Issued on: September 6, 2023 Implemented on: March 6, 2024

Issued by: National Health Commission of the People's Republic of China;

State Administration for Market Regulation.
Issued on: September 6, 2023

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 National Food Safety Standard - Determination of Sucralose (Sucralose) in Food

Foreword
This standard replaces GB 22255-2014 National Food Safety Standard - Determination of
Sucralose (Sucralose) in Food. Compared with GB 22255-2014, the main changes of this
standard are as follows:
• Added the second method "High Performance Liquid Chromatography-Tandem Mass
Spectrometry";
• Modified the mobile phase conditions for the high performance liquid chromatography
with refractive index detector;
• Modified the sample pretreatment conditions for high performance liquid chromatography;

• Modified the detection limit and quantification limit for high performance liquid
chromatography.

1 Scope
This Standard specifies the high performance liquid chromatography and high
performanceliquid chromatography - tandem mass spectrometry for the determination of
sucralose in foods.
Method I -High Performance Liquid Chromatography

2 Principle
The sucralose in the specimen is extracted with methanol-water solution to remove protein
andfat. After purification by a solid phase extraction column, evaporation to dryness,
redissolutionand enrichment, a high-performance liquid chromatograph is used, and a
reversed-phase Cischromatographic column is used for separation. A differential detector or
an evaporative light.scattering detector is adopted for detection. In accordance with the
retention time of thechromatographic peak, conduct qualitative determination. Adopt the
external standard methodfor quantitative determination.

3 Reagents and Materials

Unless otherwise specified, all reagents used in this method are of analytical purity, and
water conforms to grade I water specified in GB/T 6682.
3.1 Reagents
3.1.1 Methanol (CH₃OH).
3.1.2 Methanol (CH₃OH): chromatographically pure.

3.1.3 n-Hexane (C₆ H₁₄).

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3.1.4 Dipotassium hydrogen phosphate (K₂HPO₄·3H₂O).
3.1.5 Zinc acetate [Zn(CH₃COO)₂·2H₂O].
3.1.6 Potassium ferrocyanide [K₄Fe(CN)₆ ·3H₂O].
3.1.7 Acetic acid (CH₃COOH).

3.1.8 Acetonitrile (CH₃CN): chromatographically pure.

3.1.9 Neutral aluminum oxide: particle size 75μm–150μm (100 mesh–200 mesh).
3.2 Reagent Preparation
3.2.1 Zinc acetate solution (219 g/L): Weigh 21.9 g of zinc acetate, add 3 mL of acetic acid,
and dissolve in water to make 100 mL.
3.2.2 Potassium ferrocyanide solution (106 g/L): Weigh 10.6 g of potassium ferrocyanide,
and dissolve in water to make 100 mL.
3.2.3 Dipotassium hydrogen phosphate solution (0.1%): Weigh 0.1 g of dipotassium
hydrogen phosphate, and dissolve in water to make 100 mL.
3.2.4 Methanol-0.1% dipotassium hydrogen phosphate solution (20+80): Mix methanol and
0.1% dipotassium hydrogen phosphate solution in a volume ratio of 20:80.
3.2.5 Methanol-water solution (75+25): Mix methanol and water in a volume ratio of 75:25.
3.2.6 Water-acetonitrile solution (89+11): Mix water and acetonitrile in a volume ratio of
89:11.
3.3 Reference Material
Sucralose standard substance (C₁₂H₁₉ Cl₃O₈ , CAS No.: 56038-13-2) with a purity of ≥99%,
or a standard substance certified by the state and issued with a standard substance
certificate.
3.4 Preparation of Standard Solutions
3.4.1 Sucralose standard stock solution (10.0 mg/mL): Weigh 0.25 g of sucralose standard
substance (accurate to 0.0001 g), dissolve in water, transfer to a 25 mL volumetric flask,
dilute to the mark, and mix well. Store the stock solution in a refrigerator at 4°C with a shelf
life of 6 months.
3.4.2 Sucralose standard intermediate solution (1.00 mg/mL): Pipette 5.00 mL of the
sucralose standard stock solution (10.0 mg/mL) into a 50 mL volumetric flask, dilute to the
mark with water, and mix well. Store in a refrigerator at 4°C with a shelf life of 3 months.
3.4.3 Sucralose standard series working solutions: Pipette an appropriate amount of the
sucralose standard intermediate solution (1.00 mg/mL) and dilute with water to prepare
standard series working solutions with mass concentrations of 0.0200 mg/mL, 0.0500 mg/mL,
0.100 mg/mL, 0.200 mg/mL, 0.400 mg/mL, 0.800 mg/mL, and 1.00 mg/mL. Prepare fresh
before use.
3.5 Materials

