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    Sop For Sodium Selenite Spec Method

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    Ganesh Kashinath
    This document provides the procedure for estimating selenium content in sodium selenite samples using spectrophotometry. It involves reducing selenite to selenium using diaminobenzidine (DAB) reagent, which produces a colored complex. The absorbance of the complex is measured at 420nm and the selenium concentration is calculated based on a standard curve. Key steps include preparing reagents, reducing the selenite sample with DAB, adjusting the pH, extracting the complex into toluene, and measuring absorbance to determine selenium content against a sodium selenite standard curve.

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    Sop For Sodium Selenite Spec Method

    Uploaded by

    Ganesh Kashinath
    248 views3 pages
    This document provides the procedure for estimating selenium content in sodium selenite samples using spectrophotometry. It involves reducing selenite to selenium using diaminobenzidine (DAB) reagent, which produces a colored complex. The absorbance of the complex is measured at 420nm and the selenium concentration is calculated based on a standard curve. Key steps include preparing reagents, reducing the selenite sample with DAB, adjusting the pH, extracting the complex into toluene, and measuring absorbance to determine selenium content against a sodium selenite standard curve.

    Original Description:

    Spec Method for sodium selenite

    Original Title

    Sop for Sodium Selenite Spec Method

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    This document provides the procedure for estimating selenium content in sodium selenite samples using spectrophotometry. It involves reducing selenite to selenium using diaminobenzidine (DAB) reagent, which produces a colored complex. The absorbance of the complex is measured at 420nm and the selenium concentration is calculated based on a standard curve. Key steps include preparing reagents, reducing the selenite sample with DAB, adjusting the pH, extracting the complex into toluene, and measuring absorbance to determine selenium content against a sodium selenite standard curve.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    248 views3 pages

    Sop For Sodium Selenite Spec Method

    Uploaded by

    Ganesh Kashinath
    This document provides the procedure for estimating selenium content in sodium selenite samples using spectrophotometry. It involves reducing selenite to selenium using diaminobenzidine (DAB) reagent, which produces a colored complex. The absorbance of the complex is measured at 420nm and the selenium concentration is calculated based on a standard curve. Key steps include preparing reagents, reducing the selenite sample with DAB, adjusting the pH, extracting the complex into toluene, and measuring absorbance to determine selenium content against a sodium selenite standard curve.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 3

    SI No.

    CONTENTS

    1 Scope of the Document

    2 Principle and Reagent preparation

    3 Selenite reduction

    4 Procedure

    5 Calculation

    6 Reference
    1. The scope of the document: This documentation is applicable for the estimation of selenium in sodium Selenite
    samples.

    2. Principle: The principle is based on the reaction between DAB and tetravalent selenium and spectrophotometrically
    estimating the selenium content.

    2.1. Reagents: Glycine buffer – 0.1 M –Take 0.75 g of glycine dissolve in 100 ml water, then set the pH to 2.6 with
    0.1 N HCl
    DAB: 50 mg in 50 ml of water, 6N HCl -Take 52.6 ml of conc HCL makeup to 100 ml with water.
    Sodium Selenite standard – 100 mg of standard in 20 ml of 6 N HCL. Sample weight is 100 mg.

    3. Reduction of Selenite: Reduction is not applicable; the sample is taken directly to 20 ml of water.

    4. Procedure for Selenite Estimation: From the above take aliquot of 0.01,0.02 and 0.03 and make up the volume
    to 5ml with glycine buffer and add 1 ml DAB reagent, Keep aside for half an hour at room temperature. Set the pH to
    6 using 1 N NaOH and add 10 ml of toluene, vortex until the lower layer is clear, read the OD at 420 nm.

    5. Calculation:
    OD of sample X Conc of standard X Dilution Factor X 100
    in microgram
    OD of Standard
    Weight in mg X 1000

    DF=Dilution Factor

    Numerator 100 = to convert into percentages

    Denominator 100 = weight taken in mg

    1000= convert microgram to mille gram


    Concentration Vs OD
    0.4
    0.35 y = 0.0147x + 0.0695
    R² = 0.9995
    0.3
    Optical Density

    0.25
    0.2
    OD
    0.15
    Linear (OD )
    0.1
    0.05
    0
    0 5 10 15 20 25
    Concentration in microgram

    6. Reference: Spectrophotometric determination of trace elements, Z. Marczenko .

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