Specific Fragment Ions Research Articles

Distinguishing Endogenousd-Amino Acid-Containing Neuropeptides in Individual Neurons Using Tandem Mass Spectrometry

RNA-based protein synthesis produces L-amino acid-containing proteins and peptides. D-amino acid-containing peptides (DAACPs) can be generated from L-amino acid peptides via post-translational modification. In the nervous system, the conformational change of a single L-amino acid in a peptide to its D-form results in altered bioactivity, with some DAACPs having orders-of-magnitude enhanced efficacy. However, this modification is often overlooked when characterizing endogenous peptides. Here, with the use of matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF)/TOF mass spectrometry, neuropeptides that have the second residue isomerized to the D-isoform are distinguished from their L-epimers via differences in the relative amounts of specific fragment ions during tandem MS. With the appropriate fragment ions chosen, and in some cases with the use of metal adducts, epimer discrimination is optimized. Specifically, the cardioexcitatory peptide Asn-(D)Trp-Phe-amide (NdWFa) was assayed directly from neurons isolated from the sea slug Aplysia californica; the fraction of the peptide with the second residue (W) in the D- versus L-form was 90 ± 10%. We demonstrate that this approach is well suited for confirming DAACPs directly from cells and tissue, advancing our understanding of the l to d modification and the role it plays in cell-to-cell signaling.

LC Determination of Trace Biogenic Amines in Foods Samples with Fluorescence Detection and MS Identification

A pre-column derivatization method for the simple, sensitive determination of biogenic amines using 10-ethyl-acridine-3-sulfonyl chloride (EASC) as labeling reagent with fluorescence detection and mass spectrometry (MS) identification has been developed. After pre-column derivatization, the labeled biogenic amines were separated on a Hypersil BDS-C18 column by gradient elution. The derivatives showed an intense protonated molecular ion corresponding m/z [M + H]+ in positive-ion mode. The collision-induced dissociation of protonated molecular ion formed specific fragment ions at m/z 196.5, m/z 222.7, m/z 224.4 and m/z 272.5, m/z 286.2. Satisfactory linear responses were observed at the concentration range of 0.02–10 μmol L−1 with coefficients of >0.9993. Detection limits obtained by the analysis of a derivatized standard containing 0.2 pmol of each biogenic amine, were from 20.22 to 109.2 fmol (at a signal-to-noise ratio of 3). The relative standard deviations of retention times and peak areas for each biogenic amine were <0.96 and 3.22%, respectively. Recoveries except for PUT were in the range of 96.7–103.6% for chicken sausage and 95.8–104.6% for pork sausage The established method for the determination of biogenic amines except for PUT from real samples was satisfactory.

GC-MS Studies on Side Chain Regioisomers Related to Substituted Methylenedioxyphenethylamines: MDEA, MDMMA, and MBDB

Three regioisomeric 3,4-methylenedioxyphenethylamines having the same molecular weight and major mass spectral fragments of equal mass have been reported as drugs of abuse in forensic studies in recent years. These compounds are 3,4-methylenedioxy-N-ethylamphetamine (MDEA), 3,4-methylenedioxy-N,N-dimethylamphetamine (MDMMA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB). A series of seven additional side chain regioisomers have mass spectra essentially equivalent to the three controlled drug substances, all have molecular weight of 207 and major fragment ions in their electron ionization mass spectra at m/z 72 and 135/136. The trifluoroacetyl, pentafluoropropionyl, and heptafluorobutryl derivatives of the primary and secondary regioisomeric amines were evaluated in GC-MS studies. The mass spectra for these derivatives were significantly individualized, and the resulting unique fragment ions allowed for specific side chain identification. The trifluoroacetyl and heptafluorobutryl derivatives provided more specific fragment ions for molecular individualization among these regioisomeric substances. These perfluoroacyl derivatives showed reasonable resolution on the polar stationary phase Rtx-200.

