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GB 5009.8-2023: National food safety standard - Determination of fructose, glucose, sucrose, maltose and lactose in foods
GB 5009.8-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standard - Determination of fructose,
glucose, sucrose, maltose and lactose in foods
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 apparatuses... 6
5 Analysis steps... 6
6 Expression of analysis results... 8
7 Precision... 9
8 Others... 9
9 Principle... 9
10 Reagents and materials... 9
11 Instruments and apparatuses... 11
12 Test steps... 11
13 Expression of analysis results... 14
14 Precision... 14
15 Others... 14
16 Principle... 15
17 Reagents and solutions... 15
18 Instruments and apparatuses... 16
19 Analysis steps... 16
20 Expression of analysis results... 19
21 Precision... 21
22 Principle... 21
23 Reagents and materials... 21
24 Instruments and apparatuses... 22
25 Analysis steps... 22
26 Expression of analysis results... 24
27 Precision... 25
Appendix A HPLC chromatograms of standard solutions of fructose, glucose, sucrose,
maltose and lactose... 26
Appendix B Ion chromatogram of standard solutions of fructose, glucose, sucrose,
maltose and lactose... 28
National food safety standard
Determination of fructose, glucose, sucrose, maltose and
lactose in foods
1 Scope
This Standard specifies the methods for the determination of fructose, glucose, sucrose,
maltose and lactose in foods.
Method 1 - High performance liquid chromatography applies to the determination of
fructose, glucose, sucrose, maltose and lactose in grains and grain products, milk and
dairy products, fruits, vegetables and fruit and vegetable products, sweeteners, candies,
beverages and infant foods.
Method 2 - Ion chromatography applies to the determination of fructose, glucose,
sucrose, maltose, and lactose in foods.
Method 3 - Acid hydrolysis-Rhein-Eynon method applies to the determination of
sucrose in foods.
Method 4 - Rhine-Enon method applies to the determination of lactose in infant foods
and dairy products.
Method 1 - High performance liquid chromatography
2 Principle
The fructose, glucose, sucrose, maltose and lactose in the sample are extracted, and then
separated by high performance liquid chromatography column, detected by a
differential refractive index detector or an evaporative light scattering detector, and
quantified by the external standard method.
3 Reagents and materials
Unless otherwise specified, all the reagents in this method are analytical reagents, and
the water is grade-1 water specified by GB/T 6682.
3.1 Reagents
3.1.1 Acetonitrile (C2H3N). chromatographic purity.
3.1.2 Zinc acetate [Zn(CH3COO)2·2H2O].
3.1.3 Potassium ferrocyanide [K4Fe(CN)6·3H2O].
3.1.4 Glacial acetic acid (CH3COOH).
3.2 Preparation of reagents
3.2.1 Zinc acetate solution (1 mol/L). Weigh 21.9 g of zinc acetate; add 3 mL of glacial
acetic acid; dissolve in water and dilute to 100 mL; mix well.
3.2.2 Potassium ferrocyanide solution (0.25 mol/L). Weigh 10.6 g of potassium
ferrocyanide; dissolve in water and dilute to 100 mL; mix well.
3.3 Standards
3.3.1 Fructose (C6H12O6, CAS number. 57-48-7). purity ≥99%, or standard material
certified by the country and awarded a standard material certificate.
3.3.2 Glucose (C6H12O6, CAS number. 50-99-7). purity ≥99%, or standard material
certified by the country and awarded a standard material certificate.
3.3.3 Sucrose (C12H22O11, CAS number. 57-50-1). purity ≥99%, or standard material
certified by the country and awarded a standard material certificate.
3.3.4 Maltose (C12H22O11, CAS number. 69-79-4). purity ≥99%, or standard material
certified by the country and awarded a standard material certificate.
3.3.5 Lactose (C12H22O11, CAS number. 63-42-3). purity ≥99%, or standard material
certified by the country and awarded a standard material certificate.
3.4 Preparation of standard solutions
3.4.1 Mixed standard stock solution (20.0 mg/mL). Respectively weigh 1 g (accurate
to 0.001 g) each of fructose dried at 90 °C ± 2 °C for 2 h and glucose, sucrose, maltose
and lactose dried at 96 °C ± 2 °C for 2 h; dissolve in water and then transfer to the 50
mL volumetric flasks; add 2.5 mL of acetonitrile; use water to adjust the volume to the
mark. Store at 0 °C ~ 4 °C and seal, with a shelf life of 3 months.
3.4.2 Mixed standard working solution. Take 0.100 mL, 1.00 mL, 2.00 mL, 3.00 mL
and 5.00 mL of mixed standard stock solution (20.0 mg/mL) into the 10.0 mL
volumetric flask; use water to dilute to the mark, to prepare the mixed standard working
solutions of fructose, glucose, sucrose, maltose and lactose with mass concentrations of
0.200 mg/mL, 2.00 mg/mL, 4.00 mg/mL, 6.00 mg/mL and 10.0 mg/mL. The
concentration of the mixed standard working solution can be appropriately adjusted
according to the concentration of the actual sample solution. Prepare when necessary.
3.5 Materials
Weigh 50 g (accurate to 0.01 g) of the mixed sample in an evaporating dish; stir gently
on a water bath to remove gas and alcohol; transfer to a 100 mL volumetric flask after
cooling; slowly add 5 mL of zinc acetate solution and 5 mL of potassium ferrocyanide
solution; use water to adjust the volume to the mark; mix well; let stand.
5.1.2.3 Syrup and honey samples
Weigh 1 g ~ 2 g (accurate to 0.001 g) of the mixed sample into a 100 mL colorimetric
tube; add about 50 mL of water; vortex and mix until fully dissolved; transfer to a 100
mL volumetric flask; use water to fix the volume to the mark; mix well; let stand.
5.1.2.4 Other samples
Weigh 1 g ~ 10 g (accurate to 0.001 g) of the crushed or mixed sample (10 g when the
target sugar content is ≤5%; 5 g when the content is 5% ~ 10%; 2 g when the content is
10% ~ 40%; 1 g when the content is ≥40%) into a 100 mL colorimetric tube; add about
50 mL of water; then, slowly add 5 mL of zinc acetate solution and 5 mL of potassium
ferrocyanide solution; vortex and mix, and sonicate for 30 minutes; transfer to a 100
mL volumetric flask; use water to adjust the volume to the mark; mix well; let stand.
5.1.3 Purification
For the above sample extract solution, use a filter paper to filter (discard the primary
filtrate) or centrifuge to obtain the supernatant; then, use a 0.45 μm water-based
membrane syringe to filter into a sample bottle for analysis by a high-performance
liquid chromatograph.
5.2 Apparatus reference conditions
Apparatus reference conditions are as below.
a) Chromatographic column. Amino chromatographic column (4.6 mm × 250 mm,
particle size 5 μm, aminosilane bonded silica gel as filler), or one with equivalent
performance;
b) Mobile phase. acetonitrile + water = 70 + 30 (volume ratio);
c) Flow velocity. 1.0 mL/min;
d) Column temperature. 40 ℃;
e) Injection volume. 10 μL;
f) Differential refractive index detector conditions. temperature 40 °C;
g) Evaporative light scattering detector conditions. drift tube temperature 80 °C ~
90 °C; nitrogen flow rate 2.5 L/min.
5.3 Preparation of the standard curve
Inject the mixed standard working solution into the high-performance liquid
chromatograph in order from low to high concentration, and measure the corresponding
peak areas or peak heights of fructose, glucose, sucrose, maltose a...