GB 5009.97-2023 English PDF (GB5009.97-2023)

Delivery: 3 seconds. Download true-PDF + Invoice.
Get QUOTATION in 1-minute: Click GB 5009.97-2023
Historical versions: GB 5009.97-2023
Preview True-PDF (Reload/Scroll if blank)

GB 5009.97-2023: National food safety standard - Determination of sodium cyclamate in foods
GB 5009.97-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards -- Determination of
cyclohexyl sulfamate in food
ISSUED ON. SEPTEMBER 06, 2023
IMPLEMENTED ON. MARCH 06, 2024
Issued by. National Health Commission of the People's Republic of China;
State Administration for Market Regulation.
Table of Contents
Foreword... 3
1 Scope... 4
2 Principle... 4
3 Reagents and materials... 4
4 Instruments and equipment... 5
5 Analysis steps... 6
6 Expression of analysis results... 8
7 Precision... 9
8 Other... 9
9 Principle... 9
10 Reagents and materials... 10
11 Instruments and equipment... 11
12 Analysis steps... 11
13 Expression of analysis results... 13
14 Precision... 13
15 Other... 13
16 Principle... 14
17 Reagents and materials... 14
18 Instruments and equipment... 15
19 Analysis steps... 16
20 Expression of analysis results... 19
21 Precision... 20
22 Other... 20
Annex A Gas chromatogram of cyclohexylaminosulfonic acid derivatives... 21
Annex B Liquid chromatogram of cyclohexylsulfamic acid derivatives... 22
Annex C Cyclohexylsulfamic acid multiple reaction monitoring (MRM) chromatogram
... 23
National food safety standards -- Determination of
cyclohexyl sulfamate in food
1 Scope
This Standard specifies the method for the determination of cyclohexyl sulfamate in
food.
Method One -- Gas Chromatography is applicable to the determination of cyclohexyl
sulfamate in food (except distilled wine, fermented wine, prepared wine, cooking wine
and other ethanol-containing foods).
Method Two -- Liquid Chromatography is applicable to the determination of cyclamate
in food.
Method Three -- Liquid Chromatography-Mass Spectrometry/Mass Spectrometry is
applicable to the determination of cyclamate in food.
Method One -- Gas Chromatography
2 Principle
The cyclamate in the specimen is extracted with water. It reacts with sodium nitrite in
sulfuric acid medium to generate cyclohexanol nitrite and cyclohexanol. After
extraction with n-heptane, use gas chromatography-hydrogen flame ionization detector
to determine. Use external standard method to quantify.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure, and the
water is grade one water specified in GB/T 6682.
3.1 Reagents
3.1.1 n-Heptane [CH3(CH2)5CH3]. chromatographically pure.
3.1.2 Petroleum ether. Boiling range is 30℃~60℃.
3.1.3 Sulfuric acid (H2SO4). 95.0%~98.0% (mass fraction); density. 1.84 g/mL.
3.1.4 Sodium nitrite (NaNO2).
3.1.5 Amylase. enzyme activity ≥1500 U/g.
3.1.6 Absolute ethanol (CH3CH2OH).
3.2 Preparation of reagents
3.2.1 Sodium nitrite solution (50 g/L). Weigh 50 g of sodium nitrite, dissolve in water
and dilute to 1000 mL. Mix well.
3.2.2 Sulfuric acid solution (200 g/L). Measure 108 mL of sulfuric acid and slowly add
it to 800 mL of water. Stir constantly to avoid localized overheating. After cooling, add
water to dilute to 1000 mL. Mix well.
3.3 Standard product
Sodium cyclohexyl sulfamate standard (C6H12NSO3Na). purity ≥99%, or a standard
certified by the country and awarded a reference material certificate.
3.4 Preparation of standard solution
3.4.1 Cyclohexylsulfamic acid standard stock solution (6 mg/mL). Accurately weigh an
appropriate amount of sodium cyclamate standard (accurate to 0.1 mg) into a 100 mL
volumetric flask. Dissolve in water and bring the volume to the mark. Mix well (the
conversion factor of sodium cyclohexylsulfamate into cyclohexylsulfamate is 0.8907).
Transfer the solution to a brown glass container. Store at 4°C away from light. It is valid
for 12 months.
