GB 5009.88-2023 PDF English
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.88-2023 | English | 230 |
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National food safety standard - Determination of dietary fiber in foods
| Valid |
| GB 5009.88-2014 | English | 85 |
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National Food Safety Standard -- Determination of dietary fiber in food
| Obsolete |
| GB/T 5009.88-2008 | English | 439 |
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Determination of dietary fiber in foods
| Obsolete |
| GB/T 5009.88-2003 | English | 199 |
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Determination of insoluble dietary fiber in foods
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GB 5009.88-2023: National food safety standard - Determination of dietary fiber in foods---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GB5009.88-2023
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
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; & 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.
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).
4.1.10 Concentrated sulfuric acid (H2SO4).
4.1.11 Calcium chloride dihydrate (CaCl2 · 2H2O).
4.1.16 Diatomaceous earth. CAS No. 68855-54-9.
4.1.17 Weakly basic acrylic anion exchange resin (OH-). cross-linked acrylic gel whose
functional group is tertiary amine group; average particle size is 0.50 mm~0.75 mm;
ion exchange capacity (based on OH-) ≥1.6 mmol/mL.
4.1.18 Strongly acidic hydrogen ion exchange resin (H+). styrene-divinylbenzene
copolymer whose functional group is sulfonic acid (SO3-); average particle size is 0.82
mm~1.00 mm; ion exchange capacity (calculated as H+) ≥1.6 mmol/mL.
4.2 Preparation of reagents
4.2.1 Ethanol solution (78%). Take 821 mL of 95% ethanol. Dilute with water and bring
to 1 L. Mix well.
4.2.2 Ethanol solution (85%). Take 895 mL of 95% ethanol. Dilute with water and bring
to 1 L. 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.
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.
4.3 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.
4.4.4 Qualitative standard solution. Weigh about 0.10 g of fructose and 0.10 g of D-
maltose monohydrate. Dissolve in water. Transfer to a 50 mL volumetric flask. Add 1
mL of 100 mg/mL diethylene glycol internal standard solution. Add water to bring
volume to the mark. Prepare when needed.
5 Instruments and equipment
5.1 Analytical balance. division is 0.1 mg and 1 mg.
5.2 Dietary fiber measuring device
5.2.1 Vacuum filtration device. Vacuum pump or aspirator with regulating device. 1 L
suction filter bottle with a suction filter port on the side wall and a rubber stopper
matching the suction filter bottle, used for suction filtration of enzymatic hydrolyzate.
5.2.2 Constant temperature oscillating water bath. with automatic timer. The
temperature control range is room temperature +5℃~100℃. The temperature
fluctuation is ±1℃.
5.2.3 Tall beaker without diversion port. 400 mL or 600 mL.
5.2.4 Crucible. with rough sintered glass plate. Pore diameter is 40 μm~60 μm. The
cleaned crucible is ashed in a muffle furnace at 550°C±25°C for 6 h. The furnace
temperature drops below 130℃ and is taken out. Soak in potassium dichromate
washing solution at room temperature for 2 h. Rinse well with tap water and water
respectively. Finally, rinse with 15 mL of acetone and air dry. Before use, add about 1.0
g of diatomaceous earth and bake at 130℃±3℃ until constant weight. Take out the
crucible. Cool in a desiccator for approximately 1 h. Weigh and record the mass of the
crucible (containing diatomite, mG), accurate to 0.1 mg.
5.3 High performance liquid chromatography (HPLC). equipped with a differential
refractive index detector and an 80°C column oven.
5.4 Oven. 40℃±1℃, 105℃±2℃, 130℃±3℃.
5.5 Muffle furnace. 550℃±25℃.
5.6 Rotary evaporator.
5.7 Desiccant. Silica or equivalent desiccant.
5.8 pH meter. with temperature compensation function, accuracy of ±0.1.Calibrate
with pH 4.0, 7.0 and 10.0 standard buffers before use.
5.9 Aqueous phase microporous filter membrane. pore size is 0.45 μm.
5.10 Homogenizer or grinder.
6 Analysis steps
6.1 Preparation of specimen
All specimens need to be homogenized. If necessary, dehydration, defatting or
desugarization can be carried out depending on the moisture, fat and sugar content.
