GB 5009.89-2023 PDF English
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.89-2023 | English | 260 |
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National food safety standard - Determination of niacin and niacinamide in foods
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| GB 5009.89-2016 | English | 85 |
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National Food Safety Standard -- Determination of Niacin and Nicotinamide in Foods
| Obsolete |
| GB/T 5009.89-2003 | English | 239 |
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Determination of niacin in foods
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GB 5009.89-2023: National food safety standard - Determination of niacin and niacinamide 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/GB5009.89-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard - Determination of Niacin
and Niacinamide in Foods
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.
Table of Contents
Foreword... 3
1 Scope... 4
Method I - High Performance Liquid Chromatography... 4
2 Principle... 4
3 Reagents and Materials... 4
4 Instruments and Equipment... 5
5 Analytical Procedures... 6
6 Expression of Analysis Results... 8
7 Precision... 9
8 Others... 9
Method II - Microbiological Method... 9
9 Principle... 9
10 Reagents and Materials... 9
11 Instruments and Equipment... 11
12 Analytical Procedures... 12
13 Expression of Analysis Results... 17
14 Precision... 18
15 Others... 18
Appendix A Liquid Chromatogram of Niacin and Niacinamide Standard Solution... 19
Appendix B Preparation of Culture Medium... 20
1 Scope
This Standard specifies the methods for the determination of niacin and niacinamide in foods.
In this Standard, Method 1 is applicable to the determination of niacin and niacinamide in
prepared milk powder, special dietary foods (excluding partially hydrolyzed milk protein
formulas, deeply hydrolyzed milk protein formulas or amino acid formulas, and infant formulas
for special medical purposes for amino acid metabolism disorders) and special-purpose
beverages. Method 2 is applicable to the determination of niacin (or niacinamide) in foods.
Method I - High Performance Liquid Chromatography
2 Principle
After pre-treatment, such as. enzymatic hydrolysis and protein precipitation, the sample is
extracted through ultrasonic oscillation in a weakly acidic environment, separated by C18
chromatographic column and detected by a UV detector. In accordance with the retention time
of the chromatographic peak, conduct qualitative determination, and adopt the external standard
method for quantitative determination. Calculate the content of niacin and niacinamide in the
specimen.
3 Reagents and Materials
Unless it is otherwise specified, the reagents used in this Method are all analytically pure, and
the water is Grade-1 water specified in GB/T 6682.
3.1 Reagents
3.1.1 Hydrochloric acid (HCl). guaranteed reagent.
3.1.2 Sodium hydroxide (NaOH). guaranteed reagent.
3.2 Preparation of Reagents
3.2.1 Hydrochloric acid solution (5.0 mol/L). measure-take 415 mL of hydrochloric acid and
add water to reach a constant volume of 1,000 mL.
3.2.2 Hydrochloric acid solution (0.1 mol/L). draw-take 8.3 mL of hydrochloric acid and add
water to reach a constant volume of 1,000 mL.
3.3 Reference Materials
3.3.1 Niacin (C6H5NO2, CAS. 59-67-6). purity 98%, or a standard substance certified by the
state and awarded a reference material certificate.
3.4 Preparation of Standard Solutions
3.4.2 Niacin and niacinamide standard mixed intermediate solution (100.0 g/mL). respectively
and accurately draw-take 10.0 mL of niacin and niacinamide standard stock solution into a 100
mL volumetric flask, add water to reach a constant volume to the scale, and evenly mix it. It
can be stored for 1 month when refrigerated at 2 C ~ 8 C.
3.4.3 Niacin and niacinamide standard mixed working solutions. respectively and accurately
draw-take 1.0 mL, 2.0 mL, 5.0 mL, 10.0 mL and 20.0 mL of the standard mixed intermediate
solution into 100 mL volumetric flasks, add water to reach a constant volume to the scale and
evenly mix them. Thus, standard mixed working solutions with a mass concentration of 1.0
g/mL, 2.0 g/mL, 5.0 g/mL, 10.0 g/mL and 20.0 g/mL are obtained. Prepare them right
before use.
4 Instruments and Equipment
4.1 High performance liquid chromatograph. equipped with UV detector or diode array detector.
4.2 Balance. with a division value of 0.1 mg and 0.01 g, respectively.
4.4 Ultrasonic equipment.
4.5 pH meter. with an accuracy of 0.1.
4.6 Pulverizer.
5 Analytical Procedures
5.1 Sample Pre-treatment
5.1.1 Specimen preparation
Sample pre-treatment. take at least 200 g of representative sample. For lumpy or granular
samples, use a pulverizer to pulverize them; for powdery, pasty or liquid samples, thoroughly
mix them and place them in a closed container.
5.1.2 Starches and starch-containing foods
5.1.3 Starch-free foods
5.2 Reference Conditions of Liquid Chromatography
5.3 Drawing of Standard Curve
In accordance with the chromatographic conditions that have been established, successively
determine the niacin and niacinamide mixed standard series of determination solutions based
on the above recommended chromatographic conditions (see Figure A.1 in Appendix A for the
standard sample chromatogram). Record the chromatographic peak area of each component,
take the mass concentration of the standard determination solutions as the x-coordinate, and the
peak area as the y-coordinate to draw a standard curve.
5.4 Determination of Specimen Solution
Inject the specimen determination solution for determination in accordance with the above
recommended chromatographic conditions.
