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A novel targeted analysis of peripheral steroids by ultra–performance supercritical fluid chromatography hyphenated to mass spectrometry
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Kemiska sektionen, Institutionen för kemi - BMC, Analytisk kemi. (Bergquist group)ORCID-id: 0000-0002-7167-5473
(engelsk)Inngår i: Artikkel i tidsskrift (Annet vitenskapelig) Submitted
HSV kategori
Forskningsprogram
Kemi med inriktning mot analytisk kemi
Identifikatorer
URN: urn:nbn:se:uu:diva-356766OAI: oai:DiVA.org:uu-356766DiVA, id: diva2:1236899
Tilgjengelig fra: 2018-08-06 Laget: 2018-08-06 Sist oppdatert: 2018-08-06
Inngår i avhandling
1. Mass spectrometry based analysis of endogenous sterols and hormones
Åpne denne publikasjonen i ny fane eller vindu >>Mass spectrometry based analysis of endogenous sterols and hormones
2018 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Bioanalytical applications using supercritical fluid chromatography (SFC) as analytical technique are of increasing interest. In essence, bioanalysis involves measurement of bioactive or endogenous compounds in biological matrices. SFC has emerged as an excellent choice for bioanalytical analysis, attributable to its speed, selectivity and efficiency compared with high performance liquid chromatography. Moreover, coupling of SFC with mass spectrometry (MS) provides the additional benefits of specificity and sensitivity.

The aim of this thesis was to exploit these features by developing methods for the analysis of endogenous steroids, cholesterol oxidation products (COPs) and thyroid hormones (THs) by using ultra-performance supercritical fluid chromatography–tandem mass spectrometry (UPSFC–MS/MS) as analytical technique.

Endogenous steroids control many physiological processes, including reproduction, maturation, gene expression and neurological functions in humans and animals. In the first study, three steroids were measured in domesticated White Leghorn (WL) chickens and ancestral Red Junglefowl (RJF) birds. Restraining stress caused a significantly larger increase in corticosterone levels in RJF than in WL, indicating a blunted hypothalamic–pituitary–adrenal (HPA) axis activity in domesticated chickens. The second study was a continuation of the first study and corticosterone levels from the F12 generation of an intercross between WL and RJF birds were measured before and after physical restraint stress. The expression levels of the glucocorticoid receptor (GR) in the hypothalamus and several genes in the adrenal glands were correlated with the post-stress levels of corticosterone in plasma. In the third study, the measurement of steroids was extended to assess more endogenous steroids from the four major classes, i.e. estrogens, androgens, progestogens and corticosterone.

Endogenous COPs are of interest in pathophysiology. COPs are more readily disposed by cells than cholesterol. Therefore, cholesterol is oxidised to the more polar COPs and are generally more bioactive than cholesterol. Moreover, if their production in cells and tissues and/or their introduction with dietary animal fat are excessive, COPs could indeed contribute to the pathogenesis of various disease processes. Fourteen COPs were included in the fourth study and a novel method for their separation was developed.

The last study in this thesis, involved the analysis of five THs. These hormones are vital for growth, developmental and metabolic processes of vertebrate life and play an important role in energy homeostasis. Measurements of circulating thyroid hormone levels are used in thyroid disorder diagnoses or treatment status monitoring. Two rapid methods for the separation of five THs were developed.

In summary, the work in this thesis demonstrates the applicability of UPSFC–MS/MS as an analytical technique in bioanalysis of endogenous compounds.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis, 2018. s. 62
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1698
Emneord
mass spectrometry, supercritical fluid chromatography, UPSFC–MS/MS, bioanalysis, steroids, oxysterols, thyroid hormones
HSV kategori
Forskningsprogram
Kemi med inriktning mot analytisk kemi
Identifikatorer
urn:nbn:se:uu:diva-356767 (URN)978-91-513-0396-3 (ISBN)
Disputas
2018-09-19, B7:101a, BMC, Husargatan 3, Uppsala, 09:15 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2018-08-30 Laget: 2018-08-06 Sist oppdatert: 2018-09-10

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