Determination of Free Fatty Acids in Tibet Folk Medicine Potentilla anserina L. Using a New Labeling Reagent by LC with Fluorescence Detection and Identification with Online Atmospheric Chemical Ionization-MS Identification

A sensitive method for the determination of free fatty acids using 1,2-benzo-carbazole-9-ethyl-p-toluenesulfonate (BCETS) as tagging reagent with fluorescence detection has been developed. BCETS could easily and quickly label fatty acids in the presence of the K(2)CO(3) catalyst at 80 A degrees C for 30 min in N,N-dimethylformamide solvent. In this study, fatty acids from the extracted Potentilla anserina L. plant sample were sensitively determined. The corresponding derivatives were separated on a reversed-phase Eclipse XDB-C(8) column by LC in conjunction with gradient elution. The identification was carried out by post-column APCI-MS in positive-ion detection mode. BCETS-fatty acid derivatives gave an intense molecular ion peak at m/z [M+H](+), the collision-induced dissociation spectra of m/z [M+H](+) produced the specific fragment ions at m/z [M'+CH(2)CH(2)](+), m/z 216.6 and m/z [MH-H(2)O](+) (here, M': corresponding molecular mass of the fatty acids). The fluorescence excitation and emission wavelengths of the derivatives were at lambda (ex) 279 nm and lambda (em) 380 nm, respectively. Linear correlation coefficients for all fatty acid derivatives are more than 0.9994. Detection limits, at a signal-to-noise ratio of 3:1, are 10.79-34.19 fmol for the labeled fatty acids.

HPLC-FLUORIMETRIC METHOD FOR ANALYSIS OF FREE FATTY ACIDS IN STELLERA CHAMAEJASMA L

A sensitive HPLC method for the determination of fatty acids using 2-(2-(pyren-1-yl)-1H-benzo[d]imidazol-1-yl) ethyl-p-toluenesulfonate (PBIOTs) as a novel labeling reagent with fluorescence detection has been developed. PBIOTs could easily and quickly label fatty acids in the presence of the K2CO3 catalyst at 90°C for 30 min in N,N-dimethylformamide solvent. Fatty acids derivatives were separated on a reversed phase Eclipse XDB-C8 column by HPLC in conjunction with gradient elution. The corresponding derivatives were identified by post-column APCI-MS in positive ion detection mode. PBIOTs-fatty acid derivatives gave an intense molecular ion peak at m/z [M + H]+; with MS/MS analysis, the collision induced dissociation spectra of m/z [M + H]+ produced the specific fragment ions at m/z [M + H–319]+ and m/z 319.0 (here, m/z 319 is the core structural moiety of the PBIOTs molecule). The fluorescence excitation and emission wavelengths of the derivatives were λex = 350 nm and λem = 402 nm, respectively. Linear correlation coefficients for all fatty acid derivatives were ≥0.9985. Detection limits for the labeled fatty acids, at a signal-to-noise ratio of 3:1, were in the range of 10.32– 44.28 fmol. From the extracts of Stellera chamaejasma L, free fatty acids were sensitively determined.

2-(5-Benzoacridine)ethyl-p-toluenesulfonate as sensitive reagent for the determination of bile acids by HPLC with fluorescence detection and online atmospheric chemical ionization-mass spectrometric identification

2-(5-Benzoacridine)ethyl-p-toluenesulfonate (BAETS), a dual-sensitive probe, was reacted with bile acids in the presence of K(2)CO(3) catalyst in dimethyl sulfoxide (DMSO) solvent to give BAETS-bile acid derivatives. Derivatives exhibited intense fluorescence (FL) with an excitation maximum at lambda(ex) 270 nm and an emission maximum at lambda(em) 510 nm. MS analysis using APCI-MS indicated that derivatives had excellent APCI-MS ionizability with percentage ionization delta values changing from 0 to 88.83% in aqueous acetonitrile and from 0 to 89.15% in aqueous methanol. The collision induced dissociation spectra of m/z [M + H](+) contained specific fragment ions at m/z [M + H-H(2)O](+), [M + H-2H(2)O](+), [M + H-3H(2)O](+), 347.3, and 290.1. Repeatability was good for LC separation of BAETS-bile acid derivatives with aqueous acetonitrile as mobile phase. The relative standard deviations (RSDs) of retention time and peak area at 6.6 nmol mL(-1) levels with fluorescence detection (FL) were from 0.045 to 0.072% and from 2.16 to 2.73%, respectively. Excellent linear responses were observed, with regression coefficients >0.9995. The FL detection limits (S/N = 3) were in the range of 18.0-36.1 fmol. The online APCI-MS detection limits are in the range of 500-790 fmol (at a signal-to-noise ratio of 3).

Minimally permutated peptide analogs as internal standards for relative and absolute quantification of peptides and proteins

A novel type of isobaric internal peptide standard for quantitative proteomics is described. The standard is a synthetic peptide derived from the target peptide by positional permutation of two amino acids. This type of internal standard is denominated minimally permutated peptide analog (MIPA). MIPA can be differentiated from their target analytes by LC-MS due to individual retention times and/or by MS/MS due to specific fragment ions. Both quantification methods are demonstrated using peptide mixtures of low and high complexity.