6.1.1 Homogenization
The solid specimen shall be ground, pulverized, and mixed before use. Semi-solid and
liquid specimens shall be homogenized and mixed before use.
6.1.2 Dehydration
For solid, semi-solid and liquid specimens containing floating matter with moisture
content ≥70% and difficult to mix, weigh an appropriate amount of specimen (mC, not
less than 50 g) and dry it in an oven at 105°C±2°C for 4 h. Transfer the specimen to a
desiccator. Weigh (mD) after the specimen temperature drops to room temperature.
According to the mass of the specimen before and after drying, calculate the specimen
mass change factor (f). After drying, the specimen is homogenized and placed in a
desiccator for later use.
6.2 Specimen weighing
Accurately weigh 2 specimens (m) to be tested, accurate to 0.1 mg. The mass difference
between the two specimens is ≤0.005 g. Generally, 0.25 g~3 g of solid specimen is
weighed. Weigh 1.0 g~5.0 g of liquid specimen.
6.3 Enzymatic hydrolysis
Transfer the specimen to a 400 mL~600 mL tall beaker. Add 35 mL of 50 mmol/L
maleic acid buffer. Stir magnetically until the specimen is completely dispersed in the
buffer. Prepare 2 blank samples simultaneously and operate simultaneously.
6.3.1 Enzymatic hydrolysis of amylase
6.3.2 Enzymatic hydrolysis of protease
Add 100 μL of protease solution to each beaker (for animal foods, add 500 μL of
protease solution). Cover with aluminum foil. Place in a 60℃±1℃ water bath and
continue shaking for 30 min.
6.3.3 Addition of internal standard solution
For example, when measuring SDFS, add 2 mL of 100 mg/mL diethylene glycol
internal standard solution to the enzymatic solution.
6.4 Determination of total dietary fiber (TDF)
6.4.1 Precipitation
To the specimen enzymatic hydrolyzate, add 4 times the volume of 95% ethanol that
has been preheated to 60℃±1℃. Remove the beaker. Cover with aluminum foil.
Precipitate at room temperature for 1 h~2 h.
6.4.3 Residue determination
6.4.3.1 Residue mass. The residue is washed twice with 15 mL of 95% ethanol and
twice with 15 mL of acetone. Remove the washing liquid by suction filtration. Place
the crucible and the residue in an oven to dry overnight at 105°C ±2°C. Transfer the
crucible to a desiccator and cool for 1 h.
6.4.4 Determination of SDFS
6.4.4.4 Chromatographic reference conditions
The chromatographic reference conditions are as follows.
6.5 Determination of insoluble dietary fiber (IDF)
6.5.1 Weigh the sample according to 6.2.Carry out enzymatic hydrolysis according to 6.3.
6.6 Determination of soluble dietary fiber (SDF)
6.6.1 Weigh the sample according to 6.2.Carry out enzymatic hydrolysis according to 6.3.
6.6.2 Suction and filter according to 6.5.2.
6.6.3 Collect the filtrate. Collect the filtrate into another pre-weighed 600 mL tall beaker.
Weigh the total mass of "beaker + filtrate". Deduct the mass of the beaker. Estimate the
filtrate volume.
6.7 Expression of analysis results
6.7.1 The mass change factor during specimen preparation is calculated according to
formula (1).
6.7.2 The mass of the residue in the crucible is calculated according to formula (2).
6.7.5 The content of SDFS is calculated according to formula (5).
6.7.6 Expression and conversion relationship of test results
For foods without added dietary fiber components, such as plant foods and their
products, the test results of TDF and SDF do not need to include the SDFS part. TDF
is expressed as TDF (enzyme gravimetric method). The result is equivalent to
TDF=IDF+SDFP. SDF is expressed as SDF (enzyme gravimetric method). The result
6.8 Precision
The absolute difference between two independent determination results obtained under
repeatability conditions shall not exceed 20% of the arithmetic mean.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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