GB 5009.88-2023 English PDF (GB5009.88-2023)
GB 5009.88-2023 English PDF (GB5009.88-2023)
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GB 5009.88-2023: National food safety standard - Determination of dietary fiber in foods
GB 5009.88-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards -- Determination of dietary
fiber 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 Terms and definitions... 4
3 Principle... 5
4 Reagents and materials... 5
5 Instruments and equipment... 8
6 Analysis steps... 9
Annex A Definition of enzyme activity units and criteria for determining enzyme
activity... 18
Annex B Chromatogram of dietary fiber... 20
National food safety standards -- Determination of dietary
fiber in food
1 Scope
This Standard specifies the method for determination of dietary fiber in food.
This Standard is applicable to the determination of total dietary fiber, soluble dietary
fiber, and insoluble dietary fiber in plant foods and their products, as well as foods with
added dietary fiber components.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 dietary fiber; DF
Carbohydrate polymers that cannot be digested and absorbed by the human small
intestine and have a degree of polymerization ≥3.
According to its source, dietary fiber is divided into. Carbohydrate Polymers naturally
present in the edible parts of plants, such as cellulose, hemicellulose, pectin, lignin, etc.
in plant cell walls; and Carbohydrate Polymers that are isolated, extracted or synthesized
from food raw materials using physical, enzymatic or chemical means and have been
proven by scientific evidence to have beneficial physiological effects.
2.2 soluble dietary fiber; SDF
The portion of dietary fiber that is soluble in water, including indigestible
oligosaccharides and some polysaccharides.
During the detection process, based on whether it can be precipitated by 78% ethanol,
it is divided into precipitable soluble dietary fiber (SDFP) and non-precipitable soluble
dietary fiber (SDFS).
2.3 insoluble dietary fiber; IDF
The portion of dietary fiber that is insoluble in water.
2.4 total dietary fiber; TDF
The sum of soluble dietary fiber and insoluble dietary fiber.
3 Principle
The specimen is homogenized and enzymatically hydrolyzed to remove starch and
protein to obtain an indigestible enzymatic hydrolyzate.
The enzymatic hydrolyzate is precipitated with 78% ethanol. Collect the precipitated
part, wash, dry and weigh it, and then measure the mass of DF (including IDF and SDFP)
in the residue. Collect the filtrate portion. After desalting and concentration, liquid
chromatography (internal standard method) is used to determine SDFS. The sum of the
two is TDF.
The enzymatic hydrolyzate is filtered directly and washed with hot water. The filter
residue is collected, washed, dried, and weighed to determine the mass of the IDF
residue. The filtrate is collected and precipitated with 78% ethanol. The sediment is
dried and weighed. The mass of the SDFP residue is determined. The filtrate fraction is
measured as SDFS. The sum of SDFS and SDFP is SDF.
The mass of TDF, IDF and SDFP residues needs to be deducted from the mass of
residual protein, ash and reagent blank to obtain the dietary fiber content of the
corresponding part.
4 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure, and the
water is grade two water specified in GB/T 6682.
4.1 Reagents
4.1.1 95% ethanol (CH3CH2OH).
4.1.2 Acetone (CH3COCH3).
4.1.3 Petroleum ether. boiling range 30℃~60℃.
4.1.4 Sodium hydroxide (NaOH).
4.1.5 Potassium dichromate (K2Cr2O7).
4.1.6 Trishydroxymethylaminomethane (C4H11NO3, Tris).
4.1.7 Maleic acid (C4H4O4).
4.1.8 Concentrated hydrochloric acid (HCl).
4.1.9 Glacial acetic acid (C2H4O2).
4.2.4 Hydrochloric acid solution (2 mol/L). Measure 167 mL of concentrated
hydrochloric acid. Slowly add 500 mL of water. Mix well and add water to dilute to 1
L.
4.2.5 Sodium hydroxide solution (4 mol/L). Weigh 16 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water to dilute to 100 mL and mix well.
4.2.6 Sodium hydroxide solution (1 mol/L). Weigh 4 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water to dilute to 100 mL and mix well.
4.2.7 Sodium hydroxide solution (0.1 mol/L). Weigh 0.4 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water and dilute to 100 mL. Mix well.
4.2.8 Maleic acid buffer (50 mmol/L). Weigh 11.6 g of maleic acid and dissolve it in
1600 mL of water. Adjust to pH=6.0 with 4 mol/L sodium hydroxide solution. Add
another 0.6 g of calcium chloride dihydrate. Add water to dilute to 2 L. Store at 4°C
protected from light. The storage period does not exceed 1 month.
4.2.9 Trishydroxymethylaminomethane (Tris) solution (0.75 mol/L). Weigh 90.8 g of
Tris solid and dissolve it in about 800 mL of water. Add water to dilute to 1 L.
4.2.10 Acetic acid solution (2 mol/L). Measure 115 mL of glacial acetic acid. Add water
to dilute to 1 L.
4.2.11 Mixed enzyme solution. Take 0.5 g of pancreatic α-amylase and 0.05 g of
amyloglucosidase. Use 50 mL of 50 mmol/L maleic acid buffer to prepare a solution
containing 400 U of pancreatic α-amylase and 30 U of amyloglucosidase per mL.