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3.5.1 Solid-phase extraction column (specification: 200 mg/6 mL, packed with N-
vinylpyrrolidone and divinylbenzene hydrophilic-lipophilic balanced filler or equivalent).
3.5.2 0.45 μm hydrophilic microporous membrane and 0.45 μm hydrophobic microporous
membrane.

4 Instruments and Equipment

4.1 High performance liquid chromatograph: equipped with a refractive index detector or an
evaporative light scattering detector.
4.2 Balance: with sensitivities of 0.1 mg and 1 mg respectively.
4.3 Vortex mixer.
4.4 Water bath.
4.5 Ultrasonic generator: 50 kHz.
4.6 Centrifuge: rotational speed ≥3000 r/min.
4.7 High-speed centrifuge: rotational speed ≥10000 r/min.
4.8 Solid-phase extraction device.
4.9 Homogenizer.
4.10 Crusher.

5 Analytical Procedures
5.1 Sample Preparation
Liquid samples are shaken well; uniformly matrixed semi-solid and powdered samples are
directly determined; other samples need to be homogenized or crushed to uniformity.
5.2 Sample Extraction
5.2.1 Alcoholic Samples
5.2.1.1 Liquid alcoholic samples
Weigh 2 g–5 g of the mixed alcoholic sample (accurate to 0.001 g) into an evaporating dish,
evaporate to dryness in a boiling water bath, dissolve the residue with 1.00 mL of water, filter
through a 0.45 μm hydrophilic microporous membrane, and the filtrate is the prepared
sample solution for testing.
5.2.1.2 Alcohol specimens containing solid substance
Weigh-take 2 g~ 5 g (accurate to 0.001 g) of`evenly mixed alcohol specimen in an
evaporatingdish, heat it on a 60 'C water bath for 30 minutes, use 10 ml of water to rinse the
residue in theevaporating dish in three times, and combine the washing fluids and transfer
them to a 15 mlcentrifuge tube. On a vortex mixer, oscillate it for 3 minutes, perform
ultrasonic extraction fol20 minutes, then, at 8,500 r/min, centrifuge for 5 minutes. Use 5 ml of
water to repeat theextraction once, combine the extracting solutions and reserve the
supernatant for purification.

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5.2.2 Vinegar, Soy Sauce,Sauce and Sauce Products
5.2.2.1 Weigh-take 2 g~5g (accurate to 0.001 g) of evenly mixed specimen in a 50
mIcentrifuge tube, add 1.0 g ofneutral alumina and add 5 ml ofwater. On a vortex mixer,
oscillateit for 3 minutes, then, add 15 ml of methanol, and continue to oscillate for 30 s,
performultrasonic extraction for 20 minutes; at 3,000 r/min, centrifuge for 10 minutes, and
transfer thesupernatant into the 50 mL centrifuge tube. Add the precipitate to 5.0 ml of
methanol-watersolution (75 + 25), use a glass rod to evenly stir it, then, oscillate it on a
vortex mixer for 30 s.At 3,000 r/min, centrifuge for 10 min, repeat the extraction twice, and
combine the supernatants.
5.2.2.2 Transfer all the supernatants to a separatory funnel, add 30 mL ofn-hexane, shake it
for2 minutes, and let it stand for 20 minutes for stratification. Transfer the lower aqueous
phase toan evaporating dish and evaporate it on the boiling water bath. When the liquid in
theevaporating dish is about 1 mL, use 9 mL of water to rinse the evaporating dish in three
timesand combine the washing fluids and transfer them to a 15 mL centrifuge tube.
Conductultrasonication for 5 minutes; at 3,000 r/in, centrifuge for 10 minutes and reserve it
forpurification.
5.2.3 Jelly, candies and candied fruits specimens
Weigh-take 2 g~ 5 g (accurate to 0.001 g) of pulverized and evenly mixed specimen in a
50mL centrifuge tube and add 5 mL of water. On a vortex mixer, oscillate it for 3 minutes,
then.add 15 mL ofmethanol, 0.50 mL ofzinc acetate solution and 0.50 mL of potassium
ferrocyanidesolution, continue the oscillation for 30 s. Then, in a 60 'C water bath, heat it for
15 minutesDuring the water bath, shake and disperse it, and the following steps shall be
successivelyhandled, starting from “'perform ultrasonic extraction for 20 minutes;, at 3,000
r/min, centrifugefor 10 minutes” in 5.2.2.1, till “conduct ultrasonication for 5 minutes, at 3,000
r/min, centrifugefor 10 minutes and reserve it for purification” in 5.2.2.2.

