title: Benzoyl chloride derivatization with liquid chromatography-mass spectrometry for targeted metabolomics of neurochemicals in biological samples.
abstract: Widely targeted metabolomic assays are useful because they provide quantitative data on large groups of related compounds. We report a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method that utilizes benzoyl chloride labeling for 70 neurologically relevant compounds, including catecholamines, indoleamines, amino acids, polyamines, trace amines, antioxidants, energy compounds, and their metabolites. The method includes neurotransmitters and metabolites found in both vertebrates and insects. This method was applied to analyze microdialysate from rats, human cerebrospinal fluid, human serum, fly tissue homogenate, and fly hemolymph, demonstrating its broad versatility for multiple physiological contexts and model systems. Limits of detection for most assayed compounds were below 10nM, relative standard deviations were below 10%, and carryover was less than 5% for 70 compounds separated in 20min, with a total analysis time of 33min. This broadly applicable method provides robust monitoring of multiple analytes, utilizes small sample sizes, and can be applied to diverse matrices. The assay will be of value for evaluating normal physiological changes in metabolism in neurochemical systems. The results demonstrate the utility of benzoyl chloride labeling with HPLC-MS/MS for widely targeted metabolomics assays.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, United States.
authors.forename: Jenny-Marie T
authors.initials: JM
authors.lastname: Wong
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, United States.
authors.forename: Paige A
authors.initials: PA
authors.lastname: Malec
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, United States; Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, United States.
authors.forename: Omar S
authors.initials: OS
authors.lastname: Mabrouk
authors.affiliation: Department of Cellular and Molecular Biology Program, University of Michigan, Ann Arbor, MI 48109, United States.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Ro
authors.affiliation: Deparment of Molecular, Cellular, Developmental Biology, University of Michigan, Ann Arbor, MI 48109, United States.
authors.forename: Monica
authors.initials: M
authors.lastname: Dus
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, United States; Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, United States. Electronic address: rtkenn@umich.edu.
authors.forename: Robert T
authors.initials: RT
authors.lastname: Kennedy
mesh_terms: D000596:Amino Acids / Q000032:analysis / Q000134:cerebrospinal fluid; D000818:Animals; D001565:Benzoates / Q000737:chemistry*; D002395:Catecholamines / Q000032:analysis / Q000097:blood / Q000134:cerebrospinal fluid; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004330:Drosophila; D006458:Hemolymph / Q000737:chemistry; D006801:Humans; D055442:Metabolome; D055432:Metabolomics; D018377:Neurotransmitter Agents / Q000032:analysis* / Q000097:blood / Q000134:cerebrospinal fluid; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000596:Amino Acids; D001565:Benzoates; D002395:Catecholamines; D018377:Neurotransmitter Agents; C013409:benzoyl chloride
keywords: Brain; Derivatization; Drosophila; HPLC; Mass spectrometry; Neurochemistry