Vortex for 5 min. Prepare when needed.
NOTE. If you need to reduce the amylase hydrolysis time, you can prepare a high-concentration
mixed enzyme solution, that is, the concentrations of pancreatic α-amylase and amyloglucosidase
are 800 U/mL and 340 U/mL respectively.
4.2.12 Protease solution. Take 2.5 g of protease. Use 50 mL of 50 mmol/L maleic acid
buffer to prepare a protease solution containing 50 mg per mL. Vortex for 5 min. Prepare
when needed. Store at 4°C before use.
4.2.13 Pickling diatomite. Take 200 g of diatomite in 600 mL of 2 mol/L hydrochloric
acid. Soak overnight. Filter. Wash with water until the filtrate is neutral. Place it in a
muffle furnace at 550℃±5℃ and burn it before use.
4.2.14 Potassium dichromate lotion. Weigh 100 g of potassium dichromate. Dissolve in
200 mL of water. Place the beaker in cold water to cool, then slowly add 1800 mL of
concentrated sulfuric acid and mix. Stir with a glass rod while adding. Protect against
spills.
4.3 Standard product
4.3.1 Diethylene glycol (C4H10O3). CAS No. 111-46-6, purity ≥99%.
4.3.2 D-glucose (C6H12O6). CAS No.50-99-7, purity ≥99%.
4.3.3 Fructose (C18H32O16). CAS No. 470-69-9, purity ≥98%.
4.3.4 D-Maltose monohydrate (C12H22O11 · H2O). CAS No. 69-79-4, purity ≥98%.
4.4 Preparation of standard solution
4.4.1 Diethylene glycol internal standard solution (100 mg/mL). Accurately weigh 10
g (accurate to 0.1 mg) of diethylene glycol. Dilute with water. Transfer to a 100 mL
volumetric flask. Add water and bring volume to the mark. Prepare when needed.
4.4.2 D-glucose standard solution (10 mg/mL). Accurately weigh 1.0 g (accurate to 0.1
mg) of D-glucose. Dissolve in water and transfer to a 100 mL volumetric flask. Add
water to bring volume to the mark. Prepare when needed.
4.4.3 D-glucose/diethylene glycol solution standard series working solution. Pipette
0.50 mL, 1.0 mL, 2.0 mL, 4.0 mL and 8.0 mL of 10 mg/mL D-glucose standard solution
into 10 mL volumetric flasks. Then add 0.2 mL of 100 mg/mL diethylene glycol internal
standard solution. Add water to dilute and bring volume to the mark. The D...
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GB 5009.88-2023: National food safety standard - Determination of dietary fiber in foods
GB 5009.88-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards -- Determination of dietary
fiber 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 Terms and definitions... 4
3 Principle... 5
4 Reagents and materials... 5
5 Instruments and equipment... 8
6 Analysis steps... 9
Annex A Definition of enzyme activity units and criteria for determining enzyme
activity... 18
Annex B Chromatogram of dietary fiber... 20
National food safety standards -- Determination of dietary
fiber in food
1 Scope
This Standard specifies the method for determination of dietary fiber in food.
This Standard is applicable to the determination of total dietary fiber, soluble dietary
fiber, and insoluble dietary fiber in plant foods and their products, as well as foods with
added dietary fiber components.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 dietary fiber; DF
Carbohydrate polymers that cannot be digested and absorbed by the human small
intestine and have a degree of polymerization ≥3.
According to its source, dietary fiber is divided into. Carbohydrate Polymers naturally
present in the edible parts of plants, such as cellulose, hemicellulose, pectin, lignin, etc.
in plant cell walls; and Carbohydrate Polymers that are isolated, extracted or synthesized
from food raw materials using physical, enzymatic or chemical means and have been
proven by scientific evidence to have beneficial physiological effects.
2.2 soluble dietary fiber; SDF
The portion of dietary fiber that is soluble in water, including indigestible
oligosaccharides and some polysaccharides.
During the detection process, based on whether it can be precipitated by 78% ethanol,
it is divided into precipitable soluble dietary fiber (SDFP) and non-precipitable soluble
dietary fiber (SDFS).
2.3 insoluble dietary fiber; IDF
The portion of dietary fiber that is insoluble in water.
2.4 total dietary fiber; TDF
The sum of soluble dietary fiber and insoluble dietary fiber.
3 Principle
The specimen is homogenized and enzymatically hydrolyzed to remove starch and
protein to obtain an indigestible enzymatic hydrolyzate.
The enzymatic hydrolyzate is precipitated with 78% ethanol. Collect the precipitated
part, wash, dry and weigh it, and then measure the mass of DF (including IDF and SDFP)
in the residue. Collect the filtrate portion. After desalting and concentration, liquid
chromatography (internal standard method) is used to determine SDFS. The sum of the
two is TDF.
The enzymatic hydrolyzate is filtered directly and washed with hot water. The filter
residue is collected, washed, dried, and weighed to determine the mass of the IDF
residue. The filtrate is collected and precipitated with 78% ethanol. The sediment is
dried and weighed. The mass of the SDFP residue is determined. The filtrate fraction is
measured as SDFS. The sum of SDFS and SDFP is SDF.
