Probing the Chemistry of Methylmercury in Mammalian Blood Plasma

Bridle, Tristen G.

Probing the Chemistry of Methylmercury in Mammalian Blood Plasma

dc.contributor.advisor	Gailer, Jürgen
dc.contributor.author	Bridle, Tristen G.
dc.contributor.committeemember	Jalilehvand, Farideh
dc.contributor.committeemember	Thurbide, Kevin
dc.date	Winter Conferral
dc.date.accessioned	2022-11-15T17:43:29Z
dc.date.embargolift	2022-07-04
dc.date.issued	2022-01-04
dc.description.abstract	A size-exclusion chromatography-inductively coupled plasma-atomic emission spectroscopy (SEC-ICP-AES) technique was developed for the analysis of mammalian blood plasma for endogenous metalloproteins and exogenous metal species. Parameters related to sample preparation, separation, and detection were optimized. The blood plasma was filtered through a 0.45 ?m pore size syringe filter to preserve the separation performance of the column. The 1:1 dilution of filtered human and rabbit blood plasma in phosphate-buffered saline (PBS) prior to analysis was needed to prevent peak tailing and shoulder formation. An SRT-10C SEC column from Sepax Technologies Inc., when compared to a previously used Superdex 200 Increase SEC column, gave the same number of peaks with similar resolution, but in approximately half the time and with higher peak intensities. The mass recovery of proteins on the SRT-10C SEC column was 96 ± 2%. To explore the role that small molecular weight plasma thiols play in the delivery of methylmercury (CH3Hg+) to transporters located at the placental and blood-brain barriers, the described SEC-ICP-AES technique was applied to the analysis of CH3Hg+-spiked rabbit plasma using PBS mobile phases in the absence and presence of the small-molecular weight (SMW) sulfur compounds L-cysteine (Cys), L-homocysteine (hCys), L-glutathione (GSH), and D-methionine (Met). While Met did not affect the binding of CH3Hg+ to the main plasma protein, rabbit serum albumin (RSA), Cys, hCys, and GSH did. The presence of 50 ?M Cys, hCys, or GSH in the mobile phase resulted in the mobilization of CH3Hg+ from RSA in rabbit plasma and from pure RSA in solution. The SMW-Hg species that formed when hCys was present in the mobile phase was qualitatively identified by electrospray ionization mass spectrometry as CH3Hg-hCys. Using the developed metallomics technique I have found evidence of the formation of SMW-CH3Hg+ species under near-physiological conditions that may be involved in the translocation of CH3Hg+ from blood plasma to the brain.
dc.identifier.citation	Bridle, T. G. (2022). Probing the Chemistry of Methylmercury in Mammalian Blood Plasma (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.
dc.identifier.uri	http://hdl.handle.net/1880/115495
dc.identifier.uri	https://dx.doi.org/10.11575/PRISM/40462
dc.language.iso	en	en
dc.language.iso	English
dc.publisher.faculty	Graduate Studies	en
dc.publisher.faculty	Science
dc.publisher.institution	University of Calgary	en
dc.rights	University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.	en
dc.subject	size-exclusion chromatography
dc.subject	inductively coupled plasma
dc.subject	atomic emission spectroscopy
dc.subject	methylmercury
dc.subject	blood plasma
dc.subject	thiol
dc.subject	cysteine
dc.subject	homocysteine
dc.subject	glutathione
dc.subject	metalloprotein
dc.subject.classification	Chemistry--Analytical
dc.subject.classification	Chemistry--Biochemistry
dc.subject.classification	Chemistry--Inorganic
dc.title	Probing the Chemistry of Methylmercury in Mammalian Blood Plasma
dc.type	master thesis
thesis.degree.discipline	Chemistry
thesis.degree.grantor	University of Calgary	en
thesis.degree.grantor	University of Calgary
thesis.degree.name	Master of Science (MSc)