Protein analysis for nitrogen generator applications

The Thermo Scientific™ Orbitrap Eclipse™ mass spectrometer is designed to solve analytical challenges in biological research and easily meet the challenges of proteomics, biopharmaceutical and advanced protein structure analysis research.

Recently, we visited a Shanghai-based scientific research institution conducting protein analysis to assess the performance and operation of PEAK Scientific’s LC-MS grade nitrogen generators in combination with a Thermo Scientific™ Orbitrap Eclipse™ mass spectrometer.

When performing mass spectrometry analysis, it is necessary to separate the pre-treated sample in liquid phase, and then to convert it from a liquid state to a gaseous state as it enters the ion source. To achieve this, the ion source needs to be fed with nitrogen with a specific flow, pressure and purity greater than 95%. As a gas for atomization and drying in this process, it is very important that the generator can provide a stable and reliable gas quality.

Thermo Scientific™ Orbitrap Eclipse™ Mass Spectrometer

The Genius NM32LA nitrogen generator used by the customer is a tried and tested PEAK product. It uses membrane separation technology and is equipped with a built-in air compressor, which can generate analytical-grade nitrogen on site and on demand.

PEAK Genius NM32LA Nitrogen Generator

01 Materials and Methods

Device Information:

Mass Spec Model: Thermo Scientific™ Orbitrap Eclipse™ Mass Spectrometer, FAIMS Pro Interface

Generator Model: PEAK Genius NM32LA Nitrogen Generator

Sample Information:

Sample: 293T cells, prepared in 1% formic acid water

Blank: 1% formic acid water

Instrument method:

Chromatographic column: reversed phase C18; chromatographic column, 30cm*75um

Mobile phase: A phase: formic acid water; B phase: formic acid acetonitrile

Analysis time: 60min

02 Test results

Figure 1. TIC total ion chromatogram, BPC ion chromatogram, and secondary ion chromatogram of solvent blank

Figure 2. Solvent Blank Baseline Zoom

Figure 3. Sample test spectrum TIC total ion chromatogram, BPC ion chromatogram, secondary ion chromatogram

03 Results Discussion

The nitrogen gas introduced into the ion source part should not react with the sample and should be free from impurities since this can affect analysis and increase the maintenance frequency of the instrument. The flow, purity and quality of nitrogen, as well as the stability and efficiency of the gas supply system itself, can therefore affect the overall operation of mass spectrometry.

The results shown in the TIC total ion chromatogram, BPC ion chromatogram, and secondary ion chromatogram of the solvent blank and the sample show that the spectrum baseline is stable and no interference from impurity peaks was observed. The results clearly demonstrate that the Genius NM32LA nitrogen generator provides the necessary nitrogen flow, pressure and purity for this analysis. The generator can continuously provide gas that meets the requirements for the MS ion source and fully supports this analysis.

As a gas generator manufacturer specializing in the laboratory field, PEAK Scientific provides stable and reliable gas generator products, which not only provides a convenient source of gas but also ensures that mass spectrometers operate efficiently and consistently. Choosing a suitable nitrogen generator can not only save money, but also ensures consistent gas purity and quality improving the efficiency of lab workflows for LC-MS analysis.

About Ed Connor
Ed Connor joined PEAK Scientific in February 2013 as a GC product specialist and now functions as a Product Manager.

He has been working on a number of collaborative projects with PEAK customers and the major instrument manufacturers worldwide.

The main focus of these collaborations has been to look at conversion from helium to hydrogen or nitrogen carrier gas for GC applications but he has also worked on ELSD nitrogen gas solutions and nitrogen generators for LC-MS instruments.

Prior to joining PEAK, Ed completed his Dr.Sc. at ETH Zurich in 2007 using GC-MS to look at herbivore induced plant volatiles and their interaction with beneficial insects.

He then joined the University of Zurich where his work focused primarily on floral volatile analysis using a variety of volatile collection methods, GC-MS and GC-FID.