Triversa™NanoMate

Triversa Nanomate or any other nanoflow electrospray ionization source... 

The benefits of nanoESI using the ESI Chip were recently extended to micro and analytical HPLC with the release of the TriVersa NanoMate system (Advion BioSystems) incorporating a post-column splitter that delivers 50 to 400 nL... 

Figure 3.9 Schematic of the TriVersa NanoMate enabling the coupling of LC columns with flow rates up to l.OmLmin-1 to the ESI Chip through use of a post-column splitter adjusted to deliver 200 to 400 nL min-1 of the postcolumn effluent to the ESI Chip while simultaneously collecting fractions into sample plates.

Another commercial system for proteomics research is the TriVersa NanoMate from Advion BioSciences Inc. It integrates LC-MS, fraction collection and chip-based infusion in one platform. Use of such chip systems is widespread in the literature . 

MS, MS/MS and MS experiments were performed with a LTQ-Orbitrap (Thermo Fisher Scientific Inc., San Jose, CA, USA) mass spectrometer equipped with a Triversa Nanomate (Advion Biosciences Inc., Ithaca, NY, USA). Compound solutions were infused with a nanospray chip. The mass spectrometer was first calibrated externally with a mixture containing caffeine, L-methionyl-arginyl-phenylalanyl-alanine (MRFA) and Ultramark 1621 in ACN, MeOH, HjO, acetic acid. Sub-ppm mass accuracy was finally achieved using an internal calibration (lock mass) in both MS and MS/MS mode. The resolution of Orbitrap MS was set to 100,000 (FWHM) at m/z 400. Electrospray ionization (ESI) was used. For MS/MS experiments, an isolation width of 1.5 Da was used. The normalized collision energy was set to the value when the precursor ion was exhausted. Helium was used as the collision gas. 

FIGURE 1.3 TriVersa NanoMate chip-based ESI system coupled to TLC see http // www.advion.com/products/triversa-nanomate/ for details on the technology, chip, and software. (Adapted from Advion Inc., Ithaca, NY.)... 

Online intact protein separation was the same as for the Q-TOF LC-MS (above) for consistent protein retention times across platforms. For LC-MS/MS the eluent flow was split to a flow rate of 350 nL/min via the TriVersa NanoMate (Advion BioSci-ences, Ithaca, NY) chip-based nanospray source and analyzed with a LTQ-Oibitrap XL (Thermo Fisher, San Jose, CA) mass spectrometer. The instrument was operated in a top-three data dependent mode, with both MS spectra and collision-induced dissociation (CID) MS/MS spectra acquired at 60,000 resolving power in the Oibi-trap. CID collision energy was operated at 15 %. Each MS spectrum was composed of three microscans, and each MS/MS spectrum was the average of 10 microscans. To facilitate the analysis of intact proteins, the instrument was operated with the HCD gas off and the delay before image current detection shortened to 5 ms. 

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