5.2.4 Other specimens

Weigh-take 2 g~ 5 g (accurate to 0.001 g) of evenly mixed specimen into a 50 mL
centrifugetube and add 5 ml of water. On a vortex mixer, oscillate for 3 minutes, then add 15
ml ofmethanol, 0.50 ml of zinc acetate solution and 0.50 mL of potassium ferrocyanide
solution.The following steps shall be successively handled, starting firom “continue to
oscillate for 30 s,perform ultrasonic extraction for 20 minutes; at 3,000 r/min, centrifuge for
10 minutes” in5.2.2.1, till “conduct ultrasonication for 5 minutes; at 3,000 r/min, centrifuge for
10 minutesand reserve it for purification’ in 5.2.2.2.

5.3 Specimen Purification

5.3.1 Differential detector
Before use, the solid phase extraction column is successively activated with 4 mL of
methanoland 4 mL of water to maintain the column moist.
Transfer all the specimen extraction supernatants into the activated solid phase
extractioncolumn and control the liquid flow rate to no more than 1 drop/s. When the liquid
level on thecolumn is about 2 mm, add 1 ml of water, and continue to maintain the liquid flow
rate at 1drop/s. After the liquid in the column is completely discharged, use 3 ml of methanol

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to eluteit, collect all the eluent and evaporate it to dryness on a boiling water bath. Use 1.00
ml ofwater to dissolve the residue (ifthe solution is turbid, it can be transferred into a
centrifuge tube.at 10,000 r/min, centrifuge for 5 minutes), filter it through a 0.45 um
hydrophilic microporousfilter membrane. The filtrate is the prepared specimen solution to be
tested.
5.3.2 Evaporative light-scattering detector
Before use, the solid phase extraction column is successively activated with 4 mL of
methanoland 4 ml of water to maintain the column moist.
Transfer all the specimen extraction supernatants into the activated solid phase
extractioncolumn and control the liquid flow rate to no more than 1 drop/s. When the liquid
level on thecolumn is about 2 mm, add 1 ml of water, and continue to maintain the liquid flow
rate at 1drop/s. After the liquid in the column is completely discharged, use 3 mL of methanol
to eluteit, collect all the eluent and evaporate it to dryness on a boiling water bath. Use 1.00
mL ofwater-acetonitrile solution (89 + 11) to dissolve the residue (if the solution is turbid, it
can betransferred into a centrifuge tube, at 10,000 r/min, centrifuge for 5 minutes), filter it
through a0.45 um hydrophobic microporous filter membrane. The filtrate is the prepared
specimensolution to be tested.
NO'TE: the supernatant of jelly samples after extraction needs to be heated in a 50 °C water
bath.then, pass through the column while it is still hot, otherwise, the extraction column will
beeasily blocked.
5.4 Blank Test
The pre-treatment of different specimens requires a blank test of the specimens to
besimultaneously conducted.
5.5 Reference Conditions of Instruments
5.5.1 Differential detector
5.5.1.1 Chromatographic column: Ci (250 mm x 4.6 mm, 5 um), or equivalent column.
5.5.1.2 Mobile phase: methanol-0.1% dipotassium hydrogen phosphate solution (20 + 80).
5.5.1.3 Flow rate: 1.0 mL/min.
5.5.1.4 Column temperature: 35°C.
5.5.1.5 Detector cell temperature: 35°C.
5.5.1.6 Sensitivity: 16.
5.5.1.7 Injection volume: 20 μL.
5.5.2 Evaporative Light Scattering Detector
5.5.2.1 Chromatographic column: C₁₈ (250 mm×4.6 mm, 5 μm) or equivalent.
5.5.2.2 Mobile phase: A is water, B is acetonitrile, water+acetonitrile=89+11.
Note: For complex sample matrices where strongly retained substances interfere with
subsequent detection, a gradient elution program can be used (see Appendix A).
5.5.2.3 Flow rate: 1.0 mL/min.

