GB 5009.97-2016 English PDF (GB5009.97-2016)

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GB 5009.97-2016: Determination of sodium cyclamate in foods
GB 5009.97-2016
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard -
Determination of Sodium Cyclamate in Foods
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 ... 4 
1 Scope ... 5 
2 Principle ... 5 
3 Reagents and materials ... 6 
4 Instruments and equipment ... 7 
5 Analytical procedures ... 7 
6 Expression of analysis results ... 10 
7 Precision ... 10 
8 Other ... 10 
9 Principle ... 11 
10 Reagents and materials ... 11 
11 Instruments and equipment ... 12 
12 Analytical procedures ... 12 
13 Expression of analysis results ... 14 
14 Precision ... 15 
15 Other ... 15 
16 Principle ... 15 
17 Reagents and materials ... 15 
18 Instruments and equipment ... 16 
19 Analytical procedures ... 16 
20 Expression of analysis results ... 18 
21 Precision ... 18 
22 Other ... 19 
Appendix A Reference conditions of liquid chromatography-mass
spectrometry/mass spectrometry ... 20 
Appendix B Chromatograms of sodium cyclamate standard solution ... 22 
National Food Safety Standard -
Determination of Sodium Cyclamate in Foods
1 Scope
This Standard specifies three methods for the determination of sodium
cyclamate in foods - gas chromatography, liquid chromatography, and liquid
chromatography-mass spectrometry/mass spectrometry. [Translator note:
There is additional 1 nick-name of sodium cyclamate in Chinese]
Gas chromatography of this Standard applies to the determination of sodium
cyclamate in beverages, preserved fruits, fruit leather, pickled fruits, shelled and
unshelled cooked nuts and seeds, canned fruits, jams, pastries, bread, biscuits,
frozen drinks, jelly, compound seasonings, pickled vegetables, and fermented
bean curd foods.
Gas chromatography of this Standard does not apply to the determination of
this compound in white spirits.
Liquid chromatography of this Standard applies to the determination of sodium
cyclamate in beverages, preserved fruits, fruit leather, pickled fruits, shelled and
unshelled cooked nuts and seeds, compound wine, canned fruits, jams,
pastries, bread, biscuits, frozen drinks, jelly, compound seasonings, pickled
vegetables, and fermented bean curd foods.
Liquid chromatography-mass spectrometry/mass spectrometry of this Standard
applies to the determination of sodium cyclamate in white spirit, wine, yellow
rice wine, and cooking wine.
Method 1 -- Gas chromatography
2 Principle
The sodium cyclamate in the food is extracted using water. In a sulfuric acid
medium, sodium cyclamate is reacted with nitrous acid, to form cyclohexanol
nitrite. USE gas chromatograph-hydrogen flame ionization detector for
separation and analysis. USE the retention time for qualitation, external
standard method to quantify.
Accurately WEIGH 0.5612 g of sodium cyclamate standard; USE water to
dissolve and dilute to 100 mL; MIX well. 1.00 mL of this solution is equivalent
to 5.00 mg of cyclohexylsulfamic acid (The conversion factor of sodium
cyclamate and cyclohexylsulfamic acid is 0.8909). STORE in a refrigerator at
1 °C~4 °C. It can be stored for 12 months.
3.4.2 Standard use solution of cyclohexylsulfamic acid (1.00 mg/mL):
Accurately PIPETTE 20.0 mL of standard stock solution of cyclohexylsulfamic
acid; USE water to dilute to 100 mL; MIX well. STORE in a refrigerator at
1 °C~4 °C. It can be stored for 6 months.
4 Instruments and equipment
4.1 Gas chromatograph: Equipped with a hydrogen flame ionization detector
(FID).
4.2 Vortex mixer.
4.3 Centrifuge: Speed≥4000 r/min.
4.4 Ultrasonic oscillator.
4.5 Sample pulverizer.
4.6 10 μL micro syringe.
