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Oligonucleotide analysis by MALDI-ion-mobility-TOFMS (CAT#: STEM-ST-0103-LJX)

Introduction

Matrix-assisted laser-desorption ionization followed by ion-mobility separation and time-of-flight mass analysis (MALDI-IM-TOFMS) has been used to characterize native and chemically modified DNA oligonucleotides up to eight bases in length. Mobility resolution between 20 and 30 can be used to separate oligonucleotides of different length, but not to differentiate between isomers or even different compositions of the same length. MALDI-IM-TOFMS does, however, have additional utility in the analysis of mixtures of DNA oligonucleotides and peptides, because these classes of molecules can be distinguished on the basis of differences in their mobility. Oligonucleotide sequencing is also possible by MALDI-IM-TOFMS. Ion signals corresponding to nucleobase losses, w-type, and y-type fragments were identified by use of differences in ion mobility. MALDI-IM-TOFMS was also used to resolve DNA-platinum adducts from the corresponding unmodified oligonucleotides.




Principle

Ion mobility spectrometry–mass spectrometry (IMS-MS) is an analytical chemistry method that separates gas phase ions based on their interaction with a collision gas and their masses. In the first step, the ions are separated according to their mobility through a buffer gas on a millisecond timescale using an ion mobility spectrometer. The separated ions are then introduced into a mass analyzer in a second step where their mass-to-charge ratios can be determined on a microsecond timescale.

Applications

For studying the gas phase ion structure
For detecting the chemical warfare agents and explosives
For the analysis of proteins, peptides, drug-like molecules and nano particles
For monitoring isomeric reaction intermediates and probe their kinetics
For proteomics and pharmaceutical analysis

Procedure

1. Add sample
2. The ions in the sample are separated in the ion mobility spectrometer
3. The separated ions are introduced into the mass analyzer for detection
4. Store the detection results

Materials

• Sample Type:
Oligonucleotide

Notes

1. Ion mobility spectrometry is also a very fast technique, making it suitable for high-throughput applications. The entire analysis can be completed in just a few minutes.
2. The method is extremely sensitive and able to detect trace amounts of contaminants that other spectrometry methods would miss.
3. The effective separation of analytes achieved with this method makes it widely applicable in the analysis of complex samples such as in proteomics and metabolomics.
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