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    Lutein From Tagetes Erecta: Synonyms

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    Krrlive
    This document provides specifications for lutein extracted from marigold flowers (Tagetes erecta). It establishes an acceptable daily intake of 0-2 mg/kg body weight and defines lutein as a purified extract containing mainly lutein and other carotenoids. The document also specifies assay methods to analyze lutein content and purity, limiting substances like moisture, ash, zeaxanthin, lead and residual solvents.

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    Lutein From Tagetes Erecta: Synonyms

    Uploaded by

    Krrlive
    85 views4 pages
    This document provides specifications for lutein extracted from marigold flowers (Tagetes erecta). It establishes an acceptable daily intake of 0-2 mg/kg body weight and defines lutein as a purified extract containing mainly lutein and other carotenoids. The document also specifies assay methods to analyze lutein content and purity, limiting substances like moisture, ash, zeaxanthin, lead and residual solvents.

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    This document provides specifications for lutein extracted from marigold flowers (Tagetes erecta). It establishes an acceptable daily intake of 0-2 mg/kg body weight and defines lutein as a purified extract containing mainly lutein and other carotenoids. The document also specifies assay methods to analyze lutein content and purity, limiting substances like moisture, ash, zeaxanthin, lead and residual solvents.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    85 views4 pages

    Lutein From Tagetes Erecta: Synonyms

    Uploaded by

    Krrlive
    This document provides specifications for lutein extracted from marigold flowers (Tagetes erecta). It establishes an acceptable daily intake of 0-2 mg/kg body weight and defines lutein as a purified extract containing mainly lutein and other carotenoids. The document also specifies assay methods to analyze lutein content and purity, limiting substances like moisture, ash, zeaxanthin, lead and residual solvents.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
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    of 4

    LUTEIN from TAGETES ERECTA

    New specifications prepared at the 63rd JECFA (2004) and published


    in FNP52 Add 12 (2004). A group ADI of 0 - 2 mg/kg bw for lutein from
    T. erecta and synthetic zeaxanthin was established at the 63rd JECFA
    (2004).

    SYNONYMS Vegetable lutein; vegetable luteol; Bo-Xan (lutein), INS No. 161b(i)

    DEFINITION Lutein from Tagetes erecta L. is a purified extract of xanthophylls


    obtained from marigold oleoresin. The oleoresin is prepared from
    hexane extracts of marigold (Tagetes erecta L) flowers, saponified
    with potassium hydroxide in either methanol or propylene glycol. The
    resulting crystalline material contains lutein, and minor components
    including other carotenoids and waxes.

    Chemical names 3R,3’R,6’R-β,ε-carotene-3,3'-diol; all-trans-lutein; 4’,5’-didehydro-5’,6’-


    dihydro-beta,beta-carotene-3,3’-diol (lutein)

    C.A.S. number 127-40-2 (lutein)

    Chemical formula C40H56O2 (lutein)

    Structural formula

    Formula weight 568.88 (lutein)

    Assay Not less than 80 % total carotenoids, not less than 70 % lutein

    DESCRIPTION A free-flowing, orange-red powder

    FUNCTIONAL USES Colour, nutrient supplement

    CHARACTERISTICS
    IDENTIFICATION

    Solubility (Vol. 4) Insoluble in water, soluble in hexane

    Spectrophotometry A chloroform/ethanol (1:9) solution shows maximum absorbance at ca.


    (Vol. 4) 445 nm
    Melting range (Vol. 4) 177 to 178º

    Test for carotenoids The colour of a solution of the sample in acetone disappears after
    (Vol. 4) successive addition of a 5% solution of sodium nitrite and 0.5 M of
    sulfuric acid.
    PURITY

    Moisture (Vol. 4) Not more than 1.0%

    Ash (Vol. 4) Not more than 1.0%

    Zeaxanthin Not more than 9.0%.


    See description under METHOD OF ASSAY

    Lead (Vol. 4) Not more than 3 mg/kg.


    Determine using an atomic absorption technique appropriate to the
    specified level. The selection of sample size and method of sample
    preparation may be based on the principles of the methods described
    in Volume 4, "Instrumental Methods"

    Hexane (Vol. 4) Not more than 50 mg/kg

    Methanol (Vol. 4) Not more than 10 mg/kg

    Propylene glycol Not more than 1000 mg/kg


    Test as described for Sucrose Esters of Fatty Acids (FNP 52 Add 11 p
    76)

    Waxes Not more than 14.0%


    See description under TESTS.

