Potential Primary Measurement Tool for the Quantification of DNA

Published

May 1, 2009

Author(s)

Ryan G. Brennan, Savelas A. Rabb, Marcia J. Holden, Michael R. Winchester

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

Pub Type

Journals

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Keywords

ICP-OES, DNA, d-DIHEN, Phosphorous

Citation

Brennan, R. , Rabb, S. , Holden, M. and Winchester, M. (2009), Potential Primary Measurement Tool for the Quantification of DNA, Analytical Chemistry, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=832413 (Accessed December 26, 2024)