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Dive into the research topics where Alina D. Zamfir is active.

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Featured researches published by Alina D. Zamfir.


Analytical Biochemistry | 2008

Coupling of fully automated chip-based electrospray ionization to high-capacity ion trap mass spectrometer for ganglioside analysis

Reinaldo Almeida; Cristina Mosoarca; Marius Chirita; Valentina Udrescu; Nicolae Dinca; Željka Vukelić; Mark Allen; Alina D. Zamfir

NanoMate robot was coupled to a high-capacity ion trap (HCT) mass spectrometer to create a system merging automatic chip-based electrospray ionization (ESI) infusion, ultrafast ion detection, and multistage sequencing at superior sensitivity. The interface between the NanoMate and HCT mass spectrometer consists of an in-laboratory constructed mounting device that allows adjustment of the robot position with respect to the mass spectrometer inlet. The coupling was optimized for ganglioside (GG) high-throughput analysis in the negative ion mode and was implemented in clinical glycolipidomics for identification and structural characterization of anencephaly-associated species. By NanoMate HCT mass spectrometry (MS), data corroborating significant differences in GG expression in anencephalic versus age-matched normal brain tissue were collected. The feasibility of chip-based nanoESI HCT multistage collision-induced dissociation (CID MS(n)) for polysialylated GG fragmentation and isomer discrimination was tested on a GT1 (d18:1/18:0) anencephaly-associated structure. MS(2)-MS(4) obtained by accumulating scans at variable fragmentation amplitudes gave rise to the first fragmentation patterns from which the presence of GT1b structural isomer could be determined unequivocally without the need for supplementary investigation by any other analytical or biochemical methods.


Electrophoresis | 2001

Glycoscreening by on-line sheathless capillary electrophoresis/electrospray ionization-quadrupole time of flight-tandem mass spectrometry

Alina D. Zamfir; Jasna Peter-Katalinić

An analytical approach based on sheathless on‐line coupling of capillary electrophoresis (CE) and electrospray ionization (ESI) quadrupole time‐of‐flight (QTOF) mass spectrometry (MS) has been developed for providing new insight into the characterization of carbohydrate mixtures. The home‐built sheathless CE/ESI microspray interface consisting of an one‐piece CE column has been optimized and applied for the screening of a complex mixture of O‐glycosylated peptides obtained and previously purified from urine of patients suffering from N‐acetylhexosaminidase deficiency. The separation efficiency and the resolution obtained in CE/UV experiments could be reproduced with high sensitivity in on‐line CE/MS runs under mild ES‐Inegative ion mode conditions, due to the compatibility of the reconsidered CE‐QTOF/MS operating parameters, microspray tip performance and the use of MS friendly volatile CE buffer system as ammonium acetate/ammonium hydroxide. By employing the high speed automated “on‐the‐fly” MS‐MS/MS switching abilities of QTOF/MS instrument, the multicomponent sample could be separated and subsequently submitted to both MS and low‐energy collision‐induced dissociation of selected precursor ions. Using the on‐line CE‐ESI‐MS/MS coupling, glycosylated sialylated peptides undetectable in complex mixtures by a direct ESI/MS analysis could be identified.


Electrophoresis | 2002

A capillary electrophoresis and off-line capillary electrophoresis/electrospray ionization-quadrupole time of flight-tandem mass spectrometry approach for ganglioside analysis

Alina D. Zamfir; Željka Vukelić; Jasna Peter-Katalinić

A systematic study for the optimization and implementation of high‐performance capillary electrophoresis (HPCE) in conjunction with negative ion electrospray ionization‐quadrupole time of flight‐tandem mass spectrometry (ESI‐QTOF‐MS/MS) for the analysis of complex glycolipids is described. The performance of the capillary electrophoresis (CE) and off‐line CE/ESI‐QTOF‐MS approach has been explored for screening a complex ganglioside mixture from bovine brain. All instrumental and solution parameters demonstrated to require special adjustment and to have the most substantial effect on the CE separation, abundance of product ions produced in a low‐energy collision‐induced dissociation (CID) process and their detection by MS/MS, when attempting to identify and sequence single ganglioside molecular species from CE eluted fractions. Upon optimization of the experimental parameters, an efficient methodology emerged providing the general basic requirements for combined CE/ESI‐MS analysis of this type of complex glycoconjugate.


Journal of Chromatography A | 2000

Capillary electrophoresis and off-line capillary electrophoresis-electrospray ionization quadrupole time-of-flight tandem mass spectrometry of carbohydrates

Alina D. Zamfir; Simone König; Jörg Althoff; Jasna Peter-Katalinc

The use of off-line high-performance capillary electrophoresis in connection with nanospray electrospray ionization quadrupole time-of-flight tandem mass spectrometry for identification of complex carbohydrates of biological origin is presented. The method was applied to the identification of O-glycosylated amino acids and -glycopeptides from the urine of patients suffering from a hereditary disease - N-acetylhexosaminidase deficiency. Structural elements typical for O-glycosylation of proteins, like expression of core 1 and 2 type O-glycans with different numbers of N-acetyllactosaminyl repeats and different degrees of sialylation, can be directly detected.