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5.5.2.4 Column temperature: 35°C.
5.5.2.5 Conditions of the evaporative light-scattering detector: set in accordance with
therequirements of different brands of evaporative light-scattering detectors under high
aqueousmobile phase conditions. For example, the atomization pressure is 0.137 MPa; the
gain is 10:the evaporation temperature is 60 oC. Or those with equivalent performance.
5.5.2.6 Injection volume: 20 μL.
5.6 Preparation of Standard Curve
5.6.1 Differential detector
In accordance with the instrument reference conditions, determine the sucralose standard
seriesof working solutions to obtain the chromatographic peak area of the corresponding
standardseries of working solutions. Take the mass concentration of the standard series of
workingsolutions as the x-coordinate and the peak area response value as the y-coordinate
to draw aworking curve. For the chromatogram of sucralose standard solution, see Figure
B.1 inAppendix B.
5.6.2 Evaporative light-scattering detector
In accordance with the instrument reference conditions, determine the sucralose standard
seriesof working solutions to obtain the chromatographic peak area of the corresponding
standardseries of working solutions. Take the mass concentration of the standard series of
workingsolutions as the x-coordinate and the peak area response value as the y-coordinate
to draw alogarithmic working curve. For the chromatogram of sucralose standard solution,
see FigureB.2.
5.7 Determination of Sample Solution
Inject 20 μL of the sample solution and blank sample solution into the liquid chromatograph
to obtain the peak area. Determine the mass concentration of sucralose in the test solution
using the standard curve.
Note: If the concentration of sucralose in the sample solution exceeds the linear range of the
standard curve, dilute it before determination.
GB 5009.298-2023

6 Expression of Analytical Results

The content of sucralose in the sample is calculated using formula (1):
X=\frac{(\rho-\rho_0) \times V \times 1000}{m \times 1000} \times f\quad\quad(1)
Where:
X – content of sucralose in the sample, in grams per kilogram (g/kg);
ρ – mass concentration of sucralose in the sample solution obtained from the standard curve,
in milligrams per milliliter (mg/mL);
ρ₀ – mass concentration of sucralose in the sample blank obtained from the standard curve,
in milligrams per milliliter (mg/mL);

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V – volume of the sample solution, in milliliters (mL);
1000 – conversion factor;
m – mass of the sample, in grams (g);
f – dilution factor.
Results are retained to 3 significant figures.

7 Precision
The absolute difference between two independent determination results obtained under
repeatability conditions shall not exceed 10% of the arithmetic mean.

8 Others
For distilled wine: When the sample size is 2.000 g and the volume is fixed to 1.00 mL, the
detection limit of this method is 0.0075 g/kg, and the quantification limit is 0.02 g/kg.
For other foods: When the sample size is 2.000 g and the volume is fixed to 1.00 mL, the
detection limit of this method is 0.01 g/kg, and the quantification limit is 0.03 g/kg.
Second Method: High Performance Liquid Chromatography-Tandem Mass Spectrometry

9 Principle
Sucralose in the sample is extracted with water, proteins and fats are removed, purified by a
solid-phase extraction column, detected by a high performance liquid chromatography-
tandem mass spectrometer, and quantified by the external standard method.

10 Reagents and Materials

Unless otherwise specified, all reagents used in this method are of analytical purity, and
water conforms to grade I water specified in GB/T 6682.
10.1 Reagents
10.1.1 Methanol (CH₃OH).

