title: Mass-spectrometric profiling of porphyrins in complex biological samples with fundamental, toxicological, and pharmacological applications.
abstract: Rapid, high-throughput, and quantitative evaluations of biological metabolites in complex milieu are increasingly required for biochemical, toxicological, pharmacological, and environmental analyses. They are also essential for the development, testing, and improvement of new commercial chemical products. We demonstrate the application of ultra-high performance liquid chromatography-mass spectrometry (uHPLC-MS), employing an electrospray ionization source and a high accuracy quadrupole time-of-flight mass analyzer, for the identification and quantification of a series of porphyrin derivatives in liver: a matrix of particular relevance in toxicological or pharmacological testing. Exact mass is used to identify and quantify the metabolites. Chromatography enhances sensitivity and alleviates potential saturation issues by fanning out the contents of a complex sample before their injection into the spectrometer, but is not strictly necessary for the analysis. Extraction and sample treatment procedures are evaluated and matrix effects discussed. Using this method, the known mechanism of action of a well-characterized porphyrinogenic agent was verified in liver extracts from treated rats. The method was also validated for use with bacterial cells. This exact-mass method uses workhorse instruments available in many laboratories, providing a highly flexible alternative to existing HPLC- and MS/MS-based approaches for the simultaneous analysis of multiple compounds in biological media.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Sullivan
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA; Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59713, USA.
authors.forename: Bennett R
authors.initials: BR
authors.lastname: Streit
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Ethan L
authors.initials: EL
authors.lastname: Ferguson
authors.affiliation: Health and Environmental Sciences, Dow Corning Corporation, Auburn, MI 48611, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Jean
authors.affiliation: Health and Environmental Sciences, Dow Corning Corporation, Auburn, MI 48611, USA.
authors.forename: Debra A
authors.initials: DA
authors.lastname: McNett
authors.affiliation: Health and Environmental Sciences, Dow Corning Corporation, Auburn, MI 48611, USA. Electronic address: louis.t.llames@dowcorning.com.
authors.forename: Louis T
authors.initials: LT
authors.lastname: Llames
authors.affiliation: Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59713, USA. Electronic address: jdubois@chemistry.montana.edu.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: DuBois
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D008099:Liver / Q000378:metabolism*; D011166:Porphyrins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D053719:Tandem Mass Spectrometry
chemical_list: D011166:Porphyrins
keywords: Biomarker; High mass accuracy mass spectrometry; Metabolomic; Porphyrin; Toxicology