The detection and quantification of biomolecules in biological sample matrices

Abstract

In the bodies of living humans and animals, endogenous biomolecules, and in particular the chemical messengers, play an essential role in the maintenance and functionality of physiological processes (Daintith, 2008; Gault & McClenaghan, 2013). When these chemical messengers are out of balance, however, it can cause a variety of disorders throughout the whole body. The analytical measurement of these biomolecules can play an essential role in our understanding of both the normal physiological and the abnormal pathophysiological processes. This understanding can help with better treatment of these disorders and development of newer and better drugs. In the research environment, costs related to analysis of biomolecules of either chemical messengers or biomarkers for certain disorders is a problem. The current commercial available radioimmunoassay (RIA) (Wassell et al., 1999), enzyme immunoassay (EIA) (Kemppainen et al., 2018) and enzyme-linked immunosorbent assay (ELISA) (Kim et al., 2008) kits are expensive per sample and because of cross contamination, some are unreliable. Chromatography is an analytical technique for the separation of mixtures of compounds and molecules in solutions by a variety of different chemical processes (Licker, 2003), and a variety of sample preparation techniques. The aim of this study was to develop and validate new methods for the analysis of endogenous biomolecules that would be sensitive, specific, reliable and affordable. We developed and validated three new methods, which were published in an international accredited journal. The first method was for the analysis of the monoamines and their metabolites in rat brain tissue. The second was for norepinephrine and its metabolites for the measurement of enzyme COMT activity in rat liver homogenate in the presence of the known COMT inhibitor, entacapone. Lastly, for the analysis of cortisol, corticosterone and melatonin in plasma samples from laboratory animals, the Sprague-Dawley rat and the white rhinoceros, a wild animal. These methods not only adhered to the aims and objectives of the study project but also to the research problem of the need to develop and validate new analytical methods that would be sensitive, specific, reliable and more affordable than commercially available analytical kits. We conclude that these newly developed and validated analytical methods can be applied in practice with great success and with cost saving advantages. We anticipate that these methods will be a great addition especially to our research environment, where these kind of methods are constantly used in a variety of animal behavioural studies in stress or stress-related disorders. These methods can also be used for better drug development and discovery.