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GB 5009.249-2016 PDF in English


GB 5009.249-2016 (GB5009.249-2016) PDF English
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GB 5009.249-2016: PDF in English

GB 5009.249-2016
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standard -
Determination of ferrous sodium
ethylenediaminetetraacetate in iron fortified soy sauce
ISSUED ON. AUGUST 31, 2016
IMPLEMENTED ON. MARCH 01, 2017
Issued by. National Health and Family Planning Commission of the PRC
Table of Contents
Foreword ... 3 
1 Scope ... 4 
2 Principles ... 4 
3 Reagents and materials ... 4 
4 Instruments and equipment... 5 
5 Analytical procedures ... 5 
6 Expression of analytical results ... 6 
7 Precision ... 7 
8 Others ... 7 
Appendix A Liquid chromatogram of ferrous sodium
ethylenediaminetetraacetate standard solution ... 8 
National food safety standard -
Determination of ferrous sodium
ethylenediaminetetraacetate in iron fortified soy sauce
1 Scope
This standard specifies the method for the determination of ferrous sodium
ethylenediaminetetraacetate in iron fortified soy sauce.
This standard applies to the determination of ferrous sodium
ethylenediaminetetraacetate in iron fortified soy sauce.
2 Principles
The specimen is precipitated by methanol and filtered, the filtrate is diluted and
separated by reversed-phase high-performance liquid chromatography. It is
detected by the ultraviolet detector at a wavelength of 254 nm, the retention
time of the chromatographic peak is used for qualitative determination, the
external standard method is used for quantification.
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
3.1.1 Methanol (CH3OH).
3.1.2 Methanol (CH3OH). chromatographically pure.
3.1.3 Water solution of tetrabutylammonium hydroxide (C16H36N • OH) (40
g/100 mL).
3.1.4 Formic acid (HCOOH).
3.1.5 Formic acid solution (100 mL/L). PIPETTE 5 mL of formic acid; USE water
to make its volume reach to 50 mL.
3.1.6 Aqueous methanol solution (3 + 1). TAKE 750 mL of methanol (3.1.1);
ADD 250 mL of water; MIX it uniformly.
3.1.7 Ferrous sodium ethylenediaminetetraacetate standard substance
(C10H12FeN2NaO8 • 3H2O). purity ≥ 99.0%.
3.2 Preparation of standard solution
3.2.1 Ferrous sodium ethylenediaminetetraacetate standard stock solution (2.0
mg/mL). WEIGH 0.05 g of ferrous sodium ethylenediaminetetraacetate
standard substance (accurate to 0.0001 g); PLACE it in a 25 mL volumetric
flask; USE water to dissolve it to make it reach to a certain volume. This solution
shall be prepared in a brown volumetric flask, stored in a refrigerator in the dark,
recommended for use within 15 days.
3.2.2 Ferrous sodium ethylenediaminetetraacetate standard intermediate
solution (100 μg/mL). USE the aqueous methanol solution (3.1.6) to make the
volume of 2.5 mL of ferrous sodium ethylenediaminetetraacetate standard
stock solution to 50 mL. This solution shall be prepared in a brown volumetric
flask and stored in the refrigerator in the dark.
3.2.3 Ferrous sodium ethylenediaminetetraacetate standard working solution.
Respectively, PIPETTE 1.0 mL, 2.0 mL, 4.0 mL, 6.0 mL, 8.0 mL, 10.0 mL of
ferrous sodium ethylenediaminetetraacetate standard intermediate solution in
a 50.0 mL brown volumetric flask; USE water to make its volume reach to the
mark. The concentration is 2.0 μg/mL, 4.0 μg/mL, 8.0 μg/mL, 12.0 μg/mL, 16.0
μg/mL, 20.0 μg/mL, which shall be prepared before use.
4 Instruments and equipment
4.1 High-performance liquid chromatograph, which is equipped with UV
detector or diode array detector.
4.2 Balance. The sensitivity is 0.1 mg.
4.3 pH meter. Accuracy is 0.01.
5 Analytical procedures
5.1 Preparation of sample solution
PIPETTE 2.50 mL of the uniformly-mixed specimen; PLACE it into a 50.0 mL
brown volumetric flask; ADD aqueous methanol solution (3.1.6) to dilute it to
the mark; SHAKE it uniformly. LET it be standing in the dark for 50 min; USE
filter paper to filter it. PIPETTE 5.00 mL of filtrate; PLACE it into a 50 mL brown
volumetric flask; USE water to dilute it to the mark; SHAKE it uniformly; USE
0.45 μm filter membrane to filter it, to obtain a sample solution, which is ready
......
Source: Above contents are excerpted from the PDF -- translated/reviewed by: www.chinesestandard.net / Wayne Zheng et al.