European Journal of Mass Spectrometry | 2009

High-throughput analysis of gangliosides in defined regions of fetal brain by fully automated chip-based nanoelectrospray ionization multi-stage mass spectrometry

Alina Serb; Catalin Schiopu; Corina Flangea; Željka Vukelić; Eugen Sisu; Leon Zagrean; Alina D. Zamfir

Gangliosides (GGs), a large group of sialylated glycosphingolipids, are considered biomarkers of human brain development, aging and certain diseases. Determination of individual GG components in complex mixtures extracted from a human brain represents a fundamental prerequisite for correlating their specificity with the specialized function of each brain area. In the context of modern glycomics, detailed investigation of GG expression and structure in human brain requires a continuous development and application of innovative methods able to improve the quality of data and speed of analysis. In this work, for the first time, a high-throughput mapping and sequencing of gangliosides in human fetal brain was performed by a novel mass spectrometry (MS)-based approach developed recently in our laboratory. Three GG mixtures extracted and purified from different regions of the same fetal brain in the 36th gestational week: frontal neocortex (NEO36), white matter of the frontal lobe (FL36) and white matter of the occipital lobe (OL36) were subjected to comparative high-throughput screening and multi-stage fragmentation by fully automated chip-based nanoelectrospray ionization (nanoESI) high capacity ion trap (HCT) MS. Using this method, in only a few minutes of signal acquisitions, over 100 GG and asialo-GG species were detected and identified in the three mixtures. Obtained data revealed for the first time that differences in GG expression in human fetal brain are dependent on phylogenetic development rather than topographic factors. While a significant variation of GG distribution in NEO36 vs FL36 was observed, no significant differences in GG expression in white matter of frontal vs occipital lobe were detected. Additionally, the largest number of species was identified in NEO36, which correlates with the functional complexity of neocortex as the newest brain region. In the last stage of analysis, using MS2–MS3 molecular ion fragmentation at variable amplitudes, a NEO36-associated GD1b isomer could clearly be discriminated. Present results indicate that the combination of fully automated chipESI with HCT MS n is able to provide ultra-fast, sensitive and reliable analyses of complex lipid-linked carbohydrates from which the pattern of their expression and structure in a certain type of bio-matrix can be determined.


Analytical Chemistry | 2016

Electrospray Ionization Ion Mobility Mass Spectrometry of Human Brain Gangliosides

Mirela Sarbu; Adrian C. Robu; Roxana M. Ghiulai; Željka Vukelić; David E. Clemmer; Alina D. Zamfir

The progress of ion mobility spectrometry (IMS), together with its association to mass spectrometry (MS), opened new directions for the identification of various metabolites in complex biological matrices. However, glycolipidomics of the human brain by IMS MS represents an area untouched up to now, because of the difficulties encountered in brain sampling, analyte extraction, and IMS MS method optimization. In this study, IMS MS was introduced in human brain ganglioside (GG) research. The efficiency of the method in clinical glycolipidomics was demonstrated on a highly complex mixture extracted from a normal fetal frontal lobe (FL37). Using this approach, a remarkably rich molecular ion pattern was discovered, which proved the presence of a large number of glycoforms and an unpredicted diversity of the ceramide chains. Moreover, the results showed for the first time the occurrence of GGs in the human brain with a much higher degree of sialylation than previously reported. Using IMS MS, the entire series starting from mono- up to octasialylated GGs was detected in FL37. These findings substantiate early clinical reports on the direct correlation between GG sialylation degree and brain developmental stage. Using IMS CID MS/MS, applied here for the first time to gangliosides, a novel, tetrasialylated O-GalNAc modified species with a potential biomarker role in brain development was structurally characterized. Under variable collision energy, a high number of sequence ions was generated for the investigated GalNAc-GQ1(d18:1/18:0) species. Several fragment ions documented the presence of the tetrasialo element attached to the inner Gal, indicating that GalNAc-GQ1(d18:1/18:0) belongs to the d series.


