title: Simultaneous Measurement of 3-Chlorotyrosine and 3,5-Dichlorotyrosine in Whole Blood, Serum and Plasma by Isotope Dilution HPLC-MS-MS.
abstract: Chlorine is a public health concern and potential threat due to its high reactivity, ease and scale of production, widespread industrial use, bulk transportation, massive stockpiles and history as a chemical weapon. This work describes a new, sensitive and rapid stable isotope dilution method for the retrospective detection and quantitation of two chlorine adducts. The biomarkers 3-chlorotyrosine (Cl-Tyr) and 3,5-dichlorotyrosine (Cl2-Tyr) were isolated from the pronase digest of chlorine exposed whole blood, serum or plasma by solid-phase extraction (SPE), separated by reversed-phase HPLC and detected by tandem mass spectrometry (MS-MS). The calibration range is 2.50-1,000 ng/mL (R2 ≥ 0.998) with a lowest reportable limit (LRL) of 2.50 ng/mL for both analytes, an accuracy of ≥93% and an LOD of 0.443 ng/mL for Cl-Tyr and 0.396 ng/mL for Cl2-Tyr. Inter- and intra-day precision of quality control samples had coefficients of variation of ≤10% and ≤7.0%, respectively. Blood and serum samples from 200 healthy individuals and 175 individuals with chronic inflammatory disease were analyzed using this method to assess background levels of chlorinated tyrosine adducts. Results from patients with no known inflammatory disease history (healthy) showed baseline levels of <LRL-4.26 ng/mL Cl-Tyr and <LRL Cl2-Tyr. Patients with inflammatory disease had baseline levels of <LRL-15.4 ng/mL Cl-Tyr and <LRL-5.22 ng/mL Cl2-Tyr. Blood exposed to 2.02 ppm chlorine gas for 15 min produced 941 ng/mL Cl-Tyr and 223 ng/mL Cl2-Tyr. This high-throughput method has been developed and analytically validated for the diagnosis of human exposure to chlorine.
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA jgz8@cdc.gov.
authors.forename: Brian S
authors.initials: BS
authors.lastname: Crow
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Quiñones-González
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Brooke G
authors.initials: BG
authors.lastname: Pantazides
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Jonas W
authors.initials: JW
authors.lastname: Perez
authors.affiliation: Battelle Memorial Institute, Atlanta, GA 30329, USA.
authors.forename: W Rucks
authors.initials: WR
authors.lastname: Winkeljohn
authors.affiliation: Oak Ridge Institute for Science and Education Fellow at the Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Joshua W
authors.initials: JW
authors.lastname: Garton
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Jerry D
authors.initials: JD
authors.lastname: Thomas
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Blake
authors.affiliation: Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
authors.forename: Rudolph C
authors.initials: RC
authors.lastname: Johnson
mesh_terms: D015415:Biomarkers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D007249:Inflammation / Q000652:urine; D057230:Limit of Detection; D010949:Plasma / Q000737:chemistry; D011786:Quality Control; D011865:Radioisotope Dilution Technique; D015203:Reproducibility of Results; D012189:Retrospective Studies; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000097:blood
chemical_list: D015415:Biomarkers; D007202:Indicators and Reagents; D014443:Tyrosine; C408691:3,5-dichlorotyrosine; C087259:3-chlorotyrosine