GB 5009.210-2023 PDF English
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| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5009.210-2023 | English | 335 |
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National food safety standard - Determination of pantothenic acid in foods
| Valid |
| GB 5009.210-2016 | English | 135 |
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Determination of pantothenic acid in foods
| Obsolete |
| GB/T 5009.210-2008 | English | 439 |
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Determination of pantothenic acid in foods
| Obsolete |
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GB 5009.210-2023: National food safety standard - Determination of pantothenic acid 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/GB5009.210-2023
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National food safety standards -- Determination of
pantothenic acid 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 Principle... 4
3 Reagents and materials... 4
4 Instruments and equipment... 5
5 Analysis steps... 6
6 Expression of analysis results... 7
7 Precision... 8
8 Other... 8
9 Principle... 8
10 Reagents and materials... 8
11 Instruments and equipment... 11
12 Analysis steps... 11
13 Expression of analysis results... 14
14 Precision... 15
15 Other... 15
16 Principle... 15
17 Reagents and materials... 15
18 Instruments and equipment... 18
19 Preparation and preservation of bacterial strains... 18
20 Analysis steps... 19
21 Expression of analysis results... 23
22 Precision... 26
23 Other... 26
Annex A Chromatogram of pantothenic acid standard solution... 27
Annex B Preparation of culture medium (liquid)... 29
Foreword
This Standard replaces GB 5009.210-2016 "Determination of pantothenic acid in
foods".
Compared with GB 5009.210-2016, the main changes in this Standard are as follows.
- Add liquid chromatography-tandem mass spectrometry;
- Modify the pretreatment method for microbial method specimens. Add the
determination method for microplates.
National food safety standards -- Determination of
pantothenic acid in food
1 Scope
This Standard specifies the method for the determination of pantothenic acid in food.
Method One of this Standard is applicable to the determination of pantothenic acid in
infant formula foods (except infant formula foods for special medical purposes), infant
supplementary foods, dairy products, beverages, and nutritional supplements.
Method Two and Method Three of this Standard are applicable to the determination of
pantothenic acid in food.
Method One -- Liquid chromatography
2 Principle
After the specimen is extracted with hot water, it is separated by a C18 reversed-phase
chromatography column, qualitatively determined by retention time, and quantitatively
by the external standard method.
3 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure, and the
water is grade one water specified in GB/T 6682.
3.1 Reagents
3.1.1 Hydrochloric acid (HCl).
3.1.2 Phosphoric acid (H3PO4).
3.1.3 Potassium dihydrogen phosphate (KH2PO4).
3.1.4 Zinc sulfate heptahydrate (ZnSO4 · 7H2O).
3.1.5 Acetonitrile (CH3CN). chromatographically pure.
3.1.6 α-amylase. enzyme activity ≥1.5 U/mg.
3.2 Preparation of reagents
3.2.1 Hydrochloric acid (0.1 mol/L). Pipette 8.3 mL of hydrochloric acid into 800 mL
of water. Add water to dilute to 1000 mL. Mix well.
3.2.3 Zinc sulfate solution (0.5 mol/L). Weigh 14.4 g of zinc sulfate heptahydrate. Add
water to dissolve and dilute to 100 mL.
3.2.4 Potassium dihydrogen phosphate solution (0.02 mol/L). Weigh 2.722 g of
potassium dihydrogen phosphate. Add 500 mL water to dissolve. Use phosphoric acid
to adjust pH to 3.0±0.1.
3.3 Standard product
D-calcium pantothenate standard product (C18H32CaN2O10, CAS number. 137-08-6).
purity ≥99%, or standard product certified by the country and awarded a reference
material certificate.
3.4 Preparation of standard solution
3.4.1 Pantothenic acid standard stock solution (500 μg/mL). Accurately weigh 136 mg
of D-calcium pantothenate standard (accurate to 0.1 mg). Add water to dissolve and
transfer to a 250 mL volumetric flask. Dilute to volume. Mix well. The standard stock
solution shall be stored in the dark at -18°C and below. It can be kept for 3 months.
4 Instruments and equipment
4.1 Balance. resolution is 0.1 mg.
4.2 Constant temperature oscillation water bath. oscillation frequency is 100 r/min±20 r/min.
4.3 Ultrasonic oscillator.
4.4 pH meter. accuracy is ±0.01.
4.5 Centrifuge. rotation speed ≥8000 r/min.
4.6 0.45 μm filter.
4.7 High performance liquid chromatograph. with UV detector or diode array detector.
5 Analysis steps
5.1 Specimen preparation
The solid specimen is crushed and mixed evenly. Carbonated drinks require ultrasonic
removal of carbon dioxide, and other liquid specimens need to be shaken well.
5.2 Specimen extraction
5.2.1 Beverages, nutrient supplements
Accurately weigh or measure an appropriate amount of specimen to the nearest 0.001
g. Generally, the solid specimen is 0.2 g~2 g. The liquid specimen is 10 g~20 g. Place
in a 50 mL Erlenmeyer flask. Add about 30 mL of 40℃~50℃ warm water and conduct
ultrasonic extraction for 20 min.