Glycoconjugate Journal | 2014

Early stage fetal neocortex exhibits a complex ganglioside profile as revealed by high resolution tandem mass spectrometry

Roxana M. Ghiulai; Mirela Sarbu; Željka Vukelić; Constantin Ilie; Alina D. Zamfir

In this study we report on the first mass spectrometric (MS) investigation of gangliosides and preliminary assessment of the expression and structure in normal fetal neocortex in early developmental stages: 14th (Neo14) and 16th (Neo16) gestational weeks. Ganglioside analysis was carried out using a hybrid quadrupole time-of-flight (QTOF) MS with direct sample infusion by nanoelectrospray ionization (nanoESI) in the negative ion mode. Under optimized conditions a large number of glycoforms i.e. 75 in Neo14 and 71 in Neo16 mixtures were identified. The ganglioside species were found characterized by a high diversity of the ceramide constitution, an elevated sialylation degree (up to pentasialylated gangliosides-GP1) and sugar cores modified by fucosylation (Fuc) and acetylation (O-Ac). Direct comparison between Neo14 and Neo16 revealed a prominent expression of monosialylated structures in the Neo16 as well as the presence of a larger number of polysialylated species in Neo14 which constitutes a clear marker of rapid development-dependant changes in the sialylation. Also the MS screening results highlighted that presumably O-acetylation process occurs faster than fucosylation. CID MS/MS under variable collision energy applied for the first time for structural analysis of a fucosylated pentasialylated species induced an efficient fragmentation with generation of ions supporting Fuc-GP1d isomer in early stage fetal brain neocortex.


Central European Journal of Chemistry | 2010

Qualitative and quantitative analysis of gallic acid in Alchemilla vulgaris, Allium ursinum, Acorus calamus and Solidago virga-aurea by chip-electrospray ionization mass spectrometry and high performance liquid chromatography.

Dumitru Condrat; Cristina Mosoarca; Alina D. Zamfir; Florin Crişan; Maria R. Szabo; Alfa Xenia Lupea

AbstractThis study presents the results obtained from qualitative and quantitative analysis of gallic acid from hydro-alcoholic extracts (methanol, ethanol) of plants from Plantae regnum. Plant qualitative analysis was performed using a novel mass spectrometric (MS) method based on fully automated chip-nanoelectrospray ionization (nanoESI) high capacity ion trap (HCT) while quantitative analysis was carried out by high performance liquid chromatography (HPLC). These methods were applied to Alchemilla vulgaris — common lady’s-mantle (aerial part), Allium ursinum — bear’s garlic (leaves), Acorus calamus — common sweet flag (roots), Solidago virga-aurea — goldenrod (aerial part). Obtained results indicated that methanol extracts (96%, 80%) have a gallic acid content ranging between 0.0011–0.0576 mg mL−1 extract while the ethanol extracts (96%, 60%) exhibit a gallic acid concentration that varies between 0.0010–0.0182 mg mL−1 extract.n


Carbohydrate Research | 2014

Automated chip-nanoelectrospray mass spectrometry for glycourinomics in Schindler disease type I.

Mirela Sarbu; Adrian C. Robu; Jasna Peter-Katalinić; Alina D. Zamfir

In this study an integrative mass spectrometry (MS) approach based on fully automated chip-nanoelectrospray quadrupole time-of-flight was optimized and applied for the discovery and structural characterization of O-glycopeptides in a fraction from the urine of a patient diagnosed with Schindler disease type I. A mixture of O-glycopeptides extracted and purified from an age matched healthy subject served as the control. 49 glycoforms were discovered in the investigated urine fraction from Schindler disease versus only 14 in control urine. Structures with relevant biological significance, previously not described, such as O-fucosylated tetrasaccharides and chains up to pentadecamers O-linked to serine, threonine, or threonine-proline were identified in the pathological urine and characterized by tandem MS (MS/MS). A number of 29 species discovered here, most of which with long chain glycans, were not previously reported as associated to this condition. All glycopeptides were detected in only 1 min analysis time, with a sample consumption situated in the femtomole range.


Analytical Biochemistry | 2017

Assessment of ganglioside age-related and topographic specificity in human brain by Orbitrap mass spectrometry

Mirela Sarbu; Liana Dehelean; Cristian V.A. Munteanu; Željka Vukelić; Alina D. Zamfir

The gangliosides (GGs) of the central nervous system (CNS) exhibit age and topographic specificity and these patterns may correlate with the functions and pathologies of the brain regions. Here, chloroform extraction, nanoelectrospray (nanoESI) negative ionization, together with Orbitrap high resolution mass spectrometry (MS) determined the topographic and age-related GG specificity in normal adult human brain. Mapping of GG mixtures extracted from 20 to 82 year old frontal and occipital lobes revealed besides a decrease in the GG number with age, a variability of sialylation degree within the brain regions. From the 111 species identified, 105 were distinguished in the FL20, 74 in OL20, 46 in FL82 and 56 in OL82. The results emphasize that within the juvenile brain, GG species exhibit a higher expression in the FL than in OL, while in the aged brain the number of GG species is higher in the OL. By applying MS/MS analysis, the generated fragment ions confirmed the incidence of GT1c (d18:1/18:0) and GT1c (d18:1/20:0) in the investigated samples. The present findings are of major value for further clinical studies carried out using Orbitrap MS in order to correlate gangliosides with CNS disorders.

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Mirela Sarbu

Aurel Vlaicu University of Arad

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Hans Kresse

University of Münster

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Adrian C. Robu

Aurel Vlaicu University of Arad

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