4.7 Thermostat water bath.
4.8 Balance: The sensitivity is 1 mg, 0.1 mg.
5 Analytical procedures
5.1 Preparation of sample solutions
5.1.1 Liquid sample processing
5.1.1.1 After shaking the ordinary liquid sample well, WEIGH 25.0 g of the
sample (may be filtered if necessary); USE water to dilute to 50 mL for use.
5.1.1.2 Carbon dioxide-containing sample: WEIGH 25.0 g of the sample in a
beaker; HEAT it in a 60 °C water bath for 30 min to remove carbon dioxide; LET
cool; USE water to dilute to 50 mL for use.
5.1.1.3 Alcohol-containing sample: WEIGH 25.0 g of the sample in a beaker;
USE sodium hydroxide solution (3.2.1) to adjust to a weak alkaline pH of 7~8;
low temperature for 20 min to clear stratification, the supernatant is taken and
stored in a refrigerator at 1 °C~4 °C for sample injection.
5.2 Preparation and derivatization of standard solution series
Accurately PIPETTE 0.50 mL, 1.00 mL, 2.50 mL, 5.00 mL, 10.0 mL, 25.0 mL of
1.00 mg/mL standard solution of cyclohexylsulfamic acid in 50 mL volumetric
flasks; ADD water to dilute to the mark. Prepare the standard solution series,
with concentrations of 0.01 mg/mL, 0.02 mg/mL, 0.05 mg/mL, 0.10 mg/mL, 0.20
mg/mL, and 0.50 mg/mL. Prepare when used for derivatization.
Accurately PIPETTE 10.0 mL of the standard series solution. The derivatization
is same as (5.1.3).
5.3 Determination
5.3.1 Chromatographic conditions
5.3.1.1 Chromatographic column: Weakly polar quartz capillary column
(internally coated with 5% phenyl methyl polysiloxane, 30 m×0.53 mm×1.0 μm)
or an equivalent column.
5.3.1.2 Column temperature heating program: Initial temperature 55 °C is kept
for 3 min. At 10 °C/min, the temperature is raised to 90 °C and kept for 0.5 min.
At 20 °C/min, the temperature is raised to 200 °C and kept for 3 min.
5.3.1.3 Injection port: Temperature is 230 °C. Injection volume is 1 μL,
splitless/split injection. Split ratio is 1:5 (Split ratio and mode may be adjusted
according to chromatographic instrument conditions).
5.3.1.4 Detector: Hydrogen flame ionization detector (FID). Temperature is
260 °C.
5.3.1.5 Carrier gas: High-purity nitrogen. Flow rate is 12.0 mL/min. Make-up
flow rate is 20 mL/min.
5.3.1.6 Hydrogen: 30 mL/min; air 330 mL/min (The flow rate of the carrier gas,
hydrogen, air may be adjusted according to the instrument conditions).
5.3.2 Chromatographic analysis
PIPETTE 1 μL of the supernatant of derivatization-treated standard series
solutions of each concentration (5.2); inject them into the gas chromatograph.
The peak area of the response value of different concentrations of the analyte
can be measured. USE the concentration as the abscissa. USE the sum of the
areas of the two peaks of cyclohexanol nitrite and cyclohexanol as the ordinate.
DRAW the standard curve.
Method 2 -- High-performance liquid chromatography
9 Principle
After the sodium cyclamate in the food is extracted using water, in a strong
acidic solution, it is reacted with sodium hypochlorite, to form N,N-
dichlorocyclohexylamine. After using n-heptane to extract, USE high-
performance liquid chromatography to detect; USE the retention time for
qualitation, external standard method to quantify.
10 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure;
the water is the Grade 1 water specified in GB/T 6682.
10.1 Reagents
10.1.1 N-heptane [CH3(CH2)5CH3]: Chromatographically pure.
10.1.2 Acetonitrile (CH3CN): Chromatographically pure.
10.1.3 Sulfuric acid (H2SO4).
10.1.4 Sodium hypochlorite (NaClO).
10.1.5 Sodium bicarbonate (NaHCO3).
10.1.6 Zinc sulfate (ZnSO4 • 7H2O).
10.1.7 Potassium ferrocyanide {K4[...