Quantitation of Normal and Formaldehyde-Modified Deoxynucleosides by High-Performance Liquid Chromatography/UV Detection

Details

• Personal Author:
  Que Hee S ; Zhong W
• Description:
  A sensitive and selective method was developed for the first time to quantify simultaneously the normal and formaldehyde (FA)-modified bases in human placental DNA treated with 100 ppm FA for 20 h at 37 degrees C. Digestion of DNA to deoxynucleosides with DNase I, phosphodiesterase and alkaline phosphatase occurred in that order with centrifugation steps. The normal and FA-modified deoxynucleosides were then resolved from one another and reagent blank interferences to produce selective separation through high performance liquid chromatography-ultraviolet detection at 254 nm. A C18 reversed-phase column facilitated the resolution using 5 mm ammonium acetate and a gradient of 0-6% methanol at flow rates of 0.3-1.4 mL/min before column cleaning. The lower quantifiable limits for deoxyadenosine, deoxyguanosine, deoxycytidine, thymidine, N6-hydroxymethyldeoxyadenosine (N6-dA), N2-hydroxymethyldeoxyguanosine (N2-dG) and N4-hydroxymethyldeoxycytidine (N4-dC) were 11, 7.6, 12, 15, 10, 10 and 22 pmol, respectively. The abundance order of the modified deoxynucleosides was N6-dA > N2-dG > N4-dC. dT did not form hydroxymethyl derivatives. The respective concentrations were about 6.0, 10.0 and 23 pmol of modified deoxynucleosides in 80 ug of human placental DNA after treatment with 100 µg/mL of formalin for 20 h at 37 degrees C. The stabilities of N6-dA and N2-dG were much better at -20 degrees C than at 25 degrees C, where the respective halftimes were about 50.1 and 21.0 h. [Description provided by NIOSH]
• Subjects:
  Formaldehyde Laboratories Occupational Health Safety Temperature
• Keywords:
  Author Keywords: DNA Adducts Biomarkers Deoxyribonucleic Acids DNA HPLC/UV Hydroxymethyldeoxynucleosides Laboratory Techniques Liquid Chromatography Temperature Effects
• ISSN:
  0269-3879
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  California ; OSHA Region 9
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  462-469
• Volume:
  18
• Issue:
  7
• NIOSHTIC Number:
  nn:20057949
• Citation:
  Biomed Chromatogr 2004 Sep; 18(7):462-469
• Contact Point Address:
  W. Zhong, Department of Environmental Health, University of California, Los Angeles, 650 Charles Young Drive South, Los Angeles, California 90095-1772, USA
• Email:
  wzhong@ucla.edu
• CAS Registry Number:
  Formaldehyde (CAS RN 50-00-0)
• Federal Fiscal Year:
  2004
• Performing Organization:
  University of California, School of Public Health, Los Angeles, CA
• Peer Reviewed:
  True
• Start Date:
  19990701
• Source Full Name:
  Biomedical Chromatography
• End Date:
  20040630
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:f8e907cb74d33040d2e8b5d6c6098c51821f293137b5418122de53471df3aaf41552876e352db2be956e05e8132a1ea2b5e44fdf040e39130a8b3a127694fe69
• Download URL:
  https://stacks.cdc.gov/view/cdc/215151/cdc_215151_DS1.pdf
• File Type:
  [PDF - 125.48 KB ]