GB 5413.20-2022 PDF English
US$215.00 · In stock · Download in 9 secondsGB 5413.20-2022: National food safety standard - Determination of Choline in foods, milk and milk products for infants and young children Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedureStatus: Valid GB 5413.20: Evolution and historical versions
| Standard ID | Contents [version] | USD | STEP2 | [PDF] delivery | Name of Chinese Standard | Status |
| GB 5413.20-2022 | English | 215 |
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National food safety standard - Determination of Choline in foods, milk and milk products for infants and young children
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| GB 5413.20-2013 | English | 70 |
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National Food Safety Standard Determination of choline in infant foods and dairy products
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| GB/T 5413.20-1997 | English | 199 |
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Milk powder and formula foods for infant and young children--Determination of choline
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GB 5413.20-2022: National food safety standard - Determination of Choline in foods, milk and milk products for infants and young children
---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/GB5413.20-2022
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
National Food Safety Standard – Determination of Choline in
Foods, Milk and Milk Products for Infants and Young Children
Issued on. JUNE 30, 2022
Implemented on. DECEMBER 30, 2022
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 Apparatus... 6
5 Analysis Procedures... 6
6 Presentation of the Analysis Results... 7
7 Precision... 8
8 Others... 8
9 Principle... 9
10 Reagents and Materials... 9
11 Apparatus... 10
12 Analytical Procedures... 10
13 Presentation of the Analysis Results... 12
14 Precision... 12
15 Others... 13
16 Principle... 13
17 Reagents and Materials... 13
18 Apparatus... 15
19 Analytical Procedures... 15
20 Presentation of Analysis Results... 18
21 Precision... 18
22 Others... 18
Appendix A Ion Chromatogram of Choline Standard Solution... 19
Appendix B Choline Standard Solution Mass Spectrum Scanning Diagram, Standard
Solution and Internal Standard Multi-Reaction Monitoring (MRM) Diagram... 21
Foreword
This Standard replaced GB 5413.20-2013 National Food Safety Standard - Determination of
Choline in Infant Foods and Dairy Products.
Compared with GB 5413.20-2013, the major changes of this Standard are as follows.
--- Delete the second method of Ray's salt spectrophotometry; and add ion chromatography
as the second method;
--- Add liquid chromatography-tandem mass spectrometry as the third method.
National Food Safety Standard – Determination of Choline in
Foods, Milk and Milk Products for Infants and Young Children
1 Scope
This Standard specifies the method for the determination of choline in food, milk and milk
products for infants and young children.
This Standard applies to the determination of choline in food, milk and milk products for infants
and young children.
The First Method – Enzymic Colorimetric Method
2 Principle
The specimen is hydrolyzed by acid; and then reacts with the color developer to form colored
substances after enzymatic action. The alkali content is proportional to the external standard
method. Within a certain concentration range, the color depth is proportional to the choline
content, and the external standard method is used for quantification.
3 Reagents and Materials
Unless otherwise specified, the reagents used in this method are all analytically pure; and the
water is Class-3 water specified in GB/T 6682.
3.1 Reagents
3.1.1 Tris(hydroxymethyl)aminomethane [(CH2OH)3CNH2].
3.1.2 Phenol (C6H5OH).
3.1.3 Concentrated hydrochloric acid (HCl).
3.1.4 Sodium hydroxide (NaOH).
3.1.5 Choline oxidase. ≥10U/mg; store at -20 ℃.
3.1.6 Peroxidase. ≥250U/mg; store at 2℃～8℃.
3.1.7 4-Aminoantipyrine (C11H13N3O).
3.1.8 Phospholipase D. ≥60U/mg, store at -20°C.
3.2 Preparation of reagents
3.3 Standard product
Choline bitartrate standard product (C9H19NO7, relative molecular mass. 253.25, CAS number.
87-67-2). purity ≥ 99%, or the reference material certified by the state and awarded the
reference material certificate.
3.4 Preparation of standard solution
3.5 Materials
4 Apparatus
4.1 Analytical balance. The sensitivities are 0.1 mg and 0.001 g, respectively.
4.3 pH meter. Accuracy 0.01.
4.4 Spectrophotometer.
5 Analysis Procedures
5.1 Sample preparation
5.1.1 Sample pretreatment
Accurately take 20g (accurate to 0.001g) of the mixed liquid specimen into a 100mL conical
flask; add 10mL of 3mol/L hydrochloric acid solution; stopper it and mix well.
