Separation of phosphorothioated oligonucleotide diastereomers by multiplexed drift tube ion mobility mass spectrometry (CAT#: STEM-ST-0113-LJX)

Introduction

Hydrophilic interaction liquid chromatography (HILIC) coupled to drift tube ion mobility spectrometry (DTIMS) was used to separate diastereomers of five-unit oligonucleotides containing 0, 1, 2 or 3 phosphorothioate (PS) linkages. Multiplexed DTIMS (where ions are pulsed into the drift tube according to a pre-encoded sequence) and post-acquisition processing using an innovative demultiplexing tool were investigated. The electric field inside the drift tube was optimized to achieve the highest resolving power.

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Principle Applications Procedure Materials Notes

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:
Phosphorothioated oligonucleotide diastereomers

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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