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Quantitation of methylated hemoglobin adducts in a signature peptide from rat blood by liquid chromatography/negative electrospray ionization tandem mass spectrometry
Author(s) -
Zhang Fagen,
Bartels Michael J.,
Pottenger Lynn H.,
Schisler Melissa R.,
Grundy Joy J.,
Gollapudi B. Bhaskar
Publication year - 2008
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3530
Subject(s) - chemistry , chromatography , hemoglobin , electrospray ionization , peptide , tandem mass spectrometry , mass spectrometry , adduct , liquid chromatography–mass spectrometry , detection limit , selected reaction monitoring , quantitative analysis (chemistry) , biochemistry , organic chemistry
Hemoglobin adducts are often used as biomarkers for exposure to reactive chemicals in toxicology studies. Therefore, fast, sensitive, accurate, and reproducible methods for quantifying these protein adducts are key to evaluate test material dosimetry. A methodology has been developed for the quantitation of methylated hemoglobin adducts isolated from rats exposed to the model alkylating agent: methyl methane sulfonate (MMS). After 4 days of MMS exposure by oral gavage, hemoglobin was isolated from rat blood and digested with trypsin. The tryptic digestion solution was used for the adducted hemoglobin signature peptide quantitation via liquid chromatography/negative tandem mass spectrometry (LC/ESI‐MS/MS). The limit of quantitation (LOQ) for the methylated hemoglobin beta chain N‐terminal signature peptide (MeVHLTDAEK) was 1.95 ng/mL (5.9 pmol/mg globin). The calibration curves were linear over a concentration range of 1.95 to 625 ng/mL, with a correlation coefficient R 2 >0.998, accuracy of 85.8 to 119.3%, and precision of 0.9 to 19.4%. Copyright © 2008 John Wiley & Sons, Ltd.