GB 5009.82-2016 PDF English
US$135.00 · In stock · Download in 9 secondsGB 5009.82-2016: National Food Safety Standard Determination of vitamin A, D, E in food Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedureStatus: Valid GB 5009.82: Evolution and historical versions
| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.82-2016 | English | 135 |
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National Food Safety Standard Determination of vitamin A, D, E in food
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| GB/T 5009.82-2003 | English | 70 |
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Determination of retinol and tocopherol in foods
| Obsolete |
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GB 5009.82-2016: National Food Safety Standard Determination of vitamin A, D, E in food---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/GB5009.82-2016
GB
NATIONAL STANDARD
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
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.
3.1.2 Ascorbic acid (C6H8O6).
3.1.3 Potassium hydroxide (KOH).
3.1.4 Diethyl ether [(CH3CH2)2O]. it shall not contain peroxides after inspection,
AND the inspection method is as shown in A.2.
3.2 Reagent preparation
3.2.1 Potassium hydroxide solution (50 g/100 g). WEIGH 50 g of potassium
hydroxide; ADD 50 mL of water to dissolve it; after cooling it down, STORE it in
a polyethylene bottle.
3.2.2 Petroleum ether - ether solution (1 +1). MEASURE 200 mL of petroleum
ether; ADD 200 mL of ether; MIX it uniformly.
3.2.3 Organic filter head (pore size of 0.22 μm).
3.3 Standard substance
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).
4 Instruments and equipment
4.1 Analytical balance. the sensitivity is 0.01 mg.
4.2 Constant temperature water bath oscillator.
4.3 Rotary evaporator.
4.4 Nitrogen blowing instrument.
4.5 UV spectrophotometer.
4.7 High performance liquid chromatography. equipped with UV detector or
diode array detector or fluorescence detector.
5 Analytical procedures
5.1 Preparation of test specimens
A certain amount of samples are divided and crushed homogeneously, stored
in the sample bottle under refrigerated conditions in the dark; AND it shall be
used for determination as soon as possible.
5.2 Sample treatment
Caution. All vessels used shall not contain oxidizing substances. The
surface of the separation funnel piston glass shall not be coated with oil.
During the treatment, AVOID ultraviolet light and AVOID light as far as
possible. The extraction process shall be conducted in the fume hood.
5.2.1 Saponification
5.2.1.1 Starch-free samples
WEIGH 2 g ~ 5 g (accurate to 0.01 g) of homogenized solid sample OR 50 g
(accurate to 0.01 g) of liquid sample into a 150 mL flat-bottomed flask; as for
the solid test specimen, it is required to add about 20 mL of warm water; MIX it
uniformly; then ADD 1.0 g of ascorbic acid and 0.1 g of BHT; MIX it uniformly;
ADD 30 mL of absolute ethanol; ADD 10 mL ~ 20 mL potassium hydroxide
solution while shaking; after mixing it uniformly, PLACE it in the 80 °C constant
temperature water bath for shaking and saponification for 30 min; after
saponification, immediately USE cold water to cool it to room temperature.
5.2.2 Extraction
USE 30 mL of water to transfer the saponification solution into a 250 mL
separation funnel; ADD 50 mL of petroleum ether-ether mixture; SHAKE it for
extraction for 5 min; TRANSFER the lower solution into another 250 mL
separation funnel. ADD 50 mL of petroleum ether-ether mixture and CONDUCT
extraction again; COMBINE the ether layers.
5.2.3 Washing
USE about 100 mL of water to wash the ether layer; REPEAT washing for about
3 times, until washing the ether layer to neutral (it may use the pH test strips to
test the pH value of the lower solution); REMOVE the lower aqueous phase.
5.2.4 Concentration
The washed ether layer is filtered through an anhydrous sodium sulfate (about
3 g) into a 250 mL rotary evaporator or nitrogen concentrator. USE about 15 mL
of petroleum ether to rinse the separation funnel and anhydrous sodium sulfate
for 2 times; INCLUDE it in the evaporation bottle; and CONNECT it to the rotary
evaporator or gas concentration instrument; PLACE it in 40 °C water bath for
reduced pressure distillation or air concentration; when the ether solution in the
bottle is about 2 mL, TAKE the evaporation bottle off; immediately USE nitrogen
to blow it to near dry.
5.3 Chromatographic reference conditions
The chromatographic reference conditions are listed below.
5.4 Production of standard curve
This method uses external standard method for quantitative purposes.
Respectively INJECT the vitamin A and vitamin E standard series working
solution into the high performance liquid chromatograph. MEASURE the
corresponding peak area. USE the peak area as the ordinate and the
concentration of the standard determination solution as the abscissa, DRAW
the standard curve, and CALCULATE the linear regression equation.
5.5 Determination of samples
After the sample solution is analyzed by high performance liquid chromatograph,
the peak area is measured. USE the external standard method to calculate its
concentration based on the aforementioned standard curve.
6 Expression of analytical results
The content of vitamin A or vitamin E in the sample is calculated in accordance
with the equation (1).
7 Precision
The absolute difference between the two independent determinations obtained
under repeatability conditions shall not exceed 10% of the arithmetic mean.
8 Others
When the sampling amount is 5 g AND the constant volume is 10 mL, the UV
detection limit of the vitamin A is 10 μg/100 g AND the limit of quantification is
30 μg/100 g; the detection limit of the tocopherol is 40 μg/100 g AND the limit
of quantification is 120 μg/100 g.
9 Principles
The vitamin E in the sample was extracted with organic solvent, concentrated,
separated by high performance liquid phase chromatography amide column or
silica gel column, detected by fluorescence detector, AND quantified by external
standard method.
10 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.
10.1 Reagents
10.1.1 Absolute ethanol (C2H5OH). Chromatographic purity, it shall not contain
aldehydes after inspection, AND the inspection method is as shown in A.1.
10.1.2 Diethyl ether [(CH3CH2)2O]. Analytical grade pure, it shall not contain
oxides after inspection, AND the inspection method is as shown in A.1.
10.2 Reagent preparation
10.2.1 Petroleum ether - ether solution (1 +1). MEASURE 200 mL of petroleum
ether; ADD 200 mL of ether; MIX it uniformly; PREPARE it before use.
10.2.2 Mobile phase. n-hexane + [tert-butyl methyl ether - tetrahydrofuran -
methanol mixture (20 + 1 + 0.1)] = 90 +10; PREPARE it before use.
10.3 Standard substance
10.4 Preparation of standard solutions
10.4.1 Vitamin E standard stock solution (1.00 mg/mL). respectively and
accurately WEIGH 50.0 mg of each of α-tocopherol, β-tocopherol γ-tocopherol
and δ-Tocopherol (accurate to 0.1 mg);
11 Instruments and equipment
11.1 Analytical balance. the sensitivity is 0.1 mg.
11.2 Constant temperature water bath oscillator.
11.3 Rotary evaporator.
11.4 Nitrogen blowing instrument.
11.7 High performance liquid chromatograph with fluorescence detector or UV
detector.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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