GB/T 30390-2026Determination of fructose, glucose, sucrose, stachyose and raffinose in oilseeds (English PDF)

4.2.1 Reagents

4.2.1.1 Acetonitrile (CH3CN). chromatographic grade.

4.2.1.2 Acetic acid (CH3COOH).

4.2.1.3 Petroleum ether. boiling range 30°C~60°C.

4.2.1.4 Zinc acetate dihydrate [Zn(CH3COO)2·2H2O].

4.2.1.5 Potassium ferrocyanide trihydrate [K4Fe(CN)6·3H2O].

4.2.2.1 Zinc acetate solution (1 mol/L). Weigh

21.9 g of zinc acetate dihydrate (4.2.1.4), add 3 mL of acetic acid (4.2.1.2), and dissolve in water. Make up to 100 mL.

4.2.2.2 Potassium ferrocyanide solution (1 mol/L). Weigh

10.6 g of potassium ferrocyanide trihydrate (4.2.1.5), add water to dissolve and make up to 100 mL.

4.2.3 Standard Samples/Standard Substances

4.2.3.1 Fructose (C6H12O6, CAS No.. 57-48-7). Purity >= 99%.

4.2.3.2 Glucose (C6H12O6, CAS No.. 50-99-7). Purity >= 99%.

4.2.3.3 Sucrose (C12H22O12, CAS No.. 57-50-1). Purity >= 99%.

4.2.3.4 Stachyose (C24H42O21, CAS No.. 54261-98-2). Purity >= 98%.

4.2.3.5 Raffinose (C18H32O16, CAS No.. 512-69-6). Purity >= 99%.

4.2.4.1 Standard stock solution (20 mg/mL). Weigh 1 g (accurate to

0.1 mg) of fructose (4.2.3.1), glucose (4.2.3.2), and sucrose respectively. (4.2.3.3), stachyose (4.2.3.4) and raffinose (4.2.3.5) standard samples/standard substances were dissolved in water and diluted to volume in a 50 mL volumetric flask. The solution should be stored at 4°C away from light and has a shelf life of 1 month.

4.2.4.2 Standard working solutions. Measure

0.05 mL,

0.25 mL,

0.50 mL,

5.00 mL of standard stock solution, respectively. (4.2.4.1) Dilute to volume in a 10 mL volumetric flask to prepare a mixed standard working solution. The mixed standard working solution contains fructose, glucose, sucrose, and... The mass concentrations of stachyose and raffinose were

0.1 mg/mL,

0.5 mg/mL,

5.0 mg/mL, respectively.

10.0 mg/mL. Prepare fresh before use.

4.2.5 Materials

4.2.5.1 Aqueous filter membrane needle filter. 0.22µm.

4.2.5.2 Qualitative rapid filter paper.

4.3 Instruments and Equipment

4.3.1 High-performance liquid chromatograph. equipped with a differential refractive index detector or an evaporative light scattering detector.

4.3.2 Electronic balance. sensitivity

0.1 mg and 1 mg.

4.3.3 Constant temperature drying oven.

4.3.4 Vortex mixer.

4.3.5 Crusher. Rotation speed not less than 12000 r/min.

4.3.6 Constant temperature water bath. 60°C±2°C.

4.3.7 Centrifuge. rotation speed not less than.2000 r/min.

4.4 Sample Preparation

4.4.1 Take about.200g of sample according to GB 5491, crush it with a pulverizer, pass it through a 0.64mm sieve, and place it in a sealed container.

4.4.2 Peanut samples should be sliced first and then crushed using a grinder.

4.5 Test Procedure

4.5.1 Sample Extraction Weigh 5g of the sample (accurate to 1mg), place it in a 50mL stoppered centrifuge tube, add 25mL of petroleum ether (4.2.1.3), vortex for 1min. Centrifuge at.2000 rpm for 5 minutes, discarding the petroleum ether layer. Repeat the above steps three times. Evaporate the remaining petroleum ether in a 60°C water bath, then rinse with a glass filter. Transfer the sample to a 100 mL Erlenmeyer flask using a glass rod. Rinse the centrifuge tube twice with 30 mL of water, and add the washings to the Erlenmeyer flask. Slowly add acetic acid. Add 5 mL each of zinc solution (4.2.2.1) and potassium ferrocyanide solution (4.2.2.2), vortex at.2000 rpm for 30 min at room temperature, and extract. Transfer to... Dilute to the mark with water in a 100 mL volumetric flask and mix well. Filter using qualitative rapid filter paper (4.2.5.2), and filter the filtrate using a water-based filter membrane syringe filter. Filter using the apparatus (4.2.5.1), and the filtrate is to be tested.

