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    72 Fumaric Acid HPLC

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    bohemian84
    This document describes a method for determining the content of fumaric acid in fruit juices using high-performance liquid chromatography (HPLC). Fruit juice samples are diluted and filtered before being injected into an HPLC system using a column and eluent that allows separation and detection of fumaric acid peaks around 14 minutes. Calibration standards are used to generate a calibration curve to quantify fumaric acid content based on peak area. The method has been validated between laboratories with repeatability and reproducibility precision values reported.

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    72 Fumaric Acid HPLC

    Uploaded by

    bohemian84
    314 views3 pages
    This document describes a method for determining the content of fumaric acid in fruit juices using high-performance liquid chromatography (HPLC). Fruit juice samples are diluted and filtered before being injected into an HPLC system using a column and eluent that allows separation and detection of fumaric acid peaks around 14 minutes. Calibration standards are used to generate a calibration curve to quantify fumaric acid content based on peak area. The method has been validated between laboratories with repeatability and reproducibility precision values reported.

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    This document describes a method for determining the content of fumaric acid in fruit juices using high-performance liquid chromatography (HPLC). Fruit juice samples are diluted and filtered before being injected into an HPLC system using a column and eluent that allows separation and detection of fumaric acid peaks around 14 minutes. Calibration standards are used to generate a calibration curve to quantify fumaric acid content based on peak area. The method has been validated between laboratories with repeatability and reproducibility precision values reported.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    314 views3 pages

    72 Fumaric Acid HPLC

    Uploaded by

    bohemian84
    This document describes a method for determining the content of fumaric acid in fruit juices using high-performance liquid chromatography (HPLC). Fruit juice samples are diluted and filtered before being injected into an HPLC system using a column and eluent that allows separation and detection of fumaric acid peaks around 14 minutes. Calibration standards are used to generate a calibration curve to quantify fumaric acid content based on peak area. The method has been validated between laboratories with repeatability and reproducibility precision values reported.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
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    This document describes the procedure for analyzing fumaric acid content in fruit juices using high performance liquid chromatography (HPLC).

    The content of fumaric acid in fruit juices is determined by high performance liquid chromatography (HPLC).

    Fruit juices are diluted and filtered, then used for HPLC analysis. A standard fumaric acid solution is also prepared.

    International Federation of Fruit Juice Producers IFU

    Fumaric acid
    (HPLC)

    IFU Analyses Nr. 72 (1998) Page 1 of 3

    Principle The content of fumaric acid in fruit juices is determined by high performance liquid chromatography (HPLC).

    WORKING INSTRUCTIONS

    1. 1.1 1.2. 1.3.

    Reagents Double-distilled water or equivalent, suitable for HPLC Solution of sulfuric acid, c (H2SO4) = 0.005 mol/l Fumaric acid, analytical reagent grade

    2. 2.1.

    Apparatus High performance liquid chromatograph with UV-detector and column oven capable of maintaining 40 C HPLC separation column Aminex HPX - 87H (300 x 7.8 mm with precolumn (e.g. micro-guard cartridge Cation H+, 30 x 4.6 mm) Membrane filters (0.45 m or less), suitable for HPLC

    2.2.

    2.3.

    3.

    Determination

    3.1.

    Preparation of test samples

    Fruit juices are diluted 1:20 (concentrates 1 : 100) with water (1.1.) and after membrane filtration (2.3) are used for the HPLC analysis. 3.2. Preparation of standard solution of fumaric acid

    A known weight of fumaric acid (1.3) is dissolved in water (1.1) to give a standard solution of known concentration (fumaric acid) approximately = 100 mg/l.

    International Federation of Fruit Juice Producers IFU

    Fumaric acid
    (HPLC)

    IFU Analyses Nr. 72 (1998) Page 2 of 3

    3.3.

    Preparation of calibration solutions

    The standard solution (3.2) is diluted with water (1.1) to produce calibration standards with (fumaric acid) in the range of 1 mg/l to 10 mg/l. 3.4. Eluent: Column: Conditions for high performance liquid chromatography Sulfuric acid solution , c(H2SO4) = 0.005 mol/l (1.2) Aminex HPX - 87H (300 x 7.8 mm) with precolumn (e.g. micro-guard cartridge Cation H+, 30 x 4.6 mm) 40C Approximately 0.6 ml/min

    Column temperature: Flow:

    Note: To avoid build-up of high pressure, the flow should be increased slowly from 0.2 ml/min to 0.6 ml/min during equilibration of the column with eluent (1.2). Detection Wavelength: Injection volume: Retention time for fumaric acid: Run duration: 210 nm Constant volume of approximately 15 l

    approximately 14 min. approximately 20 min.

    4.

    Calculation and Expression of Results

    The content of fumaric acid is calculated from the peak area of at least three sample injections, using the determined relationship between peak area and fumaric acid content of the calibration solutions (calibration curve or empirically determined mathematical relationship). Take the dilution factor into account in the calculation. The content of fumaric acid is expressed in mg/l to one place of decimals.

    International Federation of Fruit Juice Producers IFU

    Fumaric acid
    (HPLC)

    IFU Analyses Nr. 72 (1998) Page 3 of 3

    5.

    Precision

    Year of collaborative study1996 Number of laboratories 13 Number of samples 3

    Sour cherry nectar Sour cherry nectar Apple juice

    _ x (mg/l) 14.5 3.4 6.8

    sr 0.172 0.165 0.225

    r 0.5 0.5 0.6

    sR 0.674 0.801 0.682

    R 1.9 2.2 1.9

    Repeatability (r): Reproducibility (R):

    sr = 0.190 sR = 0.725

    r = 0.5 mg/l R = 2.0 mg/l

    6.

    Remarks

    The fumaric acid peak is often a rider peak on a matrix peak. In such a case, this should be allowed for when measuring the peak area.

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