Residual Agar Determination in Bacterial Spores by Electrospray Ionization Mass Spectrometry

Presented here is an analytical method to detect residual agar from a bacterial spore sample as an indication of culturing on an agar plate. This method is based on the resolubilization of agar polysaccharide from a bacterial spore sample, enzymatic digestion, followed by electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis for detection of a specific agar fragment ion. A range of Bacillus species and strains were selected to demonstrate the effectiveness of this approach. The characteristic agar fragment ion was detected in the spores grown on agar that were washed from 1 to 5 times, irradiated or nonirradiated, and not in the spores grown in broth. A sample containing approximately 10(8) spores is currently needed for confident detection of residual agar from culture on agar plates in the presence of bacterial spores with a limit of detection of approximately 1 ppm agar spiked into a broth-grown spore sample. The results of a proficiency test with 42 blinded samples are presented demonstrating the utility of this method with no false positives and only three false negatives for samples that were below the detection level of the method as documented.

Positive and negative tandem mass spectrometric fingerprints of lipids from the halophilic Archaea Haloarcula marismortui

Lipids from the extremely halophilic Archaea, Haloarcula marismortui, contain abundant phytanyl diether phospholipids, namely archaetidic acid (AA), archaetidylglycerol (AG), archaetidylglycerosulfate (AGS), with mainly archaetidylglycerophosphate methyl ester (AGP-Me). These were accompanied by a triglycosyl archaeol (TGA), lacking characteristic sulfate groups. Tandem-mass spectrometry was employed to provide fingerprints for identifying these known lipids, as well as small amounts of unsaturated phospholipids. These contained 3 and 6 double bonds in their archaeol moiety, suggested by negative tandem-MS of intact phospholipids, as indicated by differences between their pseudo-molecular ion and specific fragment ions designated as pi(2). The core ether lipids were confirmed by electrospray ionization mass spectrometry (ESI-MS) as 2,3-di-O-phytanyl-sn-glycerol (C20, C20), which gave rise to a precursor-ion at m/z 660 [M+Li](+), and its fragment ion at m/z 379 [M+Li](+), consistent with mono-O-phytanyl-glycerol. Furthermore, lithiated ions at m/z 654 (MS(1)), 379 (MS(2)) and m/z 648 (MS(1)), 373 (MS(2)), combined with (1)H/(13)C NMR chemical shifts at delta 5.31-121.6 (C2/2'-H2/2'), 5.08-124.9 (C6/6'-H6/6') and 5.10-126.0 (10/10'-H10/10') confirmed the presence of unsaturated homologs of archaeol. We carried out a comprehensive study on the lipids present in cells of H. marismortui. We used positive and negative ESI-MS with tandem-MS, which served as a fingerprint analysis for identifying the majority of component lipids.

Improving the selectivity of non-polar alkanes, alkenes, and aromatic hydrocarbon compounds by the application of acetonitrile as a Cl reagent

Abstract The selective enhancement of membrane introduction mass spectrometry for non-polar alkanes, alkenes, and aromatic hydrocarbon compounds by the application of acetonitrile as a chemical ionization reagent was investigated. Acetonitrile Cl is able to produce specific fragment ions for many of the compounds test and this can be used to identify and quantify the parent neutrals. This method provided relatively high detection limits of the test compounds. This method could potentially be useful for analytical applications such as the detection of non-polar hydrocarbons for environmental studies if CH3CN Cl/MIMS is coupled with a preconcentration method.

Integrative Normalization and Comparative Analysis for Metabolic Fingerprinting by Comprehensive Two-Dimensional Gas Chromatography−Time-of-Flight Mass Spectrometry

Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC x GC-TOF-MS) was applied to the comparative metabolic fingerprinting of a wild-type versus a double mutant strain of Escherichia coli lacking the transhydrogenases UdhA and PntAB. Using peak lists generated with the Leco ChromaTOF software as input, we developed retention time correction and data alignment tools (INCA). The accuracy of peak alignment and detection of 1.1- to 4-fold changes in metabolite concentration was validated by a spike-in experiment with 20 standard compounds. A list of 48 significant features that differentiated the two E. coli strains was obtained with an estimated false discovery rate (FDR) of <0.05. A total of 27 metabolites, mainly from the citrate cycle, were identified. That the signal intensity of the m/z 73 trace of the trimethylsilyl (TMS) group reflected true differences in metabolite abundance was confirmed by quantification of pyruvate, fumarate, malate, succinate, alpha-ketoglutarate, citrate, cis-aconitate, myo-inositol, and glucose-6-phosphate using compound specific fragment ions and stable isotope labeled standards. Relative standard deviations for metabolite extraction and GC x GC-TOF-MS analysis of those analytes ranged from 13.2 to 26.3% for the universal m/z 73 trace and 7.4 to 24.5% for the analyte specific fragment ion trace.