Accurately take 5g (accurate to 0.001g) of the mixed semi-solid or solid specimen into a 100mL
conical flask; add 30mL of 1mol/L hydrochloric acid solution; stopper it and mix well.
5.1.3 Filtration
Filter the hydrolyzate with filter paper; if the filtrate is not clear; filter again with a 0.45μm
aqueous filter membrane syringe filter. The filtrate is collected for testing.
5.2 Determination
5.2.1 Drawing of standard curve
Respectively pipette 2.00mL, 4.00mL, 6.00mL, 8.00mL, and 10.00mL of choline standard
working solution (250mg/L) into a 10mL volumetric flask; make constant volume to the mark
with water; mix well; and prepare the standard series working solutions with concentrations of
50.0 mg/L, 100 mg/L, 150 mg/L, 200 mg/L and 250 mg/L, respectively. Prepare 6 colorimetric
tubes; one colorimetric tube is used as reagent blank (A); add 100μL of water; add 100μL of
standard series working solutions to the other 5 colorimetric tubes, respectively; and then add
3.00 mL of color developer, respectively; mix well; the colorimetric tube is placed in a water
bath at 37°C±2°C for 15 min for thermal-insulation reaction.
6 Presentation of the Analysis Results
6.1 Calculation of net absorbance value
Usually formulated reagents shall produce a slight color; and the filtrate is not colorless due to
hydrolysis. In order to remove these interference factors, The respective blank values
(colorimetric tubes A and B) must be subtracted from the total absorbance value.
The net absorbance value of the specimen is calculated according to Formula (1).
6.2 Calculation of choline content
The choline content (by choline hydroxide) in the specimen shall be calculated according to
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
When the solid or semi-solid specimen weighs 5g, the detection limit of the method is
1mg/100g, and the limit of quantification is 3mg/100g.
9 Principle
The specimen is hydrolyzed by acid, purified by solid phase extraction column, then separated
by ion chromatography; detected by conductivity detector, and quantified by external standard
method.
10 Reagents and Materials
Unless otherwise specified, the reagents used in this method are all analytically pure; and the
water is the Class-1 water specified in GB/T 6682.
10.1 Reagents
10.1.1 Concentrated hydrochloric acid (HCl).
10.1.2 Methanesulfonic acid (CH4O3S). chromatographically pure.
10.2 Preparation of reagents
10.2.1 Hydrochloric acid solution (1 mol/L). Pipette 85 mL of concentrated hydrochloric acid
and inject it into about 900 mL of water; and dilute to 1000mL.
10.2.2 Hydrochloric acid solution (1.7 mol/L). Pipette 145 mL of concentrated hydrochloric
acid and inject it into about 800 mL of water; and dilute to 1000mL.
10.3 Standard products
Choline bitartrate standard product (C9H19NO7, relative molecular mass. 253.25, CAS number.
87-67-2). purity ≥ 99%; or a standard substance certified by the state and granted a standard
substance certificate.
10.4 Preparation of standard solutions
10.5 Materials
10.5.1 0.45μm aqueous filter membrane syringe filter.
10.5.2 Purification column. C18 solid phase extraction cartridge 1.0mL or equivalent.
10.5.3 Syringe. 5mL or equivalent.
11 Apparatus
11.1 Ion chromatograph (IC). Equipped with a conductive detector.
11.2 Analytical balance. The sensitivity is 0.1mg and 0.001g, respectively.
11.3 Electrothermal constant temperature water bath device. The temperature can be controlled at 70℃±2℃.
11.4 Vortex mixer.
12 Analytical Procedures
12.1 Sample preparation
12.1.1 Specimen extraction
12.1.2 Specimen purification
C18 solid phase extraction cartridge (1.0mL) is passed through successively with 10mL of
methanol and 15mL of water before use; and stand for activation for 30min. Dilute the
extraction solution 50 times (the dilution ratio can be adjusted appropriately according to the
concentration of choline in the specimen, and it shall be no less than 10 times) with water; take
about 15 mL of the diluted solution; pass through a 0.45μm aqueous filter membrane and a C18
solid phase extraction cartridge (1.0mL); discard the first 3mL; and collect the later eluate for
testing.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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