GB 5009.250-2016 PDF English
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GB 5009.250-2016: National food safety standard ----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/GB5009.250-2016
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standard -
Determination of ethyl maltol in food
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 Principle ... 4
3 Reagents and materials ... 4
4 Instruments and equipment... 5
5 Analytical procedures ... 5
6 Expression of analytical results ... 7
7 Precision ... 8
8 Others ... 8
Appendix A High-performance liquid chromatogram of ethyl maltol ... 9
Appendix B Confirmation test... 10
National food safety standard -
Determination of ethyl maltol in food
1 Scope
This standard specifies the method for the determination of ethyl maltol in food
by high-performance liquid chromatography.
This standard applies to the determination of content of ethyl maltol in
beverages, candies, jellies, meat products, biscuits, bread, cakes, milk powder
foods.
2 Principle
After the specimen is extracted and purified, it is detected by a high-
performance liquid chromatograph which is equipped with a diode array
detector or a UV detector, quantified by an external standard method. The
positive sample needs to be qualitatively confirmed by mass spectrometry.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are of analytical
grade; the water is the grade II water as specified in GB/T 6682.
3.1 Reagents
3.1.1 Methanol (CH3OH). Chromatographically pure.
3.1.2 Acetonitrile (CH3CN). Chromatographically pure.
3.1.3 Sodium dihydrogen phosphate (NaH2PO4 • 2H2O).
3.2 Preparation of sodium dihydrogen phosphate solution
WEIGH 3.90 g of sodium dihydrogen phosphate; ADD water to dissolve and
dilute it to about 1000 mL; USE phosphoric acid to adjust the pH to 4.0 ± 0.1;
MAKE its volume reach to 1000 mL; USE a microporous membrane to filter it
to prepare for use.
3.3 Ethyl maltol (C7H8O3) standard substance
The purity is not less than 99%.
3.4 Preparation of standard solution
3.4.1 Standard stock solution. WEIGH 0.1 g of ethyl maltol (accurate to 0.0001
g); USE methanol to dissolve it and make its volume reach to the mark in a 100
mL volumetric flask. The concentration of this solution is 1 mg/mL.
3.4.2 Standard series working solution. Respectively and accurately TAKE
different volume of standard stock solution; USE methanol to dilute it to the
standard working solution which has a content of ethyl maltol of 0.0 μg/mL, 0.5
μg/mL, 2.0 μg/mL, 5.0 μg/mL, 25.0 μg/mL, 100.0 μg/mL.
4 Instruments and equipment
4.1 High-performance liquid chromatograph (HPLC). It is equipped with a diode
array detector (DAD) or UV detector (UVD).
4.2 Ultrasonic cleaner.
4.3 Vortex mixer.
4.4 Water bath.
4.5 Centrifuge. The speed is not less than 6000 r/min.
4.6 Analytical balance. Sensitivity is 0.01 g, 0.001 g, 0.0001 g, respectively.
5 Analytical procedures
5.1 Preparation of specimen
5.1.1 Specimens of carbonated drinks, fruit drinks, milk drinks, vegetable
protein drinks
Accurately WEIGH 10 g of specimen (accurate to 0.01 g) (carbonated beverage
needs to be ultrasonic for 2 min ~ 3 min to remove carbon dioxide before being
sampled) in a 25 mL stoppered test tube; USE acetonitrile to make its volume
reach to the mark; MIX it uniformly; ULTRASONIC it for 10 min (if sample
solution is turbid, CENTRIFUGE it at 6000 r/min for 10 min); TAKE the
supernatant; USE the microporous membrane to filter it; LOAD the filtrate into
the liquid chromatography for analysis.
5.1.2 Candies and jelly specimens
Accurately WEIGH 2 g of specimen (accurate to 0.001 g) in a 25 mL stoppered
test tube; ADD 20 mL of water; PUT it in a water bath at 60 °C ~ 70 °C to heat
5.2.4 Column’s temperature. 30 °C.
5.2.5 Detection wavelength. 276 nm.
5.2.6 Injection volume. 10 μL.
5.3 Production of standard curve
Respectively, INJECT the standard series working solution into the high-
performance liquid chromatograph, to determine the chromatographic peak
area of the corresponding ethyl maltol. USE the concentration of the standard
working solution as the abscissa and the peak area of the chromatographic
peak as the ordinate, to draw a standard curve.
5.4 Determination of sample solution
INJECT the sample solution into a high-performance liquid chromatograph, to
obtain the chromatographic peak area of the corresponding ethyl maltol.
According to the standard curve, OBTAIN the concentration of ethyl maltol in
the test solution.
The standard liquid chromatogram of ethyl maltol is as shown in Figure A.1.
5.5 Qualitative confirmation
When the specimen is determined, if the retention time of the chromatographic
peak of ethyl maltol is consistent with that of the standard substance,
meanwhile the ultraviolet absorption spectrum of this substance is consistent
with the ultraviolet absorption spectrum of the standard substance, it may
initially confirm the presence of the ethyl maltol to be determined in the
specimen. Positive specimen is subjected to a confirmation test by the use of a
mass spectrometer (see Appendix B).
6 Expression of analytical results
The content of ethyl maltol in the specimen is calculated according to formula
(1).
Where.
X - The content of ethyl maltol in the specimen, in milligrams per kilogram
(mg/kg);
c - The concentration of ethyl maltol in the sample solution which is obtained
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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