10.1.2 Methanol (CH₃OH): chromatographically pure.

10.1.3 n-Hexane (C₆ H₁₄).
10.1.4 Zinc acetate dihydrate [Zn(CH₃COO)₂·2H₂O].
10.1.5 Potassium ferrocyanide trihydrate [K₄Fe(CN)₆ ·3H₂O].
10.2 Reagent Preparation
10.2.1 Zinc acetate solution (219 g/L): Weigh 21.9 g of zinc acetate, add 3 mL of acetic acid,
and dissolve in water to make 100 mL.

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10.2.2 Potassium ferrocyanide solution (106 g/L): Weigh 10.6 g of potassium ferrocyanide,
and dissolve in water to make 100 mL.
10.3 Standard Substance
Sucralose (CAS No.: 56038-13-2) with a purity of ≥99%, or a standard substance certified by
the state and issued with a standard substance certificate.
10.4 Preparation of Standard Solutions
10.4.1 Sucralose standard stock solution (10.0 mg/mL): Weigh 0.25 g of sucralose standard
substance (accurate to 0.0001 g), dissolve in water, transfer to a 25 mL volumetric flask,
dilute to the mark, and mix well. Store the stock solution in a refrigerator at 4°C with a shelf
life of 6 months.
10.4.2 Sucralose standard intermediate solution (10 mg/L): Pipette 10.0 μL of the sucralose
standard stock solution (10.0 mg/mL) into a 10 mL volumetric flask, dilute to the mark with
water, and mix well. Store in a refrigerator at 4°C with a shelf life of 3 months.
10.4.3 Sucralose standard series working solutions: Pipette 0.100 mL, 0.200 mL, 0.500 mL,
1.00 mL, and 1.50 mL of the sucralose standard intermediate solution (10 mg/L) into 10 mL
volumetric flasks respectively, dilute to the mark with water, and mix well. Prepare standard
series working solutions with mass concentrations of 0.100 mg/L, 0.200 mg/L, 0.500 mg/L,
1.00 mg/L, and 1.50 mg/L. Prepare fresh before use.
10.5 Materials
10.5.1 Solid-phase extraction column (specification: 200 mg/6 mL, packed with N-
vinylpyrrolidone and divinylbenzene hydrophilic-lipophilic balanced filler or equivalent).
10.5.2 0.22 μm hydrophilic microporous membrane and 0.22 μm hydrophobic microporous
membrane.

11 Instruments and Equipment

11.1 High performance liquid chromatography-tandem mass spectrometer: equipped with an
electrospray ion source.
11.2 Balance: with sensitivities of 0.1 mg and 1 mg respectively.
11.3 Vortex mixer.
11.4 Water bath.
11.5 Ultrasonic generator: 50 kHz.
11.6 Centrifuge: rotational speed ≥8500 r/min.
11.7 Solid-phase extraction device.
11.8 Homogenizer.
11.9 Crusher.

