PubMed · 5634641
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J S Annino. 1968. A sequential sampling system for spectrophotometers.. https://pubmed.ncbi.nlm.nih.gov/5634641/
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Spectra of highly acidic oligosaccharides obtained by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) can be greatly enhanced in quality by coating the MALDI probe surface with a film consisting of a perfluorosulfonated ionomer (Nafion) prior to the addition of the sample-matrix mixture. For comparison, three mixtures containing highly acidic oligo- and polysaccharides derived from partial acidic hydrolysis of alginate, pectin, or carboxymethyl cellulose (CMC) were analyzed by employing probes with an uncoated gold surface or a surface coated with a Nafion or nitrocellulose film. The negative ion-mode MALDI spectra of the oligouronates (oligomers containing mannuronic/guluronic and galacturonic acid residues) obtained using uncoated or nitrocellulose-coated probes consisted of a series of broad, multiple peaks. These multiple peaks were assigned to the molecular ions of the nondissociated [M - H]- and partially sodiated [MnNa - H]-, where n = 1, 2, or 3, oligomers. In contrast, the corresponding spectra obtained with Nafion-coated probes contained only a single series of sharp peaks originating from the molecular ions ([M - H]-) of nondissociated oligomers exhibiting chain lengths of as many as approximately 15 uronic acid residues. The Nafion coating was apparently capable of removing the sodium counterions remaining in the deposit of the sample-matrix mixture on the probe, thereby greatly enhancing the signal-to-noise ratios of the peaks in the spectra. In a similar manner, higher quality spectra could also be obtained by using Nafion-coated probes for analysis of the oligouronates and CMC oligomers by positive ion-mode MALDI-MS.
The use of an active Nafion substrate is shown to enhance the performance of MALDI MS. The use of a Nafion substrate with certain matrices can significantly enhance the signals obtained over those observed with a stainless steel probe substrate. Analytes can often be observed with the use of the Nafion substrate that cannot be easily observed with the standard MALDI procedure, and usually a much wider range of peaks can be observed using MALDI from the Nafion substrate than with any single matrix on a stainless steel substrate. This enhancement of signal from the Nafion substrate is observed only with the sequential deposition of the sample and the matrix onto the Nafion film. If the analyte and matrix are premixed, then the effect is not observed. The use of the Nafion substrate has been shown to be particularly effective in analyzing real biological mixtures without prepurification. This has been demonstrated for various samples including the analysis of the products of chemical digests of proteins, protein profiling in milk and egg white samples, cell lysate analysis, and oligonucleotide detection.
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