4.5.2 Reference conditions for high performance liquid chromatography The reference conditions for high performance liquid chromatography are as follows:

a) Chromatographic column. amino column (

4.6 mm × 250 mm, particle size 5 µm), or equivalent;

b) Column temperature. 40°C;

c) Mobile phase. Acetonitrile (4.2.1.1) Water = 75 25 (V/V);

e) Injection volume. 10 µL.

4.5.3 Plotting the Standard Curve Following the high-performance liquid chromatography (HPLC) working conditions set in 4.5.2, fructose, glucose, sucrose, and water in the standard working solution (4.2.4.2) were determined sequentially. Chromatographic peak areas of threose and raffinose. The peak areas of fructose, glucose, sucrose, raffinose, and stachyose in the standard working solution (4.2.4.2) were determined by mass concentration. Plot standard curves for the peak areas of fructose, glucose, sucrose, stachyose, and raffinose, with altitude as the x-axis and peak area as the y-axis. The high-performance liquid chromatograms of the quasi-samples/standards are shown in Figure A.1 in Appendix A.

4.5.4 Determination of Sample Solution The test solution was pipetted into a sample vial and analyzed using a high-performance liquid chromatograph (HPLC) equipped with a differential refractive index detector or an evaporative light detector, according to the standard curve. Calculate the content of fructose, glucose, sucrose, stachyose, and raffinose in the sample solution, or use regression equations to calculate the content of fructose, glucose, and raffinose in the sample solution. The content of sucrose, stachyose, and raffinose. If the sugar content in the sample exceeds the concentration of the standard curve, an appropriate dilution factor should be selected, and the sample should be diluted with water.

4.5.5 Blank Test The determination was performed using the exact same analytical procedure, except that no sample was added.

4.6 Determination of Moisture and Volatile Matter Content The moisture and volatile matter content in the sample shall be determined according to GB/T 14489.1.

4.7 Data Processing The contents of fructose, glucose, sucrose, stachyose and raffinose in the sample were calculated according to the standard curve or formula (1).

4.8 Precision

4.8.1 Repeatability Under repeatability conditions, the absolute difference between two independent test results should not exceed 10% of the arithmetic mean.

4.8.2 Reproducibility Under reproducibility conditions, the absolute difference between two independent test results is no greater than 10% of the arithmetic mean.

4.9 Other When the sample weight is 5g, the method detection limits for fructose, glucose, sucrose, and raffinose are all 2mg/g, and the method detection limit for stachyose is [missing value]. 5mg/g.

5.2.1 Reagents

5.2.1.1 Sodium hydroxide solution (50%), chromatographic grade, for ion chromatography.

5.2.1.2 Acetic acid (CH3COOH).

5.2.1.3 Petroleum ether. boiling range 30°C~60°C.

5.2.1.4 Zinc acetate dihydrate [Zn(CH3COO)2·2H2O].

5.2.1.5 Potassium ferrocyanide trihydrate [K4Fe(CN)6·3H2O].

5.2.2.1 Zinc acetate solution (1 mol/L). Weigh

21.9 g of zinc acetate dihydrate (5.2.1.4), add 3 mL of acetic acid (5.2.1.2), and dissolve in water. Make up to 100 mL.

5.2.2.2 Potassium ferrocyanide solution (1 mol/L). Weigh

10.6 g of potassium ferrocyanide trihydrate (5.2.1.5), dissolve in water, and dilute to a final volume. 100mL.

5.2.2.3 Sodium hydroxide solution (200 mmol/L). Measure

10.5 mL of 50% sodium hydroxide solution (5.2.1.1), and rinse with pre-degassed water. Dilute and bring the volume to 1000 mL under nitrogen protection.

5.2.3 Standard Samples/Standard Substances

5.2.3.1 Fructose (C6H12O6, CAS No.. 57-48-7). Purity >= 99%.

5.2.3.2 Glucose (C6H12O6, CAS No.. 50-99-7). Purity >= 99%.

5.2.3.3 Sucrose (C12H22O12, CAS No.. 57-50-1). Purity >= 99%.

5.2.3.4 Stachyose (C24H42O21, CAS No.. 54261-98-2). Purity >= 98%.

5.2.3.5 Raffinose (C18H32O16, CAS No.. 512-69-6). Purity >= 99%.

5.2.4 Preparation of Standard Solutions

5.2.4.1 Standard stock solutions (10 mg/mL). Weigh out fructose (5.2.3.1), glucose (5.2.3.2), sucrose (5.2.3.3), and stachyose respectively. Dissolve 100.0 mg each of (5.2.3.4) and raffinose (5.2.3.5) in a 10 mL volumetric flask, and dilute to volume with water. Store the solution at 4 °C in the dark. Stored, valid for one month.