12 AnalyticalProcedures

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12.1 Specimen Preparation
For liquid samples: shake them well; for semi-solid samples and powdery samples with
uniformmatrix: directly determine them; other samples need to be homogenized or evenly
pulverized.
12.2 Specimen Extraction
12.2.1 Alcoholic Specimen
12.2.2 Jelly, candies and candied fruits specimens
Weigh-take 2 g (accurate to 0.001 g) of specimen in a 50 mL centrifuge tube and add 10 mL
ofwater. On a vortex mixer, oscillate it for 3 minutes, then, add 1.00 mL of zinc acetate
solutionand 1.00 ml of potassium ferrocyanide solution. In a 60 'C water bath, heat it for 15
minutesDuring the water bath, pay attention to shake and disperse it. Perform ultrasonic
extraction for20 minutes; at 8,500 r/min, centrifuge for 5 minutes. Take the supernatant (if
necessary, use afast quantitative filter paper to filter it) in a 25 mL, volumetric flask, use 10
mL of water torepeat the extraction once. After combining the extracting solutions, add water
to reach aconstant volume to the scale. Evenly mix it, then transfer it to a 50 ml centrifuge
tube. Add 15mL of n-hexane, on a vortex mixer, oscillate it for 3 minutes. At 8,500 r/min,
centrifuge for 5minutes. Discard the upper n-hexane layer and reserve the subnatant for
purification.
12.2.3 Other specimens
Weigh 2 g of the sample (accurate to 0.001 g) into a 50 mL centrifuge tube, add 10 mL of
water, vortex for 3 min, add 1.00 mL of zinc acetate solution and 1.00 mL of potassium
ferrocyanide solution, ultrasonically extract for 20 min, centrifuge at 8500 r/min for 5 min,
take the supernatant (filter with rapid quantitative filter paper if necessary) into a 25 mL
volumetric flask, re-extract with 10 mL of water once, combine the extracts, dilute to the
mark with water, mix well, transfer to a 50 mL centrifuge tube, add 15 mL of n-hexane,
vortex for 3 min, centrifuge at 8500 r/min for 5 min, discard the upper n-hexane layer, and
the lower clear liquid is to be purified.
12.3 Specimen Purification
Before use, the solid phase extraction column is successively activated with 4 mL of
methanoland 4 mL of water to maintain the column moist.
Accurately transfer-take 10.00 mL of the specimen extracting solution into the activated
solidphase extraction column and control the liquid flow rate to no more than 1 drop/s. After
theliquid in the column is completely discharged, use 3 mL of methanol to elute it, collect
themethanol eluent in a 10 m volumetric flask, add water to reach a constant volume to the
scaleand evenly mix it. Use a 0.22 um hydrophobic microporous filter membrane to filter it.
Thefiltrate is the prepared specimen solution to be tested.
NO'TE: the solution of jelly samples to be purified needs to be heated in a 50 °C water bath,
then,pass through the column while it is still hot, otherwise, the extraction column will be
easilyblocked.
12.4 Blank Test

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The pre-treatment of different specimens requires a blank test of the specimens to
besimultaneously conducted.
12.5 Instrument Reference Conditions
12.5.1 Liquid Chromatography Conditions
12.5.1.1 Chromatographic column: C₁₈ (100 mm×4.6 mm, 2.5 μm) or equivalent.
12.5.1.2 Mobile phase: A is water, B is methanol, with gradient elution program as shown in
Table 1.
Table 1 Gradient Elution Program

Time/min A (Water) (volume B (Methanol) (volume

fraction)/% fraction)/%

0 85 15

3 10 90

5 10 90

5.1 85 15

7 85 15

12.5.1.3 Flow rate: 0.6 mL/min.

12.5.1.4 Column temperature: 40°C.
12.5.1.5 Injection volume: 1 μL.
12.5.2 Mass Spectrometry Conditions
12.5.2.1 Ionization mode: electrospray negative ion mode.
12.5.2.2 Monitoring mode: multiple reaction monitoring (MRM).
12.5.2.3 Capillary voltage: 3.0 kV.
12.5.2.4 Desolvation gas temperature: 550°C.
12.5.2.5 Desolvation gas flow rate: 1000 L/h.
12.5.2.6 Ion source temperature: 150°C.
12.5.2.7 Qualitative ion pairs, quantitative ion pairs, cone voltage, and collision voltage are
shown in Table 2.

Table 2 Qualitative Ion Pairs, Quantitative Ion Pairs, Cone Voltage, and Collision
Voltage of Sucralose

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Chinese Name Qualitative Ion Quantitative Ion Cone Voltage/V Collision
Pairs (m/z) Pairs (m/z) Voltage/V
(Parent (Parent
Ion/Daughter Ion/Daughter
Ion) Ion)

Sucralose 394.8/359.0 394.8/359.0 -50 17

396.8/361.0 - -50 17

12.5.3 Qualitative Confirmation

Determine the sample solution and standard working solution under the instrument reference
conditions. If the retention time of the sucralose mass chromatographic peak in the sample is
consistent with that in the standard working solution (variation within ±2.5%); and all
monitored qualitative ions in the sample are present, and the relative abundance ratio (k) of
the two daughter ions of sucralose compared with that of the standard working solution with
equivalent concentration has an allowable deviation not exceeding the range specified in
Table 3, then sucralose is judged to be present in the sample.

