HPLC analysis of an ascorbic acid in biological fluids and some foodstuff

  • Alexei A. Dutov МD (clinical pharmacology), physician of the highest qualifying category (clinical laboratory diagnostics), Senior Research Fellow Laboratory of Experimental and Clinical Biochemistry and Immunology, Research Institute of Medical Ecology of the Medical Academy, Chita. e-mail: dutovaa@yandex.ru
  • Denis A. Nikitin head of the laboratory of chemical analysis of the Zabaikalsky State University, Chita, e-mail: nikitinnd@gmail.com
  • Anna M. Akhmetshina post-graduate student Department of Chemistry 1th-year training Zabaikalsky State University, Chita
  • Olga M. Akhmetshina post-graduate student Department of Chemistry 1th-year training Zabaikalsky State University, Chita
  • Anastasia V. Martynova post-graduate student Department of Chemistry 1th-year training Zabaikalsky State University, Chita
  • Anna V. Sverkunova post-graduate student of chemistry 1th-year learning the Zabaikalsky State University, Chita
  • Alena V. Yermolina physician ophthalmologist Regional Hospital № 2, Chita
  • Ekaterina N Fedorova post-graduate student Department of Chemistry, 3th-year training Zabaikalsky State University, Chita
  • Olga N. Konovalova post-graduate student Department of Chemistry, 3th-year training Zabaikalsky State University, Chita
  • Julia L. Lukyanova a physician ordinator Department of Neurology, Medical Academy, Chita
  • Maria N. Mishchenko cand. sci. med., assistant Department of Dentistry, Medical Academy, Chita
  • Anastasia N. Semenova post-graduate student Surgical Dentistry 3th year of study Medical Academy, Chita
Keywords: HPLC, UV and fluorimetric detection, ascorbic acid, plasma, saliva, plaintive liquid (tear), foodstuff.

Abstract

An HPLC method for measuring total ascorbic acid in human plasma, saliva, plaintive liquid (or tear)
and some foodstuff was validated. Biological samples were deproteinized and cleared using acetonitrile and
chloroform. Ascorbic acid oxidized in dehydroascorbic acid by means of iodine solution. The further
derivatization was carried out o-phenylenediamine under the mild conditions (15 min, 400C). The derivatives
were separated by reversed phase (C18) HPLC with UV 365 nm and fluorimetric detection at ex365-em418
nm. We are used high-speed columns (100 × 4.6 mm) Chromolith Performance RP-18e from Merck. The
mobile phase consisted of 20 % acetonitrile and 0.05 % trifluoroacetic acids. Full separation was reached less
than for 2 minutes at speed of a flow of 1400 μl/min. Simplicity, reproducibility and high sensitivity of a
method, allow its use in a clinical practice for an estimation of biochemical changes at eye and stomatologic
diseases and also for measurements of concentration of an ascorbic acid in berries, fruit and juices.

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Published
2019-11-20
How to Cite
Dutov, A. A., Nikitin, D. A., Akhmetshina, A. M., Akhmetshina, O. M., Martynova, A. V., Sverkunova, A. V., Yermolina, A. V., Fedorova, E. N., Konovalova, O. N., Lukyanova, J. L., Mishchenko, M. N., & Semenova, A. N. (2019). HPLC analysis of an ascorbic acid in biological fluids and some foodstuff. Sorbtsionnye I Khromatograficheskie Protsessy, 13(3). Retrieved from https://journals.vsu.ru/sorpchrom/article/view/1627
Issue
Vol 13 No 3 (2013): Sorbtsionnye i Khromatograficheskie Protsessy
Section
Статьи