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GB 5009.82-2016 English PDF (GB5009.82-2016)

GB 5009.82-2016 English PDF (GB5009.82-2016)

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GB 5009.82-2016: Determination of retinol and tocopherol in foods

This standard specifies the determination of vitamin A, vitamin E and vitamin D in food. The first method of this standard is applicable to the determination of vitamin A and vitamin E in food.
GB 5009.82-2016
GB
NATIONAL STANDARD
OF THE PEOPLE REPUBLIC OF CHINA
National Food Safety Standard
Determination of vitamin A, D, E in food
ISSUED ON. DECEMBER 23, 2016
IMPLEMENTED ON. JUNE 23, 2017
Issued by. National Health and Family Planning Commission of the
People's Republic of China;
China Food and Drug Administration.
Table of contents
Foreword ... 4
1 Scope ... 5
2 Principles ... 5
3 Reagents and materials ... 5
4 Instruments and equipment ... 8
5 Analytical procedures ... 8
6 Expression of analytical results ... 11
7 Precision... 12
8 Others ... 12
9 Principles ... 12
10 Reagents and materials... 12
11 Instruments and equipment ... 14
12 Analytical procedures ... 15
13 Expression of results ... 17
14 Precision ... 18
15 Others ... 18
16 Principles ... 18
17 Reagents and materials... 18
18 Instruments and equipment ... 21
19 Analytical procedures ... 21
20 Presentation of analytical results ... 25
21 Precision ... 25
22 Others ... 26
23 Principles ... 26
24 Reagents and materials... 26
25 Instruments and equipment ... 29
26 Analytical procedures ... 29
27 Presentation of analytical results ... 33
28 Precision ... 33
29 Others ... 33
Appendix A ... 34
Appendix B Vitamin A, D, E standard solution concentration calibration method ... 36
Appendix C Chromatogram ... 38
Foreword
This standard replaces GB/T 5009.82-2003 ?€?Determination of retinol and
tocopherol in food?€?, GB 5413.9-2010 ?€?National food safety standard -
Determination of vitamin A, D, E in foods for infants and young children, milk and milk products?€?, GB/T 9695.26-2008 ?€?Meat and meat products -
Determination of vitamin A content?€?, GB/T 9695.30-2008 ?€?Meat and meat
products - Determination of vitamin E content?€?, and NY/T 1598-2008
?€?Determination of tocopherol content in edible vegetable oils - High
performance liquid chromatography?€?.
As compared with GB/T 5009.82-2003, the main changes of this standard are as follows.
- CHANGE the standard name into ?€?National Food Safety Standard
Determination of vitamin A, D, E in food";
- ADD the ?€?Determination of vitamin E in food - Normal phase high
performance liquid chromatography?€?;
- ADD the ?€?Determination of vitamin D in food - Liquid chromatography - Tandem mass spectrometry?€?;
- ADD the ?€?Determination of vitamin D in food - High performance liquid chromatography";
- MODIFY the ?€?Determination of vitamin A and vitamin E in food - Reversed- phase high performance liquid chromatography?€?;
- MODIFY the reverse phase chromatography separation conditions of
vitamin E isomers, which can simultaneously separate and determine 4
tocopherol isomers;
- DELETE the benzopyrene internal standard quantitative method; CHANGE
it to using the external standard method for quantitative;
- DELETE the ?€?colorimetry?€? to determine vitamin A.
National Food Safety Standard
Determination of vitamin A, D, E in food
1 Scope
This standard specifies the determination of vitamin A, vitamin E and vitamin D in food.
The first method of this standard is applicable to the determination of vitamin A and vitamin E in food.
The second method of this standard is applicable to the determination of vitamin E in edible oil, nuts, beans, pepper powder and other foods.
The third method of this standard is applicable to the determination of vitamin D2 and vitamin D3 in food.
The fourth method of this standard is applicable to the determination of vitamin D2 or vitamin D3 in formulated foods.
Method 1 Determination of vitamin A and vitamin E in food - Reversed-
phase high performance liquid chromatography
2 Principles
The vitamin A and vitamin E in the test specimen are saponified (digested by amylase if containing starch), extracted, purified and concentrated, then separated by C30 or PFP reversed-phase liquid chromatography column,
detected by ultraviolet detector or fluorescence detector, AND quantitative by external standard method.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are of analytical grade pure AND water is the primary water as specified in GB/T 6682.
3.1 Reagents
3.1.1 Absolute ethanol (C2H5OH). it shall not contain aldehydes after inspection, AND the inspection method is as shown in A.1.
3.3.2.3 ??-tocopherol (C28H48O2, CAS No.. 54-28-4). purity ??? 95%, OR the standard substance as certified by the state and awarded with the standard substance certificate.
3.3.2.4 ??-Tocopherol (C27H46O2, CAS No.. 119-13-1). purity ??? 95%, OR the standard substance as certified by the state and awarded with the standard substance certificate.
3.4 Standard solution preparation
3.4.1 Vitamin A standard stock solution (0.500 mg/mL). accurately WEIGH 25.0 mg vitamin A standard substance; USE absolute ethanol to dissolve it;
TRANSFER it into a 50 mL volumetric flask; MAKE the volume reach to the mark; at this time the solution concentration is about 0.500 mg/mL. TRANSFER the solution into a brown reagent bottle; SEAL it and PRESERVE it at -20 ??C in the dark; AND the valid period is 1 month. Before use, INCREASE the
temperature of the solution back to 20 ??C; and CONDUCT concentration
correction (as for the correction method, SEE Appendix B).
3.4.2 Vitamin E standard stock solution (1.00 mg/mL). respectively and
accurately WEIGH 50.0 mg of each of ??-tocopherol, ??-tocopherol ??-tocopherol and ??-Tocopherol; USE absolute ethanol to dissolve it; TRANSFER it into a 50 mL volumetric flask; MAKE the volume reach to the mark; at this time the solution concentration is about 1.00 mg/mL. TRANSFER the solution into a brown reagent bottle; SEAL it and PRESERVE it at -20 ??C; AND the valid period is 6 month. Before use, INCREASE the temperature of the solution back to 20 ??C; and CONDUCT concentration correction (as for the correction method, SEE Appendix B).
3.4.3 Intermediate solution of vitamin A and vitamin E mixed standard solution. Accurately PIPETTE 1.00 mL of vitamin A standard stock solution and 5.00 mL of vitamin E standard stock solution into one 50 mL volumetric flask; USE methanol make it reach to the mark; at this time, the vitamin A concentration of the solution is 10.0 ??g/mL, AND each tocopherol concentration of the vitamin E is 100 ??g/mL. PRESERVE it at -20 ??C in dark; AND the valid period is half a month.
3.4.4 Vitamin A and vitamin E standard series working solution. respectively and accurately PIPETTE 0.20 mL, 0.50 mL, 1.00 mL, 2.00 mL, 4.00 mL, 6.00 mL of the intermediate solution of vitamin A and vitamin E mixed standard solution into a 10 mL brown volumetric flask; USE methanol to make the volume reach to the mark; the vitamin A concentration of this standard series is 0.20 ??g/mL, 0.50 ??g/mL, 1.00 ??g/mL, 2.00 ??g/mL, 4.00 ??g/mL, and 6.00 ??g/mL, AND the vitamin E concentration is 2.00 ??g/mL, 5.00 ??g/mL, 10.0 ??g/mL, 20.0 ??g/mL, 40.0 ??g/mL, and 60.0 ??g/mL. PREPARE it before use.
Note. saponification period is generally 30 min; after the saponification solution is cooled down, if there is floating oil on the surface, it shall a...

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