5.2.4.2 Standard Intermediate Solution (

1.0 mg/mL). Pipette

1.0 mL of the standard stock solution (5.2.4.1) into a 10 mL volumetric flask and dilute with water. Release and dilute to a final volume to prepare a standard intermediate solution with a mass concentration of

5.2.4.3 Standard working solution. Measure

0.02 mL,

0.05 mL,

0.10 mL,

0.20 mL,

0.50 mL,

2.00 mL of standard working solution respectively. The intermediate solution (5.2.4.2) was diluted to volume in a 10 mL volumetric flask to prepare a mixed standard working solution. The mass concentrations of the five sugars in the standard working solution were then determined. The concentrations are 2 mg/L, 5 mg/L, 10 mg/L, 20 mg/L, 50 mg/L, 100 mg/L, and.200 mg/L, respectively. Prepare immediately before use.

5.2.5 Materials

5.2.5.1 Aqueous membrane needle filter. 0.22µm.

5.2.5.2 Qualitative rapid filter paper.

5.3 Instruments and Equipment

5.3.1 Ion chromatograph. equipped with gradient pump, pulse amperometric detector, and Au working electrode.

5.3.2 Electronic balance. sensitivity

0.1 mg and 1 mg.

5.3.3 Constant temperature drying oven.

5.3.4 Vortex mixer.

5.3.5 Crusher. Rotation speed not less than 12000 r/min.

5.3.6 Constant temperature water bath. 60°C±2°C.

5.3.7 Centrifuge. rotation speed not less than.2000 r/min.

5.4 Sample Preparation

5.4.1 Take approximately.200g of sample according to GB 5491, crush it using a pulverizer, pass it through a 0.64mm sieve, and place it in a sealed container.

5.4.2 Peanut samples should be sliced first and then crushed using a grinder.

5.5 Test Procedure

5.5.1 Sample Extraction Same as 4.5.1.

5.5.2 Reference conditions for ion chromatography The reference conditions for ion chromatography are as follows:

a) Chromatographic column. An anion exchange column (3mm × 100mm) filled with quaternary ammonium salt functional groups and polystyrene/divinylbenzene polymer resin. 150mm (particle size 6.5µm), or equivalent performance;

b) Column temperature. 30°C;

c) Mobile phase. Sodium hydroxide solution (5.2.2.3) + Water = 91.9 (V/V);

0.5 mL/min;

e) Injection volume. 1 µL;

f) Detection cell temperature. 35°C;

g) Detector. Pulse ampere detector, Au working electrode, Ag/AgCl reference electrode, the detector potential waveform program is shown in Table 1.

5.5.3 Plotting the Standard Curve Following the ion chromatography working conditions set in 5.5.2, fructose, glucose, sucrose, and stachyose in the standard working solution (5.2.4.3) were determined sequentially. The chromatographic peak areas of fructose and raffinose were determined based on the mass concentrations of fructose, glucose, sucrose, stachyose, and raffinose in the standard working solution (5.2.4.3). Standard curves for fructose, glucose, sucrose, stachyose, and raffinose were plotted using the corresponding peak areas. Chromatograms of standard samples/standard substances are shown in Appendix B. Figure B.1 in the text.

5.5.4 Determination of Sample Solution The sample solution was analyzed according to the ion chromatography working conditions set in 5.5.2.Qualitative analysis was performed based on retention time, and the results were obtained from the standard curve. The sugar content in the sample. If the sugar content in the sample exceeds the concentration of the standard curve, an appropriate dilution factor should be selected, and the sample should be diluted with water.

5.5.5 Blank Test The determination was performed using the exact same analytical procedure, except that no sample was added.

5.6 Determination of Moisture and Volatile Matter Content The moisture and volatile matter content in the sample shall be determined according to GB/T 14489.1.

5.7 Data Processing The contents of fructose, glucose, sucrose, stachyose and raffinose in the sample were calculated according to the standard curve or formula (2).

5.8 Precision

5.8.1 Repeatability Under repeatability conditions, the absolute difference between two independent test results should not exceed 15% of the arithmetic mean.

5.8.2 Reproducibility Under reproducibility conditions, the absolute difference between two independent test results is no greater than 20% of the arithmetic mean.

5.9 Other When the sample weight is 5g, the method detection limits for fructose, glucose, sucrose, stachyose, and raffinose are 0.026mg/g, 0.008mg/g, and 0.008mg/g, respectively. 0.018mg/g, 0.018mg/g, 0.020mg/g. GB/T 30390-2026. Determination of fructose, glucose, sucrose, stachyose and raffinose content in oilseeds ICS

33 National Standards of the People's Republic of China Replaces GB/T 30390-2013 Oilseeds contain fructose, glucose, sucrose, stachyose, and Determination of raffinose content Published on 2026-03-

31 Implemented on October 1, 2026 State Administration for Market Regulation The State Administration for Standardization issued a statement.