Alkylation of complementary ribonucleotides by 1,2‐dodecyl‐epoxide in a micellar environment: a liquid chromatography—electrospray ionization—sequential mass spectrometry investigation

Alkylation of a pair of complementary ribonucleotides, adenosine monophosphate (AMP) and uridine monophosphate (UMP), was accomplished by 1,2-dodecyl-epoxide (DE) in a oil-in-water microemulsion based on the cationic surfactant Cetyl-trimethyl-ammonium-bromide, providing a suitable catalytic interface for the reagents. Several, often isomeric, alkylation products, bearing one or two hydroxy-dodecyl moieties on their structures, were identified in the reaction mixtures by high-performance liquid chromatography coupled to electrospray ionization ion trap mass spectrometry. In particular, mass spectrometry (MS)/MS spectra, implemented by extracted ion chromatograms obtained for peculiar MS/MS product ions, indicated alkylation to occur on uracil and on uracil/phosphate OH groups in singly and doubly alkylated UMP, respectively. Adenine NH2 group and phosphate or ribose OH groups were found to be involved as such (single alkylation) or in combination, in the case of alkylated derivatives of AMP. The reaction of both endocyclic N and C=O groups (tautomerized to C-OH groups) of uracil and the predominance of nucleophilic attack to the more accessible carbon of the DE epoxydic bridge (the only exception being the reaction by the NH2 group of adenine) were inferred from MS3 spectra with the help of extracted ion chromatograms for specific fragment ions, after their structural characterization. Interestingly, alkylation on one of the uracil C=O groups and, partially, on the adenine NH2 group, both potentially involved in AMP/UMP base pairing in the micellar environment, were found to be hindered when both ribonucleotides were present in the reaction mixtures.

Structural determination by negative-ion MALDI-QIT-TOFMSn after pyrene derivatization of variously fucosylated oligosaccharides with branched decaose cores from human milk

We prepared neutral oligosaccharide fraction from milk of a woman (blood type A, Le(b+)) by anion-exchange column chromatography after the removal of lipids and proteins. Further fractionation was performed by means of Aleuria aurantia lectin-Sepharose column chromatography and reverse-phase HPLC after labeling with a pyrene derivative. This pyrene labeling allowed identification by negative-MALDI-TOFMS(n) analysis of 22 oligosaccharides with decaose cores, among which 21 had novel structures. Negative ions could not be produced from neutral oligosaccharides without labeling on MALDI. Mono-, di-, tri-, and tetrafucosylated decaose fractions contained three, nine, six, and four isomers, respectively. Our method enables easy determination of fucosylated structures on the N-acetyllactosamine branches of these isomers. On negative-MS(n) the fragment ions included several A and D ions, from which fucosylation on the branches could be elucidated. Other characteristic ions were also detected. Y-type cleavage at the reducing side of -3GlcNAc indicated the occurrence of type 1 chain. Specific fragment ions were produced from H, Le(a), and Le(x) antigens. Linkage-specific exoglycosidase digestion confirmed the structures. The results indicate that the diversity of the oligosaccharides is due to combinations of type 1 H, Le(a), Le(x), and Le(b)/Le(y) on branched decaose cores. In typical oligosaccharides, 6-branches always consist of type 2 chain, while 3-branches, such as beta and gamma chains, are fucosylated type 1 chains. From the viewpoint of biosynthesis, the presence of fucosylation and type 1 chain may halt elongation of the N-acetyllactosamine and promote formation of branched structures.

