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81.
Schiopu C Vukelić Z Capitan F Kalanj-Bognar S Sisu E Zamfir AD 《Electrophoresis》2012,33(12):1778-1786
A strategy combining high-performance thin layer chromatography (HPTLC), laser densitometry, and fully automated chip-based nanoelectrospray (nanoESIchip) performed on a NanoMate robot coupled to QTOF-MS was developed, optimized, and for the first time applied for mapping and structural identification of gangliosides (GGs) extracted and purified from a human angioblastic meningioma specimen. While HPTLC pattern indicated only seven fractions migrating as GM3, GM2, GM1, GD3, GD1a (nLD1, LD1), GD1b, GT1b, and possibly GD2, due to the high sensitivity, mass accuracy, and ability to ionize minor species in complex mixtures, nanoESIchip-QTOF MS was able to discover significantly more GG species than ever reported in meningioma. Thirty-four distinct glycosphingolipid components of which five asialo, one GM4, nine GM3, two GM2, two GD3, nine GM1, and six GD1 differing in their ceramide compositions were identified. All structures presented long-chain bases with 18 carbon atoms, while the length of the fatty acid was found to vary from C11 to C25. MS screening results indicated also that the diversity of the expressed GM1 structures is higher than expected in view of the low proportions evidenced by densitometric quantification. Simultaneous fragmentation of meningioma-associated GM1 (d18:1/24:1) and GM1 (d18:1/24:0) by MS/MS using CID confirmed the postulated structures of the ceramide moieties and provided data on the glycan core, which document that for each of the GM1 (d18:1/24:1) and GM1 (d18:1/24:0) forms both GM1a and GM1b isomers are expressed in the investigated meningioma tissue. 相似文献
82.
Attila Gáspár István Bácsi Erika F. Garcia Mihály Braun Frank A. Gomez 《Analytical and bioanalytical chemistry》2009,395(2):473-478
The goal of this work was to increase the sensitivity of a UV–Vis spectrophotometer by decreasing the background noise and
lengthening the optical path. A microphotometer has been modified to precisely select very small parts of a microfluidic channel
pattern of a chip and to measure light absorbance on a magnified area of the selected part of the channel. The viability of
combining a projection microscope and a spectrophotometer for external absorbance measurements on disposable PDMS chips was
studied. Besides the external direct detection above a microfluidic channel, the optical pathlength was lengthened by detecting
in the region of the perpendicular exit port. Increasing the cross-sectional area of the zone of irradiation improved the
signal-to-noise ratio and the limits of detection (LOD). 相似文献
83.
Luciano Callipo Patrizia Foglia Riccardo Gubbiotti Roberto Samperi Aldo Laganà 《Analytical and bioanalytical chemistry》2009,394(3):811-820
A method for carbonic anhydrase II (CA II) absolute quantification in human serum is presented. This method is based on high-performance
liquid chromatography (HPLC)-Chip microfluidic device incorporating a nanoelectrospray source interfaced to a triple quadrupole
mass spectrometer. The fraction containing CA II was isolated by preparative reversed-phase HPLC, and peptides obtained from
the tryptic digest of the protein mixture were separated by the HPLC-Chip system. The multiple-reaction monitoring acquisition
mode of a selected suitable CA II peptide and peptide internal standard allowed the selective and sensitive determination
of a CA II. Absolute recovery of the method was 52 ± 12%, while analytical recovery was 81 ± 10%. For the eight samples analyzed,
the matrix effect was found to be only −14 ± 6%. A comparison among three regression lines type which were obtained by external
calibration, matrix-matched calibration, and standard addition method, respectively, demonstrated that the first one is adequate
in obtaining good accuracy and precision. Method quantification limit for CA II in serum was estimated to be 2 fmol/mL. CA
II mean concentration in sera from eight healthy subjects was found to be 56 pmol/mL (relative standard deviation 24%). 相似文献
84.
Hans‐Achim Wagenknecht 《Chemie in Unserer Zeit》2002,36(5):318-330
DNA charge transfer chemistry has been subject of considerable interest with consequences in the formation of oxidative damage to the DNA which can result in mutagenesis or carcinogenesis. In this article, important examples of spectroscopical and biochemical assays are compared and discussed in terms of the effiencies, rates, and mechanisms. Coupled with the demonstration that such charge transfer can be modulated both negatively and positively by DNA‐binding proteins, these observations therefore suggest the intriguing possibility that DNA‐mediated charge transfer chemistry is biological relevant and may play a role in cellular processes. Additionally, charge transfer chemistry plays a growing role in the recent development of DNA chips detecting mutations or lesions of nucleic acids. 相似文献