Česká společnost pro hmotnostní spektrometrii

(CEEPC/IPM/CMSC 2022 - FrP-02)
Development of on-line pretreatment methods for growth factor analysis using CZE-MS

Martina Opetová ^1 *, Radovan Tomašovský ¹, Peter Mikuš ^1,2, Katarína Maráková ^1,2

1. Faculty of Pharmacy, Comenius University in Bratislava
2. Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University in Bratislava

Abstrakt

Growth factors are a heterogeneous group of proteins that are secreted by various tissues in the body. Their presence in biological samples may indicate the development of various diseases, so there is a growing need to develop new high-performance analytical methods and procedures for their identification and quantification. The developed methods could subsequently be used either in medicine and diagnostics, or in therapeutic monitoring and anti-doping. The aim of this work is the development of on-line pretreatment methods aimed at concentrating the sample before the analysis itself using an on-line combination of capillary zone electrophoresis (CZE) and mass spectrometry (MS) [1,2]. Optimization and comparison of two pretreatment methods, transient isotachophoresis (tITP) and dynamic pH junction (DPJ), was performed on a mixture of standards of three selected intact growth factors, namely IGF-1 (insulin-like growth factor-1), EGF (epidermal growth factor), and TGF-α (transforming growth factor- α). The lowest detection limits of 66 ng/mL for IGF-1 and EGF and 40 ng/mL for TGF- α were reached using the transient isotachophoresis (tITP) method and using 200 mmol/L ammonium formate (pH 4.0) as the leading electrolyte. The benefit of such a method is a prerequisite for its use in the analysis of growth factors as potential biomarkers of diseases in biological samples, where these substances are found at very low concentration levels.

* Korespondující autor: opetova2@uniba.sk

Literatura

1. Wu, H. et al: Reviews in Analytical Chemistry 39(1), 45-55 (2020).
2. Nickerson, J. L. et al: Mass Spectrometry Reviews (2021).

Poděkování:

This work was supported by the grants VEGA 1/0483/20, VEGA 1/0514/22, UK/93/2022, and UK/110/2022 and carried out in the Toxicological and Antidoping Center at the Faculty of Pharmacy, Comenius University in Bratislava.