    TESTS
    PURITY TESTS

    Waxes Determine by gas chromatography using the following conditions:


    Apparatus
    GC equipped with an autosampler, a splitless injection system, flame
    ionization detector (FID), programmable column and detector flow
    rates.
    GC column DB5, 30 m x 0.25 mm ID with a 0.25 µm
    film thickness.
    GC injector temperature: 280°
    FID temperature: 300°
    GC column initialtemperature: 50° (held for 2 min)
    GC oven temperature increase rate: 13°/min
    GC column final temperature:300° (held for 8 min)
    Carrier gas (Helium) flow rate: 1.0 ml/min
    Injection mode: splitless
    Approximate run time: 30 min

    Internal standard pentacosane (C25)


    Calibration standards are prepared through the addition of absolute
    hydrocarbon standards to methylene chloride to provide hydrocarbon
    concentrations of 2.0, 10, 25, 50, 75, and 100 mg/kg.
    Sample Preparation
    Accurately weigh 200 mg of sample into a centrifuge tube and dissolve
    in exactly 20 ml of methylene chloride. Sonication or vortex mixing
    may be required to completely dissolve the product.
    Centrifuge sample at 2500 rpm for 5 min if the sample appears turbid.
    Transfer 40 µl into 2 ml autosampler vial that contains 1.6 ml of
    methylene chloride and 20 µl of (5000 mg/kg) pentacosane for a final
    concentration of 50 mg/kg.
    Sample Analysis
    Autosampler injects a 1.0 µl aliquot of the solution onto the GC
    column.
    Results
    The approximate retention according to GC/FID times of nonacosane
    (C29), triacontane (C30), henitriacontane (C31), C32, triatriacontaine
    (C33), C34, C35, and the internal standard pentacosane (C25) are
    18.6, 19.1, 19.6, 20.0, 20.5, 20.9, 21.4, and 16.3 minutes,
    respectively.

    METHOD OF Determine the total carotenoid content and the content of lutein and
    ASSAY zeaxanthin by HPLC using the following conditions:
    Reagents:
    Hexane (HPLC grade)
    Ethyl acetate (HPLC grade)
    Ethyl alcohol
    Toluene
    Solvent Mixture: (10:6:7:7 hexane:ethanol:acetone:toluene v/v/v/v).
    Standard Solution:
    Weigh accurately about 1g lutein and transfer into 100 ml amber
    volumetric flask and dilute to mark with the Solvent Mixture.
    Apparatus
    UV/vis spectrophotometer
    HPLC system with suitable diode array detector, autosampler, column
    oven, signal processor and degasser.
    Analytical column: 3 µm silica, 4.6 mm x 250 mm.
    Instrument Conditions
    Oven temperature: ambient
    Mobile Phase: 70:30 (v:v) hexane/ethyl acetate (isocratic
    elution)
    Flow Rate: 1.5 ml/min
    Injection: 10 µl
    Detection: performed at 446 nm
    Run Time: approximately 40 min

    Sample Preparation:
    Weigh sample (range 27 to 33 mg) into a glass weighing funnel, wash
    crystals with heat into a 100 ml volumetric flask, dilute to the mark and
    stir for 10 min. Pipette 1 ml from flask into a second 100 ml volumetric
    flask, dilute to the mark with ethanol, mix by inversion for 20 seconds.
    Read samples in a spectrophotometer at 446 nm.

    For HPLC, dry the samples down using nitrogen steam, dissolve
    solids in 70:30 hexane:ethyl acetate, add 0.5 ml to HPLC vials and
    measure at 446 nm.
    Results
    The retention times for lutein and zeaxanthin are approximately 7.7
    and 8.1 min, respectively. The resolution between the HPLC peaks for
    lutein and xeazanthin ranged from 3.06 to 3.09.
    Calculation

    Absorbance at 446 nm x 10000


    Total carotenoids (%) =
    sample mass in g x 2550

    Note: The factors 10000 and 2550 are the dilution factor and extinction
    value for a 1% solution, respectively.

    Lutein (%) = total carotenoids x area % lutein


    Zeaxanthin (%) = total carotenoids x area % zeaxanthin

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