5.2.2 Infant formula food (except infant formula food for special medical
purposes), infant supplementary food, dairy products
Accurately weigh an appropriate amount of specimen to the nearest 0.001 g. Generally,
the solid specimen is about 5 g.
5.3 Reference chromatographic conditions for liquid phase
The reference chromatographic conditions for the liquid phase are as follows.
5.4 Determination
5.4.1 Determination of standard curve
Carry out chromatographic analysis of the pantothenic acid standard working solution
in sequence (see Figure A.1 in Annex A for the standard chromatogram). Draw a
standard curve with the concentration of the standard working solution as the abscissa
and the peak area as the ordinate.
5.4.2 Determination of specimen solution
The specimen measurement liquid is subjected to chromatographic analysis. Find the
corresponding pantothenic acid concentration from the standard curve based on the
sample peak area.
6 Expression of analysis results
The content of pantothenic acid in the specimen is calculated according to formula (1).
7 Precision
The absolute difference between two independent determination results obtained under
repeatability conditions shall not exceed 10% of the arithmetic mean.
8 Other
8.1 For solid specimens, when the sampling volume is 5 g and the volume is adjusted
to 50 mL, the detection limit is 0.025 mg/100 g and the quantitation limit is 0.08 mg/100 g.
8.2 For liquid specimens, when the sampling volume is 20 g and the volume is adjusted
to 50 mL, the detection limit is 0.0063 mg/100g and the quantification limit is 0.02 mg/100g.
9 Principle
After enzymatic hydrolysis, extraction, centrifugation, and filtration, the specimen is
separated by a C18 reversed-phase chromatography column. Use tandem mass
spectrometry multiple ion reaction monitoring method to test.
10 Reagents and materials
Unless otherwise stated, the reagents used in this method are analytically pure, and the
water is grade one water specified in GB/T 6682.
10.1 Reagents
10.1.1 Acetonitrile (CH3CN). chromatographically pure.
10.1.2 Formic acid (HCOOH). chromatographically pure.
10.1.3 Hydrochloric acid (HCl).
10.1.4 Glacial acetic acid (C2H4O2).
10.1.5 Zinc acetate [Zn (CH3COO)2].
10.1.6 Potassium ferrocyanide [K4Fe (CN)6].
10.1.7 Trishydroxymethylaminomethane (C4H11NO3).
10.2 Preparation of reagents
10.2.1 Formic acid solution (0.1%). Pipette 1 mL of formic acid. Use water to dilute to 1000 mL. Mix well.
10.2.2 Zinc acetate solution (300 g/L). Weigh 30.0 g of zinc acetate. Use water to
dissolve and dilute to 100 mL.
10.2.3 Potassium ferrocyanide solution (150 g/L). Weigh 15.0 g of potassium
ferrocyanide. Use water to dissolve and dilute to 100 mL.
10.2.5 Sodium bicarbonate solution (0.1 mol/L). Weigh 0.84 g of sodium bicarbonate.
Add water to dissolve and dilute to 100 mL. Mix well.
10.2.9 Resin activation. Weigh 100 g of anion exchange resin into a 2 L Erlenmeyer
flask. Add 1 L of 1 mol/L hydrochloric acid solution and shake thoroughly on an
oscillator for 10 min. Filter through a Buchner funnel lined with filter paper. Transfer
the anion exchange resin back to the Erlenmeyer flask. Then add 1 L of 1 mol/L
hydrochloric acid solution, shake repeatedly for 10 min, and filter. Add 1 L of water to
the anion exchange resin and shake for 10 min. filter. Repeat washing with water 10
times. Transfer to the Erlenmeyer flask.
10.3 Standard product
10.3.1 D-calcium pantothenate standard (C18H32CaN2O10, CAS No.. 137-08-6). purity
≥99%, or a standard certified by the country and awarded a reference material certificate.
10.3.2 13C6, 15N2-Calcium Pantothenate (13C6C12H32Ca15N2O10, CAS No.. 356786-94-
2). Purity ≥97%.
10.4 Preparation of standard solution
10.4.1 Pantothenic acid standard stock solution (500 μg/mL). Accurately weigh 136 mg
of D-calcium pantothenate standard (accurate to 0.1 mg). Add water to dissolve and
transfer to a 250 mL volumetric flask. Dilute to volume. Mix well. Standard stock
solutions shall be stored in the dark at -18°C and below.
11 Instruments and equipment
11.1 Liquid chromatography-tandem mass spectrometer. equipped with electrospray
ionization source (ESI).
11.2 Analytical balance. resolution is 0.1 mg and 0.01 g, respectively.
11.3 pH meter. accuracy is ±0.01.
11.6 Pressure steam sterilizer. 121℃.
11.8 Vortex mixer.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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