Česká společnost pro hmotnostní spektrometrii

(CEEPC/IPM/CMSC 2022 - FrP-08)
Development and optimization of CZE-MS method for the analysis of selected growth factors

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

1. Farmaceutická fakulta Univerzity Komenského v Bratislave

Abstrakt

Growth factors have many physiologically beneficial functions in the body. However, in various diseases, an imbalance between stimulatory and inhibitory signals can occur, which can result in abnormal expression of growth factors [1]. These qualitative and/or quantitative changes can occur much earlier than the disease manifests itself, and the size of the abnormality may indicate disease progression or treatment success. Analysing these changes is still a challenge today and requires the development of new high-efficiency separation and detection methods.

In this work, we optimized the capillary zone electrophoresis (CZE) hyphenated with mass spectrometry (MS) for top-down analysis of insulin-like growth factor-1, transforming growth factor-α and epidermal growth factor.

First, we identified the charge state distribution of each protein and optimized the fragmentor voltage and dwell time for selected ions (m/z). Suitable background electrolyte (BGE) was composed of 500 mM HFo with 5% ACN, and the optimum separation voltage was set at 20 kV. Next, we compared bare fused-silica capillary with permanently coated polyvinylalcohol capillary and linear polyacrylamide capillary. The optimal sheath liquid consisted of 50% MeOH with 0.1% HFo. The drying gas was set at a temperature of 300 °C and a flow rate of 6 L/min. Nebulizing gas pressure was set at 4 psi and capillary voltage at 4 kV.

With optimized parameters, we reached preliminary LOD of 250 µg/L for all three analytes. Further optimization is needed before the method will be suitable for the analysis of growth factors also in biological samples.

* Korespondující autor: tomasovsky2@uniba.sk

Literatura

1. P. H. Gann, R. T. Chatterton, C. Lee, Epidemiologic Reviews. 23, 67–71 (2001).

Poděkování:

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