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Simultaneous Determination of the Endogenous Free α-Lipoic Acid and Dihydrolipoic Acid in Human Plasma and Erythrocytes by RP-HPLC Coupled with Electrochemical Detector

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Advanced Protocols in Oxidative Stress III

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1208))

Abstract

A highly sensitive, precise, and accurate reversed-phase high performance liquid-chromatography/electrochemical detection method for simultaneous determination of the endogenous free α-lipoic acid and dihydrolipoic acid in biological matrices is presented. The two analytes are extracted from samples with acetonitrile–10 % m-phosphoric acid solution(aqueous) (50:50 v/v). To determine the total lipoic acid, samples are treated with tris(2-carboxyethyl)phosphine solution in phosphate buffer: pH 2.5 with 85 % o-phosphoric acid prior to deproteination. The two analytes are separated on a C18 (150 × 4.6 mm, 5 μm) analytical column using acetonitrile-50 mM phosphate buffer: pH 2.5 with 85 % o-phosphoric acid (35:65 v/v) as the isocratic mobile phase pumped at a flow rate of 2.0 ml/min at the column oven temperature of 35 °C. The column eluents are monitored at a potential of 0.9 V. These analytes are efficiently resolved in <7 min.

Journal published in: Chromatographia, Vol 73, Issue 9, Pages: 929–939, Year of publication: 2011, doi: 10.1007/s10337-011-1997-3.

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Acknowledgement

The authors are thankful to the higher education commission (HEC), Pakistan for funding this study.

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Authors and Affiliations

  1. Department of Pharmacy, University of Peshawar, Peshawar, Pakistan

    Muhammad Imran Khan, Zafar Iqbal & Abad Khan

  2. Department of Pharmacy, Women Institute of Learning, Near Women Medical College, Murree Road, Nawan Shehr, Abbottabad, Pakistan

    Muhammad Imran Khan

  3. Department of Pharmacy, University of Swabi, Swabi, Pakistan

    Abad Khan

Authors
  1. Muhammad Imran Khan
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  2. Zafar Iqbal
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  3. Abad Khan
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Corresponding author

Correspondence to Muhammad Imran Khan .

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Editors and Affiliations

  1. University of Florida, Gainsville, Florida, USA

    Donald Armstrong

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Khan, M.I., Iqbal, Z., Khan, A. (2015). Simultaneous Determination of the Endogenous Free α-Lipoic Acid and Dihydrolipoic Acid in Human Plasma and Erythrocytes by RP-HPLC Coupled with Electrochemical Detector. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress III. Methods in Molecular Biology, vol 1208. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1441-8_25

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  • DOI: https://doi.org/10.1007/978-1-4939-1441-8_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1440-1

  • Online ISBN: 978-1-4939-1441-8

  • eBook Packages: Springer Protocols

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