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The Electrophoretic Determination oF Biological Active Substances

#3850


The Strategy of the Сhoice of Analytical Method for Determination of Bioactive Substances (Steroids, Neurotransmitters, Proteins and Natural Antioxidants) in Biological Objects (Urine, Blood, Tears at al.)

Tech Area / Field

  • CHE-ANL/Analytical Chemistry/Chemistry
  • BIO-CHM/Biochemistry/Biotechnology
  • MED-DIS/Disease Surveillance/Medicine

Status
3 Approved without Funding

Registration date
21.02.2008

Leading Institute
St Petersburg State University, Russia, St Petersburg

Supporting institutes

  • Saint Petersburg State Polytechnical University, Russia, St Petersburg

Collaborators

  • Universita degli Studi del Sannio, Italy, Benevento\nUniversité du Québec en Outaouais, Canada, QC, Gatineau\nPlasmionique Inc., Canada, Varennes\nWarsaw University of Technology / Institute of Microelectronics and Optoelectronics, Poland, Warsaw

Project summary

The aim of the project is developing an algorithm and methodology of rapid and highly efficient chromatographic and electrophoretic determination of bio-active substances (biogenic amines, polyphenols, proteins, steroids etc.) in biological objects for endocrine, cardiovascular, tumorous, neuro and mental diseases diagnosing an early stage. This algorithm is based on the molecular recognition principle.

Methods of investigation:

  • Reversed-phase high performance liquid chromatography with ultraviolet, fluorescence and electrochemical detection (HPLC-UV, HPLC-ED);
  • High performance thin layer chromatography with video and UV densitometry (HPTLC-UV);
  • Capillary electrophoresis (capillary zone electrophoresis (CZE); micelle electrokinetic chromatography (MEKC) with ultraviolet (UV) and mass spectrometric (MS) detections;
  • Mass spectrometry (MALDI-TOF MS).

Objects of research: urine, plasma and blood serum, saliva, cerebrospinal liquid, tears.

The measurement and monitoring of bio-active substances maintenance (catecholamines and their metabolites, protein (albumin, globulin), steroids hormone, etc.) in human physiological liquids is an actual point and is of great practical value for clinical diagnosis of various diseases (Parkinson's disease, pheochromocytoma, neuroblastoma, schizophrenia, epilepsy, cardiovascular disease). In case of brain functions disorder tumor, or during neural and mental diseases their concentration varies in different biological objects.

Specific topics of scientific investigations within the project:

  • Study of the effect of adding complexing agents (crown-ether, cyclams, ion-pair additive, micelles, cation metals etc.) to the mobile phase mixture or background electrolyte on the selectivity and efficiency of electrophoretic and chromatographic separation of bio-active compound model solutions containing carboxy-, hydroxy- and amino groups. Quantitative estimation and investigation of the role of complex forming processes in selective determination of bio-active substances;
  • Research of new methods of on-line enhanced sample injection alongside improved with ones. Study of normal stacking, stacking with reversed migration micelles, stacking with high conductivity matrix (with use of additive NaCl), reversed migration micelles and sweeping etc. Study of off-line concentration using organized mediums (micelles, macrocycles, microemulsions, urea, etc.) with the aim of reducing the detection limits of important diagnostic analytes and enhancing the UV-detection sensitivity;
  • Development of sample preparation procedure of biological liquids (urine, plasma, blood serum, tears, saliva) for electrophoretic and chromatographic determination of neurotransmitters and their metabolites;

The main part of our scientific team are qualified specialists, young, but experienced in determining biological active substances (amines, amino- and hidroxy- acids, steroids, proteins and so on) – diagnosing markers of various diseases – by chromatographic and electrophoretic methods with using comlexing agents (crown-ethers, criptands, cyclam, cyclodextrin and his derivates, ion-pair additive) in composition of eluent or background electrolyte; in quantitative estimation of interaction in system analyte – complexing agent; in development sample preparation of biological liquids and plants with using different way of on-line and off-line concentrations (concentration factors is 700 – 1000).

The results of our research have been published in refereed scientific journals and presented at many chemistry and medicine international conferences in Russia, Italy, Spain, Germany, Poland, China, and Norway.

International collaborators from Canada, Poland and Italy actively participate in the project. All experts involved are acknowledged ones. Their scientific interests are complementary to those of the Russian members of the team. Those interests involve capillary electrophoresis, fiber optic sensors and sensing technologies, analysis and recognition of liquids using optical capillaries, biotechnology, plasma chemistry, physics and instrumentation, computer modeling using neural networks and other techniques.


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