A novel approach for analysis of oligonucleotide–cisplatin interactions by continuous elution gel electrophoresis coupled to isotope dilution inductively coupled plasma mass spectrometry and matrix‐assisted laser desorption/ionization mass spectrometry

Abstract

In this work we present a novel approach for in vitro studies of cisplatin interactions with 8-mer oligonucleotides. The approach is based on the recently developed coupling of continuous elution gel electrophoresis (GE) to an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) with the aim of monitoring the interaction process between this cytostatic drug and the nucleotides. In contrast to existing methods, the electrophoretic separation conditions used here allow both the determination of the reaction kinetics in more detail as well as the observation of dominant intermediates. Two different nucleotides sequences have been investigated for comparison purposes, one containing two adjacent guanines (5'-TCCGGTCC-3') and one with a combination of thymine and guanine (5'-TCCTGTCC-3'), respectively. In order to gain further structural information, MALDI-TOF MS measurements have been performed after fraction collection. This allows for identification of the intermediates and the final products and confirms the stepwise coordination of cisplatin via monoadduct to bisadduct formation. Furthermore, the ICP-MS results were quantitatively evaluated in order to calculate the kinetics of the entire process.

Separation and quantification of RNA molecules using size‐exclusion chromatography hyphenated with inductively coupled plasma‐mass spectrometry

“…Although the determination of phosphorus in a DNA molecule using ICP‐MSor ICP‐optical emission spectrometry (OES) has been recognized as an accurate DNA quantification, the determination results only show the total amount of DNA in the sample . Therefore, hyphenated techniques such as LC or electrophoresis with ICP‐MS are considered useful analytical tools . We have reported the coupling techniques of LC with ICP‐MS, where we employed RP chromatography only for the analysis of mono‐nucleotides was performed .…”

Inductively Coupled Plasma Mass Spectrometry

“…The interaction of cisplatin and guanosine monophosphate was also studied by Hann et al 395 They found mono-and biadducts, which were separated from one another by HPIC and analysed for of 31 396 The artificial oligonucleotides contained either adjacent guanosines (GG) or not (TG). Mono-and biadducts were detected and the respective binding constants could be determined.…”

Quantification of Noble Metals in Biological and Environmental Samples

“…CE offers the possibility of investigating the interaction under conditions corresponding to physiological conditions (aqueous media, pH, and temperature) and is valuable for such studies [146,149,[167][168][169][170][171][172]. Gel electrophoresis coupled with ICP MS and MALDI TOF MS has proved useful for examination of the interaction of cisplatin with oligonucleotides [173].…”