HPLC analysis of proline and 4-hydroxyproline in biological fluids

Authors

  • 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
  • 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
  • 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
  • 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 physician ordinator Department of Neurology, Medical Academy, Chita
  • Alena V. Yermolina physician ophthalmologist Regional Hospital № 2, Chita
  • Joseph S Pinelis MD, professor, head of dental surgery Medical Academy, Chita

Keywords:

HPLC, Fluorimetric detection, proline, 4-hydroxyproline, plasma, saliva, plaintive liquid (tear).

Abstract

An HPLC method for measuring proline and 4-hydroxyproline in human plasma, saliva and plaintive liquid
(or tear) was validated. Biological samples were deproteinized using acetonitrile. Iminoacids derivatization was
achieved with 9-fluorenylmethyl chloroformate (Fmoc-Cl) after blocking primary amino acids with
orthophthaldialdehyde/2-mercaptoethanol. The derivatives were separated by reversed phase (C18) HPLC and
fluorimetric detection at ex260-em330 nm. We are used columns Luna C18(2) (75 ´ 4.6 mm, 3 μm), Gemini C18
(150 ´ 4.6 mm, 5 mm), both from Phenomenex and high-speed columns (100 ´ 4.6 mm) Chromolith Performance
RP-18e from Merck. For screening on hyperprolinemia in newborn proposed separation method for high-speed
columns with monolithic sorbent Chromolith Performance RP-18e. 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 screening on congenital hyperprolinemia at newborn

Downloads

Download data is not yet available.

References

Downloads

Published

2019-11-20

Issue

Vol. 13 No. 2 (2013): Sorbtsionnye i Khromatograficheskie Protsessy

Section

Статьи

How to Cite

HPLC analysis of proline and 4-hydroxyproline in biological fluids. (2019). Sorbtsionnye I Khromatograficheskie Protsessy, 13(2). https://journals.vsu.ru/sorpchrom/article/view/1607

Most read articles by the same author(s)