Identification of autoxidation oligomers of flavan‐3‐ols in model solutions by HPLC‐MS/MS

Autoxidation of flavan-3-ols was carried out in aqueous/methanol model solutions under mildly acidic conditions (pH 6.0), and these autoxidation products were analyzed by using high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS). The results showed that (+)-catechins and (-)-epicatechins generated autoxidation reaction with each other to form a series of oligomers that had the same [M - H](-) molecular ions (MS(1)) as those of natural procyanidins, but had completely different fragment ions (MS(2)). According to MS/MS analysis, the major fragments of these oligomers were derived not only from the retro-Diels-Alder (RDA) dissociations on the C-rings of the flavan-3-ol units, but also from the quinone-methide (QM) cleavage of the interflavan linkages (IFL), and thus they were identified as B-type dehydrodicatechins, B-type dehydrotricatechins and A-type dehydrotricatechins, respectively. The potential structures of their [M - H](-) molecular ions and partial fragment ions were deduced on the basis of the MS/MS characterization and the oxidation of flavan-3-ols in previous reports. Some specific fragment ions were found to be very useful for identifying the autoxidation oligomers (the B-type dehydrodicatechins at m/z 393, the B-type dehydrotricatechins at m/z 681 and the A-type dehydrotricatechins at m/z 725).

Multiple ionization ofC60by fastSiq+ions

Multiple ionization of ${\mathrm{C}}_{60}$ by ${\mathrm{Si}}^{q+}$ ions $(q=1--3)$ has been studied at incident energies of 0.5, 0.8, 2.0, and $6.0\phantom{\rule{0.3em}{0ex}}\mathrm{MeV}$ covering a velocity range of $v=0.85--2.93$ in atomic units. Fragment ions and secondary electrons were measured in coincidence with charge-selected outgoing projectile ions, enabling us to determine the charge state of prefragmented parent ions $\mathrm{C}_{60}{}^{r+}$. Charge state $(r)$ distributions correlated with the smallest fragment ions of $\mathrm{C}_{m}{}^{+}\phantom{\rule{0.3em}{0ex}}(m\ensuremath{\leqslant}3)$ were found to vary rather strongly depending on $v$ and $q$, whereas the $r$ distributions were less sensitive for medium sized ions of $m\ensuremath{\geqslant}4$. These results reflect fairly well the difference of the amount of inelastic energy deposition estimated from the local density approximation. Experimental $r$ distributions correlated with specific fragment ions $\mathrm{C}_{m}{}^{+}\phantom{\rule{0.3em}{0ex}}(m\ensuremath{\leqslant}5)$ were reproduced almost perfectly by our statistical energy deposition model. From our model calculations we also obtained the most probable values of the ionization energy associated with the production of individual fragment ions.

A δ-Catenin Signaling Pathway Leading to Dendritic Protrusions

Delta-catenin is a synaptic adherens junction protein pivotally positioned to serve as a signaling sensor and integrator. Expression of delta-catenin induces filopodia-like protrusions in neurons. Here we show that the small GTPases of the Rho family act coordinately as downstream effectors of delta-catenin. A dominant negative Rac prevented delta-catenin-induced protrusions, and Cdc42 activity was dramatically increased by delta-catenin expression. A kinase dead LIMK (LIM kinase) and a mutant Cofilin also prevented delta-catenin-induced protrusions. To link the effects of delta-catenin to a physiological pathway, we noted that (S)-3,5-dihydroxyphenylglycine (DHPG) activation of metabotropic glutamate receptors induced dendritic protrusions that are very similar to those induced by delta-catenin. Furthermore, delta-catenin RNA-mediated interference can block the induction of dendritic protrusions by DHPG. Interestingly, DHPG dissociated PSD-95 and N-cadherin from the delta-catenin complex, increased the association of delta-catenin with Cortactin, and induced the phosphorylation of delta-catenin within the sites that bind to these protein partners.

Analysis of aldehydes via headspace SPME with on‐fiber derivatization to their O‐(2,3,4,5,6‐pentafluorobenzyl)oxime derivatives and comprehensive 2D‐GC‐MS

A method for the analysis of the homologous series of alkanals, (E)-2-alkenals, and (E,E)-2,4-alkadienals is described utilizing a headspace solid-phase microextraction (HS-SPME) step and on-fiber derivatization with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) hydrochloride. Oxime derivatives formed on the fiber are desorbed in the gas chromatographic injector and analyzed by comprehensive 2-D GC coupled to quadrupole MS (GC x GC-qMS). Selecting specific fragment ions within the electron impact mass spectra of the oxime derivatives provides a suitable method for the target analysis of these aldehyde classes, which furthermore benefits from the increased separation efficiency by GC x GC. The analysis of higher molecular weight aldehydes is described in wine and grape seed oil as examples. Quantification of the aldehydes utilizes a stable isotope dilution analysis (SIDA) assay with octan-d(16)-al as isotopomeric internal standard. Besides the selectivity and sensitivity of aldehyde analysis using PFBHA derivatives, critical aspects on background level contamination and repeatability of the sample preparation method are discussed. Optimization of GC x GC-qMS parameters allowed a considerable saving of the cryogenic medium, involving additional (unmodulated) conditioning runs, rendering the method more amenable to routine analysis.

