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NY/T 1975-2010 English PDF (NYT1975-2010)

NY/T 1975-2010 English PDF (NYT1975-2010)

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NY/T 1975-2010: Water-soluble fertilizers. Determination of amino-acids content

This standard specifies the automatic amino acid analyzer method and precolumn derivatization-liquid chromatography method for the determination of free amino acids in water-soluble fertilizers. This standard applies to the determination of free amino acid content in liquid or solid water-soluble fertilizers.
NY/T 1975-2010
NY
AGRICULTURE INDUSTRY STANDARD
OF THE PEOPLE REPUBLIC OF CHINA
ICS 65.080
G 20
Water-soluble fertilizers - Determination of amino-
acids contents
ISSUED ON: DECEMBER 23, 2010
IMPLEMENTED ON: FEBRUARY 01, 2011
Issued by: The Ministry of Agriculture of PRC
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Determination of free amino acid content - Amino acid automatic analyzer method (arbitration method) ... 4
4 Determination of free amino acid content - Pre-column derivatization-liquid chromatography method ... 7
Water-soluble fertilizers - Determination of amino-
acids contents
1 Scope
This standard specifies the automatic amino acid analyzer method and pre- column derivatization-liquid chromatography method for the determination of free amino acids in water-soluble fertilizers.
This standard applies to the determination of free amino acid content in liquid or solid water-soluble fertilizers.
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) are applicable to this standard.
GB/T 8170 Rules of rounding off for numerical values and expression and
judgement of limiting values
HG/T 2843 Chemical fertilizer products - Standard volumetric, standard, reagent and indicator solutions for chemical analysis
NY/T 887 Density testing of liquid fertilizer
3 Determination of free amino acid content - Amino
acid automatic analyzer method (arbitration method)
3.1 Principle
After precipitating the protein from the specimen by the use of sulfosalicylic acid, the EDTA complexes metal elements are used to release amino acids. The
amino acids are separated by a separation column, to develop color with ninhydrin, to determine the contents of totally 17 amino acids: aspartic acid, threonine, serine, glutamic acid, Proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine. The sum of these amino acids is the content of free amino acids. 3.2 Reagents and materials
The preparation of reagents, water and solutions used in this standard shall comply with the provisions of HG/T 2843 unless otherwise specified for the specifications and preparation methods.
3.2.1 Sodium citrate (Na3C6H5O7 ?€? 2H2O).
3.2.2 Hydrochloric acid.
3.2.3 Hydrochloric acid solution: c(HCl) = 0.06 mol/L.
3.2.4 Sodium hydroxide solution: ?? (NaOH) = 500 g/L.
3.2.5 Sulfosalicylic acid solution: ?? (C6H6O6S ?€? 2H2O) = 50 g/L.
3.2.6 Disodium edetate solution: ?? (EDTA-Na) = 10 g/L.
3.2.7 Sodium citrate buffer: pH 2.2. Weigh 19.6 g of sodium citrate (3.2.1). Dissolve it. Pipette it to a 1000 mL volumetric flask. Add 16.5 mL of hydrochloric acid (3.2.2). Add water to the mark. Mix it uniformly. If necessary, it may use hydrochloric acid (3.2.2) and sodium hydroxide solution (3.2.4) to adjust the pH to 2.2.
3.2.8 Mixed amino acid standard solution, chromatographically pure. It contains total 17 amino acids: aspartic acid, threonine, serine, glutamic acid, Proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine.
3.3 Instruments
3.3.1 Commonly used laboratory instruments.
3.3.2 Automatic amino acid analyzer.
3.3.3 Centrifuge (above 10000 r/min) or 0.45 ??m microporous membrane, filter and syringe.
3.3.4 Steam drying device: Test tube concentrator or other concentration device. 3.4 Analytical procedures
3.4.1 Preparation of specimens
After the solid sample has been reduced several times, take out about 100 g. Grind it quickly to make all pass through a 0.50 mm aperture sieve (if the sample is wet, it may pass a 1.00 mm sieve). Mix it evenly. Put it in a clean, dry container. After multiple shaking of the liquid sample, quickly take about 200 mL and place After precipitating the protein of specimen by sulfosalicylic acid, use the EDTA to make the metal element release the amino acid. The amino acid is chemically derivatized and separated by liquid chromatography to determine the contents of totally 17 amino acids: aspartic acid, threonine, serine, glutamic acid, Proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine. The sum of these amino acids is the content of free amino acids in the fertilizer.
4.2 Reagents and materials
The preparation of reagents, water and solutions used in this standard shall comply with the provisions of HG/T 2843 unless otherwise the specifications and preparation methods are indicated.
4.2.1 Sulfosalicylic acid solution: ?? (C7H6O6S ?€? 2H2O) = 50 g/L.
4.2.2 Disodium edetate solution: ?? (EDTA - Na) = 10 g/L
4.2.3 Mixed amino acid standard solution, chromatographically pure. It contains total 17 amino acids: aspartic acid, threonine, serine, glutamic acid, Proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine.
4.2.4 Reagents required by the selected derivatization method.
4.3 Instruments
4.3.1 Commonly used laboratory instruments.
4.3.2 Centrifuge (above 10000 r/ min) or 0.45 ??m microporous membrane and filter, syringe.
4.3.3 Liquid chromatograph, equipped with UV detector.
4.3.4 The equipment required for the selected derivative method.
4.4 Analytical procedures
4.4.1 Preparation of specimen
After the solid sample has been reduced several times, take about 100 g and quickly grind it to pass through a 0.50 mm aperture sieve (if the sample is wet, it can pass through a 1.00 mm sieve). Mix it evenly. Place it in a clean, dry container. After shaking the liquid sample many times, quickly take about 100 mL and place it in a clean, dry container.
4.4.2 Preparation of sample solution

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