3.4.2 Cyclohexylsulfamic acid standard intermediate solution (1200 μg/mL).
Accurately pipette 10 mL of cyclamate standard stock solution into a 50 mL volumetric
flask. Use water to bring the volume to the mark. Mix well. Transfer the solution to a
brown glass container. Store at 4°C away from light. It is valid for 6 months.
3.5 Materials
3.5.1 Centrifuge tube. 50 mL graduated centrifuge tube with screw cap.
3.5.2 Organic phase microporous filter membrane. 0.45 μm in pore size.
4 Instruments and equipment
4.1 Gas chromatograph. equipped with hydrogen flame ionization detector (FID).
4.2 Analytical balance. division is 1 mg, 0.1 mg.
4.3 Vortex mixer.
Vortex for 5 min. Perform ultrasonic extraction for 30 min. Mix well and bring to room
temperature.
5.2.2 Specimens of jelly, candy, rice flour, starch products, etc.
Weigh 2 g of specimen (accurate to 0.001 g) into a centrifuge tube. Add 20 mL of water
(0.2 g of amylase is required for specimens such as rice flour and starch products). After
mixing evenly, heat in a water bath at 60℃±2℃ for 20 min. Vortex for 5 min. Perform
ultrasonic extraction for 10 min. Mix well and bring to room temperature.
5.2.3 Other solid and semi-solid specimens
Weigh 2 g of specimen (accurate to 0.001 g) into a centrifuge tube. Add 20 mL of water.
Vortex for 5 min. Perform ultrasonic extraction for 30 min. Mix well and bring to room
temperature.
5.2.4 Liquid specimen
Weigh 2 g of specimen (accurate to 0.001 g) into a centrifuge tube. Add water to 20 mL.
Vortex for 5 min. Perform ultrasonic extraction for 10 min. Mix well and bring to room
temperature.
5.3 Derivatization
Place the centrifuge tube containing the specimen extract solution in an ice bath for 10
min, then add 10 mL of n-heptane, 5 mL of sodium nitrite solution (50 g/L), and 5 mL
of sulfuric acid solution (200 g/L) in sequence. Mix well. Place in ice bath for 30 min.
Shake 3 to 5 times during this period. Remove and vortex for 3 min. Centrifuge at 9000
r/min for 3 min at 4°C (if emulsification occurs, slowly add absolute ethanol dropwise.
At the same time, shake the centrifuge tube gently until the emulsification breaks.
Centrifuge at 9000 r/min for 3 mins at 4°C). Pass the supernatant through the organic
phase microporous filter membrane for measurement on the machine.
5.4 Preparation and derivatization of standard series working solutions
Accurately pipette 0.05 mL, 0.10 mL, 0.25 mL, 0.50 mL, 1.00 mL, 2.5 mL, and 5.0 mL
of cyclohexylaminosulfamic acid standard intermediate solution (1200 μg/mL) into
centrifuge tubes. Dilute to 20 mL with water and derivatize according to the
determination step in 5.3.At this time, the concentrations of the analytes in n-heptane
are respectively equivalent to 6 μg/mL, 12 μg/mL, 30 μg/mL, 60 μg/mL, 120 μg/mL,
300 μg/mL, and 600 μg/mL. Prepare when needed.
5.5 Instrument reference conditions
5.5.1 Chromatographic column. medium polar capillary column internally coated with
50% phenyl-50% methylpolysiloxane, 30 m × 0.32 mm × 0.25 μm or a column with
equivalent performance.
5.5.2 Column temperature rising program. initial temperature is 50°C and maintain for
3 min. Raise temperature to 70°C at 10°C/min and hold for 0.5 min. Raise the
temperature to 220°C at 30°C/min and hold for 3 min.
5.5.3 Inlet temperature. 230℃.
5.5.4 Injection volume. 1 μL.
5.5.5 Injection method. split injection, with a split ratio of 1.5.
5.5.6 Detector. hydrogen flame ionization detector (FID), with a temperature of 260℃.
5.5.7 Carrier gas. high-purity nitrogen, with a flow rate of 2.0 mL/min.
5.5.8 Hydrogen. 32 mL/min. Air. 300 mL/min.
5.6 Preparation of standard curve
The derivatized standard series working solution is injected into the gas chromatograph.
Th...