NIST logo

Publication Citation: Potential Primary Measurement Tool for the Quantification of DNA

NIST Authors in Bold

Author(s): Ryan G. Brennan; Savelas A. Rabb; Marcia J. Holden; Michael R. Winchester;
Title: Potential Primary Measurement Tool for the Quantification of DNA
Published: May 01, 2009
Abstract: An automated sample introduction system, utilizing a demountable direct injection high efficiency nebulizer (d DIHEN), is successfully incorporated for the first time with an inductively coupled plasma optical emission spectrometer (ICP-OES) for the measurement of the phosphorus content in acid-digested nucleotides and deoxyribonucleic acid (DNA). With this experimental setup, the solution uptake rate is reduced from 170 L min-1 to 30 L min-1, and the required sample size is reduced from 10 mL to 2.4 mL in comparison to previous analyses in our lab using a glass, concentric nebulizer with cyclonic spray chamber arrangement. The use of direct injection improves P (I) 213.617 nm sensitivity by a factor of 4 on average. A high performance (HP) methodology in combination with the current sample introduction system and ICP-OES provides simultaneous, time-correlated internal standardization and drift correction resulting in relative expanded uncertainties (% U) for the P mass fractions in the range of 0.1 to 0.4 (95 % confidence level) for most of the thymidine 5 -monophosphate (TMP), calf thymus DNA (CTDNA), and plasmid DNA (PLDNA) analyses. The d-DIHEN with HP-ICP-OES methodology allows for the quantification of DNA mass at P mass fractions as low as 0.5 g g-1 with small uncertainty ( 0.4 %). This successful approach will aide in the development and certification of nucleic acid certified reference materials (CRMs), particularly for these samples that are typically limited in volume
Citation: Analytical Chemistry
Volume: 91
Pages: 9 pp.
Keywords: ICP-OES, DNA, d-DIHEN, Phosphorous
Research Areas: Chemistry
PDF version: PDF Document Click here to retrieve PDF version of paper (1MB)