N epsilon-(Carboxymethyl)lysine (CML) has been identified as a product of oxidation of fructoselysine (FL) in glycated (nonenzymatically glycosylated) proteins in vitro and has also been detected in human tissues and urine [Ahmed et al. (1986) J. Biol. Chem. 261, 4889-4894]. In this study, we compare the amounts of CML and FL in normal human lens proteins, aged 0-79 years, using specific and sensitive assays based on selected ion monitoring gas chromatography-mass spectrometry. Our results indicate that the lens content of FL increases significantly between infancy and about age 5 but that there is only a slight, statistically insignificant increase in FL between age 5 and 80 (mean +/- SD = 1.4 +/- 0.4 mmol of FL/mol of Lys). In contrast, the lens content of the oxidation product, CML, increased linearly with age, ranging from trace levels at infancy up to 8 mmol of CML/mol of lysine at age 79. The ratio of CML to FL also increased linearly from 0.5 to 5 mol of CML/mol of FL between age 1 and 79, respectively. These results indicate that CML, rather than FL, is the major product of glycation detectable in adult human lens protein. The age-dependent accumulation of CML in lens protein indicates that products of both glycation and oxidation accumulate in the lens with age, while the constant rate of accumulation of CML in lens with age argues against an age-dependent decline in free radical defense mechanisms in this tissue.

Reactive collisions of low energy (<100-eV) mass-selected ions are used to chemically modify fluorinated self-assembled monolayer surfaces comprised of alkanethiolate chains CF3(CF2)11(CH2)2S- bound to Au. Typical experiments were done by using 1-nA/cm(2) beams and submonolayer doses of reactant ions. Characterization of the modified surface was achieved by in situ chemical sputtering (60-eV Xe(+·)) and by independent high mass resolution time-of-flight-secondary ionization mass spectrometry (TOF-SIMS) (15-25-keV, Ga(+)) experiments. Treatment with Si(35)C1 4 (+·) produced a surface from which Xe(+) sputtering liberated CF2 (35)C1(+) ions, which suggested Cl-for-F halogen exchange at the surface. Isotopic labeling studies that used Si(35)Cl2 (37)Cl 2 (+·) ; and experiments with bromine-containing and iodine-containing projectiles, confirmed this reaction. High mass resolution TOF-SIMS spectra, as well as high spatial resolution images, provided further evidence as to the existence of halogen-exchanged species at the bombarded surface. Analogous Cl-for-F halogen substitution was observed in a model gas-phase reaction. The ion-surface reaction is suggested to proceed through an intermediate fluoronium ion in which the projectile is bonded to the target molecule. The most significant conclusion of the study is that selective chemical modification of monolayer surfaces can be achieved by using reactive ion beams, which lead to new covalent bonds at the surface and in the scattered ions.

A set of data pre-processing algorithms for peak detection and peak list alignment are reported for analysis of LC-MS based metabolomics data. For spectrum deconvolution, peak picking is achieved at selected ion chromatogram (XIC) level. To estimate and remove the noise in XICs, each XIC is first segmented into several peak groups based on the continuity of scan number, and the noise level is estimated by all the XIC signals, except the regions potentially with presence of metabolite ion peaks. After removing noise, the peaks of molecular ions are detected using both the first and the second derivatives, followed by an efficient exponentially modified Gaussian-based peak deconvolution method for peak fitting. A two-stage alignment algorithm is also developed, where the retention times of all peaks are first transferred into z-score domain and the peaks are aligned based on the measure of their mixture scores after retention time correction using a partial linear regression. Analysis of a set of spike-in LC-MS data from three groups of samples containing 16 metabolite standards mixed with metabolite extract from mouse livers, demonstrates that the developed data pre-processing methods performs better than two of the existing popular data analysis packages, MZmine2.6 and XCMS2, for peak picking, peak list alignment and quantification.

The ability of time-of-flight (TOF) mass spectrometry using an ultrahigh resolving power (R p of 100 000 at m/z 400) analyzer (HRT) to provide a proper platform for comprehensive petroleomic studies has been evaluated. The innovative HRT design for ultrahigh resolution using a "zig-zag" multi-reflecting analyzer with a folded flight path, efficient ion refocusing, and no substantial loss of ion transmission was tested. For comparison, samples were also analyzed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) using variable R p (from 100 000 at m/z 400 to 400 000 at m/z 400). Both HRT and FT-ICR MS data were processed and compared using software specifically designed to process either TOF or FT-ICR data for petroleomic studies. Owing to the increase in R p with m/z, the 100 000 at m/z 400 R p of the HRT is found to be, overall, comparable to that of a 200 000 at m/z 400 FT-ICR. This R p is sufficient to allow for proper resolution, correctly resolve most isobaric interferences, and accurately measure m/z values, leading to proper characterization of petroleum components via assignments of molecular formulas and classes. Results using electrospray ionization (ESI) focus on polar components, and their molecular formulas were properly translated to heteroatom classes, unsaturation levels measured via double bond equivalents (DBEs), and carbon number, which were displayed via classical geochemical plots. Data have shown that the compositional information provided by HRT is comparable to FT-ICR for most operational routine analyses. HRT operating with a R p of 100000 at m/z 400 was therefore found to offer a fast, accurate, sensitive, and cost-effective platform suitable for accurate petroleomic MS fingerprinting of crude oils and distillates.

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