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GB 5009.247-2025: National food safety standard - Determination of Neotame in foods
Status: Valid GB 5009.247: Evolution and historical versions
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National food safety standard - Determination of Neotame in foods
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GB 5009.247-2025
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| GB 5009.247-2016 | English | 80 |
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Method for the determination of Neotame in foods -- High-performance liquid chromatography
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GB 5009.247-2016
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Basic data | Standard ID | GB 5009.247-2025 (GB5009.247-2025) | | Description (Translated English) | National food safety standard - Determination of Neotame in foods | | Sector / Industry | National Standard | | Classification of Chinese Standard | X09 | | Word Count Estimation | 13,191 | | Issuing agency(ies) | National Health Commission of the People's Republic of China, State Administration for Market Regulation | GB 5009.247-2016: Method for the determination of Neotame in foods -- High-performance liquid chromatography
---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
(Food safety national standard - Determination of neotame in foods)
National Standards of People's Republic of China
National Food Safety Standard
DETERMINATION neotame
Issued on. 2016-08-31
2017-03-01 implementation
People's Republic of China
National Health and Family Planning Commission released
Foreword
This standard replaces GB/T 23378-2009 "Determination of foods neotame by HPLC."
This standard compared with GB/T 23378-2009, the main changes are as follows.
--- Standard name was changed to "national food safety standards in food Neotame determination."
--- Scope Change drinks, sweets, cakes, roasted seeds and nuts, pickles, candy, jams, jellies, sauces complex measured neotame.
National Food Safety Standard
DETERMINATION neotame
1 Scope
This standard specifies a high performance liquid chromatography in food neotame content.
This standard applies to the determination of drinks, sweets, cakes, roasted seeds and nuts, pickles, candy, jams, jellies, seasonings in food neotame.
Principle 2
The sample was mixed extraction liquid extraction, solid phase extraction (SPE), high performance liquid chromatograph retention time qualitative, external standard peak area
Statutory amount.
3 Reagents and materials
Unless otherwise indicated, the methods used were of analytical grade reagents and water as a water GB/T 6682 regulations.
3.1 Reagents
3.1.1 acetonitrile (CH3CN). chromatography.
3.1.2 octane sulfonate (C8H17NaO3S). chromatography.
3.1.3 phosphoric acid (H3PO4).
3.1.4 formic acid (CH3COOH). chromatography.
3.1.5 Methanol (CH3OH). chromatography.
3.1.6 triethylamine (C6H15N). chromatography.
3.2 reagent preparation
3.2.1 Mixed extracts. Pipette 0.8mL and 2.5mL of triethylamine formate, add water to 1000mL, pH of about 4.5.
3.2.2 Ion pair reagent buffer. Weigh 2.00g octane sulfonate, was dissolved with 500mL of water, phosphoric acid was added 1.0mL, add water to
1000mL.
3.3 Standard
Neotame (C20H30N2O5, CAS number. 165450-17-9), purity ≥99.0%.
3.4 Standard Solution
3.4.1 Standard stock solution. Accurately weigh 0.1000g neotame standards, plus mixed extract dissolved volume to 100mL, the solution containing neotame
An amount of 1.00mg/mL.
3.4.2 Working standard solution. Pipette appropriate amount of neotame standard stock solution, mixed with extracts formulated to concentrations of 0.2μg/mL,
1.0μg/mL, 5.0μg/mL, 10.0μg/mL, 50.0μg/mL, 100.0μg/mL series of standard working solution.
3.5 C18 solid phase extraction column. 6mL, 500mg, or rather who, prior to use successively with methanol 5mL, 10mL water activation.
3.6 0.45μm membrane, organic.
4 instruments and equipment
4.1 liquid chromatograph equipped with an ultraviolet detector or diode array detector.
4.2 ultrasonic cleaning.
4.3 Analysis of balance. a sense of volume and 0.0001g 0.01g.
4.4 Organization mashed machines.
4.5 Vortex.
4.6 nitrogen blowing instrument.
4.7 Solid phase extraction device.
4.8 centrifuge. speed ≥4000r/min.
Step 5 Analysis
5.1 Sample Preparation
5.1.1 solid samples
Weigh 10g (accurate to 0.01g) in 50mL sample pulverized uniformly stoppered plastic centrifuge tube, add 30mL mixed extract, spin
Vortex oscillation 10min, ultrasound 30min, with a mixed extract to the mark, if the solution is cloudy, not less than 4000r/min centrifugal
10min after filtration and set aside.
5.1.2 Liquid Sample
Accurately weighed sample in 50mL 10.0mL stoppered plastic centrifuge tube, add 30mL mixed extract Shakers. Ultrasonic
After 15min, the mixed extract to the mark, if the solution is cloudy, not less than 4000r/min centrifugal 10min, filtered and set aside.
Note. Samples containing gas such as carbonated beverages, soft drinks and other first lukewarm, stirring to remove the carbon dioxide in the sample or ultrasonic degassing, and then accurately weighed sample.
5.2 Sample purification
Absorb 10.0mL filtrate flow rate of 1mL/min ~ 2mL/min through a solid phase extraction column until the filtrate flows out completely by
The extract was mixed with a flow rate of 5mL 1mL/min ~ 2mL/min leaching extraction column, discard all effluent with 5mL methanol
1mL/min flow rate were eluted at 40 ℃ water bath with nitrogen blowing instrument concentrated extract volume to 2.0mL with a mixed, through 0.45μm
After membrane filtration as a test solution for liquid chromatography analysis.
5.3 Instrument Reference conditions
5.3.1 Column. C18 column, 5μm, 250mm × 4.6mm (diameter) or equivalent person.
5.3.2 Column temperature. 30 ℃.
5.3.3 Mobile phase. A phase. acetonitrile; B phase. ion pair reagent buffer. Gradient program shown in Table 1.
5.3.4 flow rate. 1.0mL/min.
5.3.5 Detection wavelength. 218nm.
5.3.6 Injection volume. 50μL.
Table 1 gradient elution program
Time/min Mobile phase A /% mobile phase B /%
0.00 25.0 75.0
10.00 65.0 35.0
20.00 100.0 0.0
24.00 100.0 0.0
24.10 25.0 75.0
30.00 25.0 75.0
5.4 standard curve
The standard series working solution were injected into the liquid chromatograph to measure the peak area corresponding to the concentration of the working standard solution as abscissa and the
Peak area of the vertical axis, the standard curve (Neotame standard chromatogram shown in Figure A.1).
5.5 Determination of the sample solution
The sample solution into the liquid chromatograph retention time qualitative, and record the peak area to obtain test solution based on the standard curve Zealand
Sweet concentration response in the sample solution neotame content should be within the linear range of the instrument.
6 expression analysis
Neotame content of the sample according to the formula (1).
X =
c × V × V1 × 1000
m × V2 × 1000
(1)
Where.
X --- sample neotame content in milligrams per kilogram (mg/kg) or milligrams per liter (mg/L);
C --- concentration of the sample solution neotame, in micrograms per milliliter (μg/mL);
The V --- eluate was concentrated sample volume to volume, in milliliters (mL of);
Vl --- volume of the sample extract volume in milliliters (mL of);
m --- sample weight in grams (g);
--- Draw volume V2 of the filtrate in milliliters (mL).
The results to two significant figures.
7 precision
Two independent determination results under the absolute difference in repeatability condition must not exceed 10% of the arithmetic mean.
8 Other
The limit of quantification of the standard methods of 0.2mg/kg.
Appendix A
Chromatogram
Neotame standard chromatogram shown in Figure A.1.
Figure A.1 Neotame standard liquid chromatogram
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