Characterization of DESI-FTICR mass spectrometry - from ECD to accurate mass tissue analysis.
Implementation of desorption electrospray ionization (DESI) technique on a 9.4 T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer is described. Desorption electrospray technique is capable of the direct investigation of natural samples without any need for sample preparation or chromatographic separation. Since the DESI mass spectra of natural samples are very complex owing to the lack of preseparation or cleanup, the ideal mass spectrometric analyzer for these applications is a high-resolution instrument such as FTICR mass spectrometer. DESI was implemented by constructing an electronically controlled source framework comprising six linear moving stages and one rotating stage. A three-dimensional linear stage was used to accommodate samples, while another 3D linear stage equipped with rotating stage was used as a spray mount. A modified electrosonic sprayer was used as a primary electrospray device. DESI-FTICR setup was characterized with regard to geometrical, electrical and flow conditions using deposited peptide samples in range of 1-100 pmol gross deposited amount on glass and polymer surfaces. Optimized conditions enabled the routine acquisition of DESI-MS spectra on the instrument at 130 000 resolution in the broadband mode and with comparable sensitivity to data reported in the literature. Since the main significance of DESI-FTICR MS is the combination of intact tissue analysis, the capabilities of the technique were demonstrated by analyzing murine liver samples. Presence of lysophospholipids in the liver tissue was tentatively associated with the lipid metabolism taking place in liver. DESI-FTICR is also a promising technique in the field of peptide analysis due to capability of top-down sequencing using electron capture dissociation. As a proof-of-principle experiment, a small synthetic polypeptide containing 36 amino acids was ionized using DESI and was sequenced in the FTICR by means of ECD (electron capture dissociation) fragmentation. Spectra gave almost full sequence information in agreement with the known amino acid sequence of the species.
Takats
Zoltan
Z
Semmelweis University, VIII. Ulloi ut 26, 1085 Budapest, Hungary.
Kobliha
Vaclav
V
Sevcik
Karel
K
Novak
Petr
P
Kruppa
Gary
G
Lemr
Karel
K
Havlicek
Vladimir
V
D000818:Animals; D016572:Cyclosporine / Q000737:chemistry; D004912:Erythrocytes / Q000737:chemistry; D005914:Globins / Q000032:analysis; D006418:Heme / Q000032:analysis; D006461:Hemolysis; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008246:Lysophospholipids / Q000032:analysis; D051379:Mice; D015203:Reproducibility of Results; D020539:Sequence Analysis, Protein; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
D008246:Lysophospholipids; D006418:Heme; D016572:Cyclosporine; D005914:Globins