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    Aoac976 - 01 Biuret

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    Nguyen Hien Duc Hien

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    Aoac976 - 01 Biuret

    Uploaded by

    Nguyen Hien Duc Hien
    162 views1 page

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    AOAC976_01 BIURET

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    162 views1 page

    Aoac976 - 01 Biuret

    Uploaded by

    Nguyen Hien Duc Hien

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    © All Rights Reserved
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    of 1

    2.4.24 tion, 10 mL CuSO4 solution, and 20 mL buffer solution.

    Remove
    AOAC Official Method 976.01 stirring bar, rinse, dilute to volume, mix thoroughly, and let
    Biuret in Fertilizers stand 10 min. With vacuum, filter ca 50 mL through dry 150 mL
    Atomic Absorption Spectrophotometric Method medium porosity fritted glass funnel into dry flask. Transfer 25 mL
    First Action 1976 aliquots of each filtrate to 250 mL volumetric flasks, acidify with
    Final Action 1980 5 mL 1M HCl, and dilute to volume with H2O. Proceed as in
    965.09 (see 2.6.01), using standard solutions, A(h), to determine
    A. Apparatus and Reagents complexed Cu in solution by AA spectrophotometry after adding
    equivalent amounts of alcohol, KOH solution, buffer solution,
    (a) Atomic absorption spectrophotometer.—Instrument with Cu
    and 1M HCl. Take ≥3 readings of each solution. From mean value
    hollow cathode lamp. of Cu concentration, prepare standard curve relating mg Cu found
    (b) Copper sulfate solution.—Dissolve 15 g CuSO4⋅5H2O in to mg biuret added. Redetermine daily.
    H2O and dilute to 1 L. C. Determination
    (c) Buffer solution.—pH 13.4. Dissolve 24.6 g KOH and 30 g
    (a) In urea.—Accurately weigh test portion containing <10 mg
    KCl in H2O and dilute to 1 L.
    biuret, dissolve in H2O, transfer to 100 mL volumetric flask, add
    (d) Starch solution.—Treat 1 g soluble starch with 10 mL cold
    25 mL alcohol, and proceed as in B, beginning “While stirring with
    H2O, triturate to thin paste, and pour gradually into 150 mL boiling
    magnetic stirrer, . . .”. From Cu found, calculate biuret concentra-
    H2O containing 1 g oxalic acid. Boil until solution clears, cool, and
    tion, using standard curve.
    dilute to 200 mL. Prepare fresh weekly.
    (b) In mixed fertilizers.—Transfer accurately weighed test por-
    (e) Bromocresol purple indicator.—Dissolve 0.1 g bromocresol
    tion containing <40 mg biuret to 250 mL beaker and add 1 mL H2O
    purple in 19 mL 0.1M NaOH and dilute to 250 mL with H2O.
    for each g of test sample (5 g maximum). Warm, add 65 mL alcohol
    (f) Biuret.—See 960.04A(c) (see 2.4.23).
    and 7 drops bromocresol purple, and adjust pH to first blue color
    (g) Biuret standard solution.—0.4 mgmL. Dissolve 0.4000 g
    (pH 6–7) with 20% KOH w/v. Place on hot plate, heat to bp, cool,
    recrystallized biuret in warm H2O, cool, transfer to 1 L flask, and di-
    and, if pH has changed, make final adjustment to first blue. Vac-
    lute to volume.
    uum-filter through alcohol-washed paper pulp pad into 100 mL vol-
    (h) Copper standard solutions.—Dilute aliquots of Cu stock so- umetric flask. (If filtrate is not clear, improper pH adjustment has
    lution, 965.09B(b) (see 2.6.01), with H2O to obtain ≥4 standard so- been made. Add HCl and readjust to pH 6–7.) Wash pad and precipi-
    lutions within range of determination, 1–4 µg Cu/mL final tate with alcohol and dilute to volume with alcohol. Transfer 25 mL
    solution. aliquot to 100 mL volumetric flask, and proceed as in B, beginning
    “While stirring with magnetic stirrer, . . .”. From Cu found, calculate
    B. Preparation of Standard Curve biuret concentration, using standard curve and appropriate dilution
    factors. (Final aliquot can be varied to give Cu concentration be-
    Transfer aliquots of biuret standard solution containing 0, 2, tween 1 and 4 µg/mL.)
    4, 6, 8, 10, and 12 mg biuret to separate 100 mL volumetric
    flasks, dilute to ca 30 mL with H2O, and add 25 mL alcohol to References: JAOAC 59, 22(1976); 62, 153(1979); 63, 222(1980).
    each. While stirring with magnetic stirrer, add 2 mL starch solu- CAS-108-19-0 (biuret)

    © 2000 AOAC INTERNATIONAL

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