Development of salt-tolerance interface for an high performance liquid chromatography/inductively coupled plasma mass spectrometry system and its application to accurate quantification of DNA samples.
Accurate quantification of DNA is highly important in various fields. Determination of phosphorus by ICP-MS is one of the most effective methods for accurate quantification of DNA due to the fixed stoichiometry of phosphate to this molecule. In this paper, a smart and reliable method for accurate quantification of DNA fragments and oligodeoxythymidilic acids by hyphenated HPLC/ICP-MS equipped with a highly efficient interface device is presented. The interface was constructed of a home-made capillary-attached micronebulizer and temperature-controllable cyclonic spray chamber (IsoMist). As a separation column for DNA samples, home-made methacrylate-based weak anion-exchange monolith was employed. Some parameters, which include composition of mobile phase, gradient program, inner and outer diameters of capillary, temperature of spray chamber etc., were optimized to find the best performance for separation and accurate quantification of DNA samples. The proposed system could achieve many advantages, such as total consumption for small amount sample analysis, salt-tolerance for hyphenated analysis, high accuracy and precision for quantitative analysis. Using this proposed system, the samples of 20 bp DNA ladder (20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 300, 400, 500 base pairs) and oligodeoxythymidilic acids (dT(12-18)) were rapidly separated and accurately quantified.
Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. takasaki.yuka@g.mbox.nagoya-u.ac.jp
Yuka
Y
Takasaki
Shinnosuke
S
Sakagawa
Kazumi
K
Inagaki
Shin-Ichiro
S
Fujii
Akhmad
A
Sabarudin
Tomonari
T
Umemura
Hiroki
H
Haraguchi
D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D013058:Mass Spectrometry; D009838:Oligodeoxyribonucleotides / Q000032:analysis / Q000302:isolation & purification; D010758:Phosphorus / Q000032:analysis; D055049:Salt Tolerance
D009838:Oligodeoxyribonucleotides; C027903:oligo (dT); D010758:Phosphorus; D004247:DNA