The mass of TDF, IDF and SDFP residues needs to be deducted from the mass of
residual protein, ash and reagent blank to obtain the dietary fiber content of the
corresponding part.
4 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure, and the
water is grade two water specified in GB/T 6682.
4.1 Reagents
4.1.1 95% ethanol (CH3CH2OH).
4.1.2 Acetone (CH3COCH3).
4.1.3 Petroleum ether. boiling range 30℃~60℃.
4.1.4 Sodium hydroxide (NaOH).
4.1.5 Potassium dichromate (K2Cr2O7).
4.1.6 Trishydroxymethylaminomethane (C4H11NO3, Tris).
4.1.7 Maleic acid (C4H4O4).
4.1.8 Concentrated hydrochloric acid (HCl).
4.1.9 Glacial acetic acid (C2H4O2).
4.2.4 Hydrochloric acid solution (2 mol/L). Measure 167 mL of concentrated
hydrochloric acid. Slowly add 500 mL of water. Mix well and add water to dilute to 1
L.
4.2.5 Sodium hydroxide solution (4 mol/L). Weigh 16 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water to dilute to 100 mL and mix well.
4.2.6 Sodium hydroxide solution (1 mol/L). Weigh 4 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water to dilute to 100 mL and mix well.
4.2.7 Sodium hydroxide solution (0.1 mol/L). Weigh 0.4 g of sodium hydroxide. Slowly
add 60 mL of water. After dissolving, add water and dilute to 100 mL. Mix well.
4.2.8 Maleic acid buffer (50 mmol/L). Weigh 11.6 g of maleic acid and dissolve it in
1600 mL of water. Adjust to pH=6.0 with 4 mol/L sodium hydroxide solution. Add
another 0.6 g of calcium chloride dihydrate. Add water to dilute to 2 L. Store at 4°C
protected from light. The storage period does not exceed 1 month.
4.2.9 Trishydroxymethylaminomethane (Tris) solution (0.75 mol/L). Weigh 90.8 g of
Tris solid and dissolve it in about 800 mL of water. Add water to dilute to 1 L.
4.2.10 Acetic acid solution (2 mol/L). Measure 115 mL of glacial acetic acid. Add water
to dilute to 1 L.
4.2.11 Mixed enzyme solution. Take 0.5 g of pancreatic α-amylase and 0.05 g of
amyloglucosidase. Use 50 mL of 50 mmol/L maleic acid buffer to prepare a solution
containing 400 U of pancreatic α-amylase and 30 U of amyloglucosidase per mL.
Vortex for 5 min. Prepare when needed.
NOTE. If you need to reduce the amylase hydrolysis time, you can prepare a high-concentration
mixed enzyme solution, that is, the concentrations of pancreatic α-amylase and amyloglucosidase
are 800 U/mL and 340 U/mL respectively.
4.2.12 Protease solution. Take 2.5 g of protease. Use 50 mL of 50 mmol/L maleic acid
buffer to prepare a protease solution containing 50 mg per mL. Vortex for 5 min. Prepare
when needed. Store at 4°C before use.
4.2.13 Pickling diatomite. Take 200 g of diatomite in 600 mL of 2 mol/L hydrochloric
acid. Soak overnight. Filter. Wash with water until the filtrate is neutral. Place it in a
muffle furnace at 550℃±5℃ and burn it before use.
4.2.14 Potassium dichromate lotion. Weigh 100 g of potassium dichromate. Dissolve in
200 mL of water. Place the beaker in cold water to cool, then slowly add 1800 mL of
concentrated sulfuric acid and mix. Stir with a glass rod while adding. Protect against
spills.
4.3 Standard product
4.3.1 Diethylene glycol (C4H10O3). CAS No. 111-46-6, purity ≥99%.
4.3.2 D-glucose (C6H12O6). CAS No.50-99-7, purity ≥99%.
4.3.3 Fructose (C18H32O16). CAS No. 470-69-9, purity ≥98%.
4.3.4 D-Maltose monohydrate (C12H22O11 · H2O). CAS No. 69-79-4, purity ≥98%.
4.4 Preparation of standard solution
4.4.1 Diethylene glycol internal standard solution (100 mg/mL). Accurately weigh 10
g (accurate to 0.1 mg) of diethylene glycol. Dilute with water. Transfer to a 100 mL
volumetric flask. Add water and bring volume to the mark. Prepare when needed.
4.4.2 D-glucose standard solution (10 mg/mL). Accurately weigh 1.0 g (accurate to 0.1
mg) of D-glucose. Dissolve in water and transfer to a 100 mL volumetric flask. Add
water to bring volume to the mark. Prepare when needed.
4.4.3 D-glucose/diethylene glycol solution standard series working solution. Pipette
0.50 mL, 1.0 mL, 2.0 mL, 4.0 mL and 8.0 mL of 10 mg/mL D-glucose standard solution
into 10 mL volumetric flasks. Then add 0.2 mL of 100 mg/mL diethylene glycol internal
standard solution. Add water to dilute and bring volume to the mark. The D...