6 Expression of Analysis Results
6.1 Calculation of Niacin or Niacinamide Content in the Specimen
The content of niacin or niacinamide in the specimen is calculated in accordance with Formula (1).
6.2 Calculation of Total Niacin and Niacinamide Content in the Specimen
The total content (calculated by niacin) of niacin and niacinamide in the specimen is calculated
in accordance with Formula (2).
7 Precision
The absolute difference between the results of two independent determinations obtained under
repeatability conditions shall not exceed 10% of the arithmetic mean.
8 Others
For solid samples. when the sampling size is 5 g, the detection limit of niacin is 30 g/100 g,
and the quantitation limit is 100 g/100 g; the detection limit of niacinamide is 40 g/100 g,
and the quantitation limit is 120 g/100 g.
9 Principle
Niacin and niacinamide are essential nutrients for the growth of Lactiplantibacillus plantarum.
In the niacin determination culture medium, the growth of Lactiplantibacillus plantarum is
correlated with the niacin (or niacinamide) content.
10 Reagents and Materials
Unless it is otherwise specified, the reagents used in this Method are all analytically pure, and
the water is Grade-1 or Grade-2 water specified in GB/T 6682.
10.1 Strain
Lactiplantibacillus plantarum (the former Lactobacillus plantarum) ATCC 8014, or equivalent
strain.
10.2 Culture Media
10.2.1 Lactobacillus agar culture medium. see B.1 in Appendix B.
10.3 Reagents
10.3.1 Absolute ethanol (C2H5OH).
10.3.2 Sulfuric acid (H2SO4). 95% ~ 98%.
10.3.3 Sodium hydroxide (NaOH).
10.3.4 Sodium chloride (NaCl).
10.4 Preparation of Reagents
10.4.1 Ethanol solution (with a volume fraction of 25%). measure-take 250 mL of absolute
ethanol, add water to reach a constant volume of 1,000 mL.
10.4.2 Sulfuric acid solution A (10 mol/L). measure-take 560 mL of sulfuric acid, add it to water
and dilute it to 1,000 mL.
10.4.3 Sulfuric acid solution B (0.5 mol/L). measure-take 50 mL of sulfuric acid A (10 mol/L),
add it to water and dilute it to 1,000 mL.
10.4.4 Sodium hydroxide solution A (10 mol/L). weigh-take 400 g of sodium hydroxide, add
water to dissolve it and dilute to 1,000 mL.
10.5 Reference Materials
10.5.1 Niacin (C6H5NO2, CAS. 59-67-6). purity 98%, or a standard substance certified by the
state and awarded a reference material certificate.
10.5.2 Niacinamide (C6H6N2O, CAS. 98-92-0). purity 98%, or a standard substance certified
by the state and awarded a reference material certificate.
10.6 Preparation of Standard Solutions
10.6.1 Niacin (or niacinamide) standard stock solution (50 g/mL). place niacin (or
niacinamide) reference material into a desiccator containing phosphorus pentoxide and dry it
overnight. In accordance with purity, weigh it, so that the niacin (or niacinamide) content is
50.0 mg (accurate to 0.001 g), use ethanol solution (25%) to dissolve it and transfer it to a 1,000
mL volumetric flask, and reach a constant volume to the scale.
10.6.2 Niacin (or niacinamide) standard intermediate solution (500 ng/mL). draw-take 1.0 mL
of niacin (or niacinamide) standard stock solution (50 g/mL) into a 100 mL volumetric flask,
use ethanol solution (25%) to reach a constant volume to the scale.
11 Instruments and Equipment
Except for the conventional sterilization and culture equipment of microbiology laboratory,
other equipment and materials are as follows.
11.1 Analytical balance. with a division value of 0.1 mg.
11.2 Centrifuge.
11.3 Vortex mixer.
11.4 pH meter. with an accuracy of 0.01.
11.12 Volumetric flask. with a capacity of 100 mL, 200 mL, 250 mL and 500 mL.
11.13 One-mark pipette. 1 mL, 5 mL and 10 mL.
11.14 Graduated pipette. 5 mL (with a scale of 0.1 mL).
11.15 Glass funnel. with a diameter of 100 mm.
11.16 Conical flask. with a capacity of 250 mL.
11.17 Beaker. with a capacity of 100 mL.
11.18 Dispenser. 0 mL ~ 10 mL.
11.19 Micropipette. 1,000 L and 200 L.
11.20 Sterile centrifuge tube. 1.5 mL.
11.21 Syringe filter. with an aperture of 0.22 m.
12 Analytical Procedures
12.1 Preparation of Test Bacterial Suspension
12.2 Specimen Extraction
Lumpy and granular specimens need to be crushed; powdered specimens, such as. milk powder
and rice flour, need to be evenly mixed; fruits and vegetables, meat, eggs, fish and animal offal,
etc., need to be made into chyme; semi-solid foods need to be homogenized and evenly mixed;
liquid specimens shall be shaken and mixed before use.
12.2.4 Dilution. in accordance with the niacin (or niacinamide) content in the specimen, use
water to appropriately dilute the extracting solution, so that the mass concentration of niacin
(or niacinamide) in the specimen extracting solution after dilution is 5 ng/mL ~ 12 ng/mL.
12.3 Specimen Determination
12.3.1 Test tube culture method
12.3.1.2 Specimen series of tubes
In accordance with the sequence listed in Table 2, add water, specimen extracting solution and
culture medium for niacin determination to the culture tube. Prepare 3 tubes for each number
in the Table.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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