GB/T 28717-2012 English PDF
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GB/T 28717-2012: Determination of malonaldehyde in feed -- 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/GBT28717-2012GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA ICS 65.120 B 46 Determination of Malonaldehyde in Feed - High Performance Liquid Chromatography ISSUED ON: SEPTEMBER 03, 2012 IMPLEMENTED ON: FEBRUARY 01, 2013 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 ... 4 5 Apparatus ... 5 6 Preparation of Specimen ... 6 7 Analysis Procedure ... 6 8 Calculation and Presentation of Results ... 7 9 Repeatability ... 7 Appendix A (Informative) Chromatogram of Malondialdehyde and Thiobarbituric Acid Complex ... 8 Determination of Malonaldehyde in Feed - High Performance Liquid Chromatography1 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 – Sampling3 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. ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.
