A comparison of enzymatic digestion for the quantitation of an oligonucleotide by liquid chromatography–isotope dilution mass spectrometry

Analysis of mono-phosphate nucleotides as a potential method for quantification of DNA using high performance liquid chromatography–inductively coupled plasma-mass spectrometry

“…Elemental analysis is preferred over other techniques as it offers lower detection limits, a larger dynamic range, a response that is to a first approximation independent of molecular form, and potentially more accurate quantification at low concentrations by isotope dilution analysis. Both ICP-MS and ICP-OES have been utilised for the quantification of nucleotides, DNA, oligonucleotides and DNA adducts [3][4][5][6][7][8][9][10][11][12][13][14][15]. The quantification of oligonucleotides has been achieved by two different methods by Yang et al [4] and Donald et al [7], with the latter group digesting the oligonucleotides to mono-nucleosides and quantifying by isotope dilution electrospray mass spectrometry (ES-ID-MS), and comparing that value to the total 31 P signal obtained from ICP-OES and the gravimetric value.…”

“…Both ICP-MS and ICP-OES have been utilised for the quantification of nucleotides, DNA, oligonucleotides and DNA adducts [3][4][5][6][7][8][9][10][11][12][13][14][15]. The quantification of oligonucleotides has been achieved by two different methods by Yang et al [4] and Donald et al [7], with the latter group digesting the oligonucleotides to mono-nucleosides and quantifying by isotope dilution electrospray mass spectrometry (ES-ID-MS), and comparing that value to the total 31 P signal obtained from ICP-OES and the gravimetric value. Styrene oxidate and mephalen DNA adducts have been investigated by elemental mass spectrometry, by Edler et al [12,13] and styrene -7,8-oxide adducts by Siethoff et al [11].…”

Development of Metrology for Modern Biology

“…Nucleotide could be the quantification target (O'Connor et al, 2002) whereas nucleoside can be also a measurement target (Donald et al, 2005). Phosphodiesterase produces single nucleotide from a DNA sequence with its activity for hydrolysis of phosphodiester bonds at the 5'-position.…”

“…O'Connor et al first reported the use of HPLC-IDMS for accurate quantitation of nucleotides resulted from enzymatic hydrolysis of a DNA sequence (O'Connor et al, 2002). Later, they reported that analysis of the nucleosides instead of nucleotides resulted in better estimation of the quantity of the target DNA (Donald et al, 2005). For both analytical targets, isotope dilutionmass spectrometry (ID-MS) was applied to secure metrological quality of analysis.…”

Quantification of an Oligonucleotide Containing a Sequence Failure Product: Comparison of Isotope Dilution Mass Spectrometry with other Quantification Methods

“…the ID MS results were compared with those obtained by other analytical methods (figure 1). the ID MS-based method performed quantification of constituent deoxynucleotide monophosphates (dNMPs) and deoxynucleosides (dNs) 6 after enzymatic digestion of the oligonucleotide. We compared these results with those of IcP-oES for quantification of the constituent phosphorus of the oligonucleotide, ion chromatography for quantification of the phosphate after enzymatic digestion of the oligonucleotide, uV absorption and gravimetric methods.…”