GB/T 28717-2012Determination of malonaldehyde in feed - High performance liquid chromatography (English PDF)
饲料中丙二醛的测定 高效液相色谱法
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
September 3, 2012
Implementation date
February 1, 2013
Scope
GB/T 28717-2012 is the English-translated version of 饲料中丙二醛的测定 高效液相色谱法.
GB/T 28717-2012 is the Chinese standard "Determination of malonaldehyde in feed -- High performance liquid chromatography". Its scope clause reads: This Standard specifies the high-performance liquid chromatography detection method for malondialdehyde in feed.
This Standard is applicable to the determination of malondialdehyde in compound feed, concentrated feed and feed raw materials. The detection limit of this Standard method is 0.015mg/kg, and the quantification limit is 0.05mg/kg.
Its clauses include principle; reagents; apparatus; preparation of specimen; analysis procedure; calculation and presentation of results; repeatability. It was issued by the General Administration of Quality Supervision, Inspection and Quarantine on SEPTEMBER 03, 2012 and took effect on FEBRUARY 01, 2013.
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Document preview — GB/T 28717-2012
National Standard of the People's Republic of China
- ICS
- 65.120
- Classification
- B 46
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- Foreword3
- 1 Scope4
- 2 Normative References4
- 3 Principle4
- 4 Reagents4
- 5 Apparatus5
- 6 Preparation of Specimen6
- 7 Analysis Procedure6
- 8 Calculation and Presentation of Results7
- 9 Repeatability7
- Appendix A (Informative) Chromatogram of Malondialdehyde and Thiobarbituric Acid Complex8
1 Scope
This Standard specifies the high-performance liquid chromatography detection method for malondialdehyde in feed.
This Standard is applicable to the determination of malondialdehyde in compound feed, concentrated feed and feed raw materials.
The detection limit of this Standard method is 0.015mg/kg, and the quantification limit is 0.05mg/kg.
2 Normative References
The following documents are essential to the application of this Document. For the dated documents, only the versions with the dates indicated are applicable to this Document; for the undated documents, only the latest version (including all the amendments) is applicable to this Document.
GB/T 6682 Water for analytical laboratory use - Specification and test methods
GB/T 14699.1 Feeding Stuffs - Sampling
3 Principle
After malondialdehyde in the sample is extracted by trichloroacetic acid, react with thiobarbituric acid (TBA) to form a pink complex. The supernatant is collected by centrifugation; and detect by high-performance liquid chromatography and fluorescence detector. Use external standard method to calculate the content of malondialdehyde.
4 Reagents
Unless otherwise specified, only reagents confirmed to be of analytical grade and Class-I water specified in GB/T 6682 are used in the analysis.
6 Preparation of Specimen
According to the provisions of GB/T 14699.1, take at least 500g of representative feed samples; reduce by at least 100g through quartering method; grind them; pass through a 0.9mm aperture sample sieve; mix them evenly and put them in an airtight container; and store them at low temperature in the dark for later use.
7 Analysis Procedure
7.1 Extraction
Weigh about 2g of specimen (weigh about 1g of feed oil, accurate to 0.001g); put it into a 250mL conical flask; accurately add 50mL of trichloroacetic acid mixture solution (4.4); shake at 180r/min for 30min; take about 20mL of extract into a 50mL centrifuge tube and centrifuge at 5000r/min for 5min.
7.2 Derivatization
Accurately pipette 5mL of the supernatant of the above specimen solution (7.1) and 5mL of malondialdehyde standard series solution (4.7) into 25.0mL colorimetric tubes; add 5.0mL of TBA solution (4.3); mix well; and place in a 90°C water bath to keep warm for 20 min. Take it out; cool it quickly in an ice-water bath; transfer it to a centrifuge tube; centrifuge at 12000r/min for 5 min. Take the supernatant and measure it on a computer; and the malondialdehyde standard series and specimens are derivatized simultaneously.
7.3 Determination
7.3.1 Liquid chromatography reference conditions
7.3.1.1 Chromatographic column: C18 column, 250mm long, 4.6mm inner diameter, 5µm particle size, or equivalent performance.
7.3.1.2 Column temperature: 35°C
7.3.1.3 Mobile phase: Acetonitrile (4.1) + potassium dihydrogen phosphate solution (4.2) = 18 + 82 (V + V); pass through a 0.45µm filter membrane before use.
7.3.1.4 Flow rate: 1.0mL/min.
7.3.1.5 Sample-injection volume: 10µL
7.3.1.6 Excitation wavelength: 525nm
7.3.1.7 Emission wavelength: 560nm […]
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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
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Related Standards
GB/T 14699.1-2005 — Feeding stuffs -- Sampling
GB/T 6682-2008 — Water for analytical laboratory use - Specification and test methods
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