity
column, the purification liquid is concentrated, constant-volume, filtered,
separated by liquid chromatography, tested by tandem mass spectrometry, then
subjected to quantification by isotope internal standard method.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are of analytical
grade, the water is the grade I water as specified in GB/T 6682.
3.1 Reagents
3.2 Preparation of reagents
3.3 Standard substance
3.3.1 AFT B1 standard substance (C17H12O6, CAS. 1162-65-8). Purity ≥ 98%, or
3.4 Preparation of standard solution
4 Instruments and equipment
4.1 Homogenizer.
4.2 High-speed pulverizer.
4.3 Tissue masher.
4.4 Ultrasonic / vortex oscillator or shaker.
4.5 Balance. Sensitivity is 0.01 g and 0.00001 g.
4.6 Vortex mixer.
4.9 Glass-fiber filter paper. Fast, high-load, which retains 1.6 μm particles in
liquid.
4.13 Liquid chromatography column.
4.14 Immunoaffinity column. Capacity of AFT B1 column ≥ 200 ng, recovery rate
of AFT B1 column ≥ 80%, cross-reaction rate of AFT G2 ≥ 80% (see Appendix B
for verification method).
4.15 Aflatoxin-specific solid-phase extraction purification column or functionally
equivalent solid-phase extraction column (hereinafter referred to as purification
column). which is used for the determination of samples of complex matrix.
4.18 pH meter.
5 Analytical procedures
The use of immunoaffinity columns from different manufacturers may slightly
differ in sample loading, rinsing and elution operations, so it shall be performed
in accordance with the operation instructions as provided by the manufacturer.
Caution.
5.1 Preparation of sample
5.1.1 Liquid samples (vegetable oil, soy sauce, vinegar, etc.)
The sampling amount shall be greater than 1 L. For packaging samples such
as bags and bottles, it shall collect at least 3 packages (same batch or number).
All liquid samples shall be mixed by a homogenizer in a container, any 100 g
(mL) of sample is taken for testing.
5.1.2 Solid samples (cereals and their products, nuts and seeds, cereal
supplements for infants, etc.)
The sampling amount shall be greater than 1 kg. It shall be crushed by high-
speed pulverizer and sieved to make the particle size less than the 2 mm
aperture test sieve. It is evenly mixed and then reduced to 100 g, stored in the
sample bottle, sealed for preservation, to prepare for testing.
5.1.3 Semi-fluid (fermented bean curd, fermented soybean, etc.)
The sampling amount shall be greater than 1 kg (L). For packaging samples
such as bags and bottles, it shall collect at least 3 packages (same batch or
number). Then it is crushed and mixed uniformly by the tissue crusher, stored
in sample bottle, sealed for preservation, to prepare for testing.
5.2 Extraction of sample
5.2.1 Liquid sample
5.2.2 Solid sample
5.2.2.1 General solid samples
WEIGH 5 g of specimen (accurate to 0.01 g) into a 50 mL centrifuge tube; ADD
100 μL of isotope internal standard working solution; OSCILLATE to mix it; LET
it be standing for 30 min. ADD 20.0 mL of acetonitrile-water solution (84 + 16)
or methanol-water solution (70 + 30); VORTEX to mix it uniformly; PLACE it in
the ultrasonic/vortex oscillator or shaker to oscillate it for 20 min (or
homogenized by homogenizer for 3 min)
5.2.2.2 Infant formula and infant complementary foods
WEIGH 5 g of specimen (accurate to 0.01 g) into a 50 mL centrifuge tube; ADD
100 μL of isotope internal standard working solution; OSCILLATE to mix it; LET
it be standing for 30 min. ADD 20.0 mL of acetonitrile-water solution (50 + 50)
or methanol-water solution (70 + 30); VORTEX to mix it uniformly; PLACE it in
the ultrasonic/vortex oscillator or shaker to oscillate it for 20 min (or
homogenized by homogenizer for 3 min).
5.2.3 Semi-fluid sample
WEIGH 5 g of specimen (accurate to 0.01 g) into a 50 mL centrifuge tube; ADD
100 μL of isotope internal standard working solution; OSCILLATE to mix it; LET
it be standing for 30 min. ADD 20.0 mL of acetonitrile-water solution (84 + 16)
or methanol-water solution (70 + 30); PLACE it in the ultrasonic/vortex oscillator
or shaker to oscillate it for 20 min (or homogenized by homogenizer for 3 min).
CENTRIFUGE it at 6000 r/min for 10 min (or otherwise homogenized and then
filtered by glass-fiber filter paper), TAKE the supernatant to prepare for use.
5.3 Purification of sample
5.3.1 Purification of immunoaffinity column
5.3.2 Simultaneous use of aflatoxin solid-phase purification column and
immunoaffinity column (for complex substrates such as Chinese red
pepper, black pepper and chili pepper)
5.5 Reference conditions for mass spectrometry
The reference conditions for mass spectrometry are listed below.
5.6 Qualitative determination
The retention time of the chromatographic peak of the target compound in the
specimen is compared with the retention time of the corresponding standard
chromatographic peak, the variation range shall be within ±2.5%.
The mass spectrometric ions of each compound must appear, including at least
one parent ion and two daughter ions. Meanwhile for the same batch under
testing and for the same compound, the allowable deviation between the
relative abundance ratio of the two daughter ions of the target compound in the
sample and that of the standard solution of equivalent concentration shall not
exceed the range as specified in Table 3.
5.7 Production of standard curve
Under the analytical conditions of liquid chromatography-tandem mass
spectrometry of 5.4 and 5.5, MAKE the standard series solution subject to
injection testing in the order from low to high concentration; USE the area ratio
of the AFT B1, AFT B2, AFT G1 and AFT G2 chromatogram peaks to each
corresponding internal standard chromatogram peak as well as the
concentration to make drawing, to obtain the standard curve regression
equation, the linear correlation coefficient of which is greater than 0.99.
5.8 Determination of sample solution
TAKE the sample solution to be tested which is processed in 5.3; USE the
internal standard method to calculate the mass concentration of the target
substance in the tested solution. Follow the provisions of clause 6 to calculate
the content of the tested substance in the sample. The response value in the
5.9 Blank test
DO not weigh the specimen; PERFORM the blank test in accordance with the
steps of 5.2 and 5.3.It shall be confirmed that it does not contain substances
that interfere with the component to be tested.
6 Expression of analytical results
The residual amount of AFT B1, AFT B2, AFT G1 and AFT G2 in the specimen is
calculated in accordance with formula (1).
7 Precision
The absolute difference between two independent determinations obtained
under repeatability conditions shall not exceed 20% of the arithmetic mean.
8 Others
When 5 g of the sample is weighed, the detection limit of AFT B1 is 0.03 μg/kg,
the detection limit of AFT B2 is 0.03 μg/kg, the detection limit of AFT G1 is 0.03
μg/kg, the detection limit of AFT G2 is 0.03 μg/kg; the limit of quantification of
AFT B1 is 0.1 μg/kg, the limit of quantification of AFT B2 is 0.1 μg/kg, the limit of
quantification of AFT G1 is 0.1 μg/kg, the limit of quantification of AFT G2 is 0.1
μg/kg.
9 Principle
The aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2 in the specimen are
extracted by a mixed solution of acetonitrile-water solution or methanol-water
solution, the extract is purified by aflatoxin solid-phase purification column to
remove the interfering substances such as fats, proteins, pigments and
carbohydrates.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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