GB 5009.140-2023National food safety standard - Determination of acesulfame K in food (English PDF)

3.1 Reagents

3.2 Preparation of reagents

3.2.1 Potassium ferrocyanide solution (92 g/L). Weigh 106 g of potassium ferrocyanide,

add water to dissolve, and dilute to 1000 mL.

3.2.2 Zinc acetate solution (183 g/L). Weigh 219 g of zinc acetate, dissolve in a small

amount of water, add 32 mL of glacial acetic acid, and dilute to 1000 mL with water.

3.3 Reference material

Acesulfame potassium reference material (C4H4KNO4S, CAS number. 55589-62-3),

purity > 99.0 %, or a reference material certified by the country and awarded a reference

material certificate.

3.4 Preparation of standard solutions

3.4.1 Acesulfame potassium standard stock solution (1000 mg/L). Accurately weigh

3.5 Materials

3.5.1 Neutral alumina solid-phase extraction column (6 mL, 500 mg), activated with 10

mL of acetonitrile before use, and then balanced with 10 mL of water.

3.5.2 Water phase microporous filter membrane. 0.45 µm.

3.5.3 Plastic centrifuge tube. 50 mL.

4.1 High-performance liquid chromatograph. equipped with a UV detector or a diode

array detector.

4.6 Constant temperature water bath.

4.7 Ultrasonic generator.

4.8 Solid-phase extraction device.

4.9 Homogenizer.

5.1 Preparation of samples

Homogeneous liquid samples are mixed directly, and gas-containing samples are

transferred to a beaker before use and degassed in a 50 °C water bath for 10 minutes or

ultrasonic for 5 minutes; non-homogeneous liquid and semi-solid samples are

homogenized with a homogenizer; solid samples are ground thoroughly with a food

grinder and mix well.

5.2 Treatment of samples

5.2.1 Beverages (except protein drinks), fruit products, vegetable products

Weigh about 5 g (accurate to 0.001 g) of the sample in a 50 mL centrifuge tube, add 20

mL of water, vortex to mix well, sonicate for 20 minutes, centrifuge at 10000 r/min for

5.2.2 Other samples

Weigh 5 g (accurate to 0.001 g) of the sample in a 50 mL centrifuge tube, add 20 mL of

water, vortex to mix well, and sonicate for 20 minutes.

5.3 Instrument reference conditions

5.3.1 Chromatographic column. C18 column (column length 250 mm, inner diameter

4.6 mm, particle size 5 µm), or a chromatographic column with equivalent performance.

5.3.3 Flow rate. 1.0 mL/min.

5.3.4 Column temperature. 30 degrees C.

5.4 Plotting of standard curve

Time Phase A Phase B

Inject the standard series working solutions into the high-performance liquid

chromatograph in order from low to high concentration, measure the corresponding

peak area, and plot the standard curve with the concentration of acesulfame potassium

in the standard working solutions as the abscissa and the peak area as the ordinate. For

the liquid chromatogram of acesulfame potassium standard solutions, see Figure A.1 in

Annex A.

5.5 Determination of sample solution

Inject the sample solution into the high-performance liquid chromatograph, make

qualitative identification based on the retention time, measure the peak area, and obtain

the concentration of acesulfame potassium in the sample solution according to the

standard curve.