LC-DAD-ESI-MS Characterization of Carbohydrates Using a New Labeling Reagent

A novel labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) coupling to liquid chromatography with electrospray ionization mass spectrometry for the detection of carbohydrates from the derivatized rape bee pollen samples is reported. Carbohydrates are derivatized to their bis-NMP-labeled derivatives. Derivatives showed an intense protonated molecular ion at m/z [M+H]+ in positive-ion detection mode. The mass-to-charge ratios of characteristic fragment ions at m/z 473.0 could be used for the accurately qualitative analysis of carbohydrates. This characteristic fragment ion is from the cleavage of C2–C3 bond in carbohydrate chain giving the specific fragment ions at m/z [MH-C m H2m+1O m -H2O]+ for pentose, hexose and glyceraldehydes and at m/z [MH-C m H2m-1O m+1-H2O]+ for alduronic acids such as galacturonic acid and glucuronic acid (m = n − 2, n is carbon number of carbohydrate). No interferences for all aliphatic and aromatic aldehydes presented in natural environmental samples were observed due to the highly specific parent mass-to-charge ratio and the characteristic fragment ions. The method, in conjunction with a gradient elution, offered a baseline resolution of carbohydrate derivatives on a reversed-phase Hypersil ODS-2 column. The carbohydrates such as mannose, galacturonic acid, glucuronic acid, rhamnose, glucose, galactose, xylose, arabinose and fucose can successfully be detected.

Analysis of Carbohydrates in a Tibetan Medicine Using New Labeling Reagent, 1-(2-Naphthyl)-3-Methyl-5-Pyrazolone, by HPLC with DAD Detection and ESI-MS Identification

A new labeling reagent, 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP), coupling with liquid chromatography (LC) with electrospray ionization mass spectrometry (ESI-MS) for the detection of carbohydrates from a famous Tibetan medicine is reported. Carbohydrates were derivatized to their bis-NMP-labeled derivatives. The method, in conjunction with a gradient elution, offered a baseline resolution of carbohydrate derivatives on a reversed phase Hypersil ODS-2 column. The carbohydrates such as mannose, galacturonic acid, glucuronic acid, rhamnose, glucose, galactose, xylose, arabinose, and fucose could be successfully detected by UV and ESI-MS. Derivatives showed intense protonated molecular ion at m/z [M + H]+ in positive ion mode. The mass to charge ratios of characteristic fragment ions at m/z 473.0 could be used for the accurately qualitative identification of carbohydrates; this characteristic fragment ion was from the cleavage of C2-C3 bond in the carbohydrate chain giving the specific fragment ions at m/z [MH–C m H2m+1O m –H2O]+ for pentose, hexose, and glyceraldehydes, and at m/z [MH–C m H2m−1O m+1–H2O]+ for alduronic acids, such as galacturonic acid and glucuronic acid (m = n − 2, n is carbon atom number of carbohydrate). Compared with the traditional 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent, currently synthesized NMP show the advantage of higher sensitivity to carbohydrate compounds with UV and ESI-MS detection.

Identification of Monosialylated N-glycoforms in the CDG Urinome by Ion Mobility Tandem Mass Spectrometry: The Potential for Clinical Applications

Abstract Introduction A novel approach of ion mobility tandem mass spectrometry (IMS-MS/MS) is applied to analysis of human glycourinome to obtain carbohydrate pattern data of congenital disorders of glycosylation patient. Overlapping of the complex carbohydrate mass range landscape has been highly reduced upon IMS-MS procedure, allowing more efficient identification by mapping and sequencing of glycan precursor ions, following their separation by mobility, according to difference in drift time through the traveling wave IMS cell. Intact and truncated N- and O-glycan structures modified by sialylation and fucosylation were identified according to their drift time separated molecular ions and submitted to fragmentation in a narrow mass window. IMS CID MS/MS Analysis The fragmentation spectra generated from the IMS separated precursor ions contain series of fragment ions maintaining the same mobility as their parent ions, and the assignment accuracy can be significantly enhanced. Conclusion According to the specific fragment ion patterns, carbohydrate epitopes described to be involved in pathological processes were assigned. A high potential of this glycomics-based strategy for clinical applications can be presented.