GB/T 18872-2017 PDF English
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| GB/T 18872-2017 | English | 115 |
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Determination of vitamin K3 in feeds - High performance liquid chromatography
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| GB/T 18872-2002 | English | 279 |
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Determination of vitamin K3 in feeds -- High-performance liquid chromatography
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GB/T 18872-2017: Determination of vitamin K3 in feeds - High performance liquid chromatography---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/GBT18872-2017
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 65.120
B 46
Replacing GB/T 18872-2002
Determination of vitamin K3 in feeds - High performance
liquid chromatography
Issued on. NOVEMBER 01, 2017
Implemented on. MAY 01, 2018
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine;
Standardization Administration of the People's Republic of China.
Table of Contents
Foreword... 3
1 Scope... 4
2 Normative references... 4
3 Principle... 4
4 Reagents and materials... 4
5 Instruments and equipment... 5
6 Sampling and specimen preparation... 6
7 Analysis steps... 6
8 Result calculation... 7
9 Repeatability... 8
Annex A (informative) Example of labeled amount of feed sample, weighed sample
amount and dilution ratio of menadione extract... 9
Annex B (informative) Chromatogram of menadione standard solution... 10
Foreword
This Standard was drafted in accordance with the rules given in GB/T 1.1-2009.
This Standard replaces GB/T 18872-2002 “Determination of Vitamin K3 in feeds-High-
performance liquid chromatography”. Compared with GB/T 18872-2002, the main
technical changes in this Standard are as follows.
- Supplemented the concentrate supplement in the scope of application; Added the
varieties of feed additive vitamin K3, that is, this method is also applicable to
vitamin K3 added in the form of nicotinamide bisulfite menadione and
dimethylpyrimidinol menadione sulfite;
- In the scope, it specifies that the content of vitamin K3 is calculated as menadione;
Revised the measurement range above 2mg/kg to the limit of quantification
0.4mg/kg;
- Improved the sample extraction methods from ammonia water hydrolysis to
sodium carbonate solution hydrolysis;
- Added the conversion coefficient between menadione sodium bisulfite,
nicotinamide bisulfite menadione, dimethylpyrimidinol menadione sulfite and
menadione.
This Standard was proposed by and shall be under the jurisdiction of National Technical
Committee on Feed Industry of Standardization Administration of China (SAC/TC 76).
The drafting organizations of this Standard. Institute of Agricultural Quality Standards
and Testing Technology, Chinese Academy of Agricultural Sciences [National Feed
Quality Supervision and Inspection Center (Beijing)], Guangzhou Aibao Agricultural
Feed Co., Ltd., China Animal Husbandry Industry Co., Ltd.
Main drafters of this Standard. Zhao Xiaoyang, Liu Zhiying, Zhu Gaoqun, Wang
Zhongyan, Guo Hongshuang.
Version of standard substituted by this Standard is.
- GB/T 18872-2002.
Determination of vitamin K3 in feeds - High performance
liquid chromatography
1 Scope
This Standard specifies the high performance liquid chromatography for the
determination of vitamin K3 (menadione sodium bisulfite, nicotinamide bisulfite
menadione, dimethylpyrimidinol menadione sulfite, as menadione) in feeds.
This Standard applies to the determination of vitamin K3 in compound feeds,
concentrated feeds, additive premixed feeds and concentrate supplements. The limit of
quantification of the method is 0.4mg/kg.
2 Normative references
The following referenced documents are indispensable for the application of this
document. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
GB/T 603, Chemical reagent - Preparations of reagent solutions for use in test
methods
GB/T 6682, Water for analytical laboratory use - Specification and test methods
GB/T 14699.1, Feeding Stuffs - Sampling
GB/T 20195, Animal feeding stuffs - Preparation of test samples
3 Principle
The specimen is extracted with chloroform and sodium carbonate solution and
converted into free menadione. It is separated by reversed-phase C18 column, detected
by UV detector, and quantified by external standard method.
4 Reagents and materials
Unless otherwise stated, all reagents in this Standard are analytically pure. The test
water is in line with the requirements for grade three water in GB/T 6682.
4.1 Trichloromethane.
4.2 Methanol. chromatographically pure.
4.5 Anhydrous sodium sulfate.
4.6 Diatomaceous earth.
4.7 Diatomaceous earth and anhydrous sodium sulfate mixture. Weigh 3g of
diatomaceous earth and mix with 20g of anhydrous sodium sulfate.
4.8 Menaquinone standard. content is ≥ 96%.
4.10 Menadione standard working solution. Accurately pipette 1.00mL of menadione
standard stock solution (4.9) into a 100mL brown volumetric flask. Use methanol to
dissolve. Dilute to the scale. Mix well. The concentration of the working solution is
about 5μg/mL. Store at -18°C. It is valid for 3 months.
5 Instruments and equipment
5.1 Commonly used laboratory equipment.
5.2 Balance. resolution is 0.001g.
5.3 Balance. resolution is 0.0001g.
5.4 Balance. resolution is 0.00001g.
5.8 High performance liquid chromatograph. with UV tunable wavelength detector (or
diode matrix detector).
6 Sampling and specimen preparation
Take representative feed samples according to GB/T 14699.1.Conduct down-sampling
by quartering.
7 Analysis steps
WARNING -- Because vitamin K3 is sensitive to air and ultraviolet light, and the
chloroform solution used as an extractant has certain toxicity, all operations
should be performed in a fume hood away from light.
7.1 Preparation of specimen solutions
Weigh 0.25g~0.5g of vitamin premix feed (accurate to 0.0001g) or 1g of compound
premix feed or 5g of concentrated feed (accurate to 0.001g), 5g~10g of compound feed
and concentrate supplement (accurate to 0.001g).
7.2 Determination
7.2.1 Reference chromatographic conditions
Chromatographic column. C18 type column; length is 150mm, inner diameter is 4.6mm,
particle size is 5μm, or analytical column with similar performance;
Mobile phase. methanol + water (75+25);
7.2.2 Quantitative determination
Inject the corresponding menadione standard working solution (4.10) and sample
solution (7.1) in turn. Obtain chromatographic peak area response values. Use external
standard method to carry out quantitative determination. See Figure B.1 for the
chromatogram of the standard solution of menadione.
8 Result calculation
The content X of menaquinone in the specimen, expressed as the mass fraction of
menaquinone in the sample, in milligrams per kilogram (mg/kg), is calculated
according to formula (1).
9 Repeatability
For feeds with vitamin K3 content less than 100mg/kg, the difference between the two
independent determination results obtained under repeatability and their arithmetic
mean is not more than 20% of the arithmetic mean of the two determinations.
Annex A
(informative)
Example of labeled amount of feed sample, weighed sample amount and dilution
ratio of menadione extract
The example of the labeled amount, weighed sample amount and the dilution ratio of
the menadione extract of the feed sample are shown in Table A.1.
Annex B
(informative)
Chromatogram of menadione standard solution
The chromatogram of the standard solution of menadione is shown in Figure B.1.
......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
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