Table 3 Maximum Allowable Deviation of Relative Ion Abundance in Qualitative

Confirmation

Relative Ion k > 50 20 < k ≤ 50 10 < k ≤ 20 k ≤ 10

Abundance/%

Maximum Allowable ±20 ±25 ±30 ±50

Deviation/%

See Figure C.1 in Appendix C for the liquid chromatography-mass spectrometry chart of the
sucralose standard solution.
12.5.4 Quantitative Determination
12.5.4.1 Preparation of Working Curve
Determine the sucralose standard series working solutions under the instrument reference
conditions to obtain the corresponding mass chromatographic peak areas. Plot a working
curve with the mass concentration of the standard series working solutions as the abscissa
and the mass chromatographic peak area as the ordinate.
12.5.4.2 Determination of Sample Solution

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Determine the sample solution under the instrument reference conditions to obtain the
corresponding mass chromatographic peak area. Determine the mass concentration of
sucralose in the sample solution using the working curve.
Note: If the concentration of sucralose in the sample solution exceeds the linear range of the
standard curve, dilute it before determination.

13 Expression of Analytical Results

The content of sucralose in distilled wine samples is calculated using formula (2), and that in
other samples using formula (3):
X=\frac{(\rho-\rho_0) \times V}{m \times 1000} \times f\quad\quad(2)
Where:
X – content of sucralose in the sample, in grams per kilogram (g/kg);
ρ – mass concentration of sucralose in the sample solution obtained from the working curve,
in milligrams per liter (mg/L);
ρ₀ – mass concentration of sucralose in the sample blank obtained from the working curve,
in milligrams per liter (mg/L);
V – volume of the sample solution, in milliliters (mL);
m – mass of the sample, in grams (g);
1000 – conversion factor;
f – dilution factor.
X=\frac{(\rho-\rho_0) \times V_1 \times V_3}{m \times 1000 \times V_2} \times f\quad\quad(3)
Where:
X – content of sucralose in the sample, in grams per kilogram (g/kg);
ρ – mass concentration of sucralose in the sample solution obtained from the working curve,
in milligrams per liter (mg/L);
ρ₀ – mass concentration of sucralose in the sample blank obtained from the working curve,
in milligrams per liter (mg/L);
V₁ – volume of the sample extract, in milliliters (mL);
V₃ – final volume of the sample after purification and elution, in milliliters (mL);
m – mass of the sample, in grams (g);
1000 – conversion factor;
V₂ – volume of the sample aliquot for purification, in milliliters (mL);
f – dilution factor.
Results are retained to 3 significant figures.

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14 Precision
The absolute difference between two independent determination results obtained under
repeatability conditions shall not exceed 10% of the arithmetic mean.

15 Others
For distilled wine: When the sample size is 2.000 g and the volume is fixed to 10.00 mL, the
detection limit of this method is 0.0003 g/kg, and the quantification limit is 0.0006 g/kg.
For other foods: When the sample size is 2.000 g and the volume is fixed to 25.00 mL, the
detection limit of this method is 0.001 g/kg, and the quantification limit is 0.003 g/kg.
Appendix A Liquid Chromatography Elution Program (for Evaporative Light Scattering
Detector)

The liquid chromatography gradient elution program for evaporative light scattering detectors
is shown in Table A.1.

Table A.1 Liquid Chromatography Gradient Elution Program

Time/min A (Water) (volume B (Acetonitrile) (volume

fraction)/% fraction)/%

0 89 11

14 89 11

15 10 90

22 10 90

23 89 11

26 89 11

Appendix B
Liquid Chromatograms of Sucralose Standard Solutions
B.1 The liquid chromatogram of the sucralose standard solution (refractive index detector) is
shown in Figure B.1.

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Figure B.1 Liquid Chromatogram of 0.400 mg/mL Sucralose Standard Solution (Refractive Index Detector)

B.2 The liquid chromatogram of the sucralose standard solution (evaporative light scattering
detector) is shown in Figure B.2.

Figure B.2 Liquid Chromatogram of 0.400 mg/mL Sucralose Standard Solution (Evaporative
Light Scattering Detector)
Appendix C
Liquid Chromatography-Mass Spectromogram of Sucralose Standard Solution
The liquid chromatography-mass spectromogram of the sucralose standard solution is
shown in Figure C.1.

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Figure C.1 Liquid Chromatography-Mass Spectromogram of 0.100 mg/L Sucralose Standard
Solution

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