Papers by cosima damiana calvano
International journal of molecular sciences, May 29, 2024
While edible algae might seem low in fat, the lipids they contain are crucial for good health and... more While edible algae might seem low in fat, the lipids they contain are crucial for good health and preventing chronic diseases. This study introduces a binary matrix to analyze all the polar lipids in both macroalgae (Wakame-Undaria pinnatifida, Dulse-Palmaria palmata, and Nori-Porphyra spp.) and microalgae (Spirulina-Arthrospira platensis, and Chlorella-Chlorella vulgaris) using matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The key lies in a new dual matrix made by combining equimolar amounts of 1,5-diaminonaphthalene (DAN) and 9-aminoacridine (9AA). This combination solves the limitations of single matrices: 9AA is suitable for sulfur-containing lipids and acidic phospholipids, while DAN excels as an electron-transfer secondary reaction matrix for intact chlorophylls and their derivatives. By employing the equimolar binary matrix, a wider range of algal lipids, including free fatty acids, phospholipids, glycolipids, pigments, and even rare arsenosugarphospholipids were successfully detected, overcoming drawbacks related to ion suppression from readily ionizable lipids. The resulting mass spectra exhibited a good signalto-noise ratio at a lower laser fluence and minimized background noise. This improvement stems from the binary matrix's ability to mitigate in-source decay effects, a phenomenon often encountered for certain matrices. Consequently, the data obtained are more reliable, facilitating a faster and more comprehensive exploration of algal lipidomes using high-throughput MALDI-MS/MS analysis.
Molecules
Reversed-phase liquid chromatography and electrospray ionization with Fourier-transform single an... more Reversed-phase liquid chromatography and electrospray ionization with Fourier-transform single and tandem mass spectrometry (RPLC-ESI-FTMS and FTMS/MS) were employed for the structural characterization of oleocanthal (OLEO) and oleacin (OLEA), two of the most important bioactive secoiridoids occurring in extra virgin olive oils (EVOOs). The existence of several isoforms of OLEO and OLEA was inferred from the chromatographic separation, accompanied, in the case of OLEA, by minor peaks due to oxidized OLEO recognized as oleocanthalic acid isoforms. The detailed analysis of the product ion tandem MS spectra of deprotonated molecules ([M-H]−) was unable to clarify the correlation between chromatographic peaks and specific OLEO/OLEA isoforms, including two types of predominant dialdehydic compounds, named Open Forms II, containing a double bond between carbon atoms C8 and C10, and a group of diasteroisomeric closed-structure (i.e., cyclic) isoforms, named Closed Forms I. This issue was a...
Organohalogen compounds, 2015
Calaprice C, Calvano CD, Zambonin C, Focant JF* University of Liège, Chemistry Institute (B6c), M... more Calaprice C, Calvano CD, Zambonin C, Focant JF* University of Liège, Chemistry Institute (B6c), Mass Spectrometry Laboratory, Organic & Biological Analytical Chemistry Group, Allée du Six Août, 11, Quartier Agora B-4000 Sart-Tilman, (Liège) – Belgium; Politecnico di Bari, Dipartimento di Ingegneria Civile, Ambientale, del Territorio, Edile e di Chimica, Via Orabona, 4, 70125 Bari – Italy; Università degli studi di Bari, Dipartimento di Chimica, via Orabona 4, 70125 Bari – Italy *[email protected]
In the past decade, hydrophilic interaction liquid chromatography (HILIC) has emerged as an effic... more In the past decade, hydrophilic interaction liquid chromatography (HILIC) has emerged as an efficient alternative to reversed-phase chromatography (RPC) for the analysis of phospholipid (PL) mixtures based on mass spectrometric detection. Since the separation of PL by HILIC is chiefly based on their headgroup, the mass spectrum of each class can be obtained by spectral averaging under the corresponding HILIC band. Using experimental m/z values resulting from high mass resolution/accuracy instruments, the sum compositions of PL in a specific class can be thus inferred but partial overlapping may occur between signals related to the M + 0 isotopologue of one species and the M + 2/M + 4 isotopologues of species having one/two more C═C bonds in their chemical structures. Here, an automated workflow, named LIPIC (lipid isotopic pattern interference correction), is proposed to account for such interferences. Starting from the experimentally verified assumption that peaks in isotope patterns are Gaussian, LIPIC predicts, as a function of m/z ratio, signal intensities due to M + 2 and M + 4 isotopologues of species with one or two more C = C bonds than the target one and calculates the corrected intensity for the M + 0 isotopologue of the latter. Thanks to an iterative procedure, the suggested algorithm compensates also for slight shifts occurring between experimental and theoretical m/z ratios related to isotopologue peaks. Examples of applications to simulated and experimental mass spectra of two PL classes, i.e., phosphatidylcholines (PC) and cardiolipins (CL), emphasize the increased extent of correction at the increase of molecular masses of involved species.
Methods in Molecular Biology, 2020
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is largely recognized ... more Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is largely recognized as an important tool in the analysis of many biomolecules such as proteins and peptides. The MS analysis of digested peptides to identify a protein or some of its modifications is a key step in proteomics. MALDI-MS is well suited for the peptide mass fingerprinting (PMF) technique, as well as selected fragmentation of various precursors using collisional-induced dissociation (CID) or post-source decay (PSD).In the last few years, MALDI-MS has played a significant role in food chemistry, especially in the detection of food adulterations, characterization of food allergens, and investigation of protein structural modifications induced by various industrial processes that could be an issue in terms of food quality and safety.Here, we present simple extraction protocols of allergenic proteins in food commodities such as milk, egg, hazelnut , and lupin seeds. Classic bottom-up approaches based on Sodium Dodecyl Sulphate (SDS) gel electrophoresis separation followed by in-gel digestion or direct in-solution digestion of whole samples are described. MALDI-MS and MS /MS analyses are discussed along with a comparison of data obtained by using the most widespread matrices for proteomic studies, namely, α-cyano-4-hydroxy-cinnamic acid (CHCA) and α-cyano-4-chloro-cinnamic acid (CClCA). The choice of the most suitable MALDI matrix is fundamental for high-throughput screening of putative food allergens.
Mass Spectrometry Reviews, 2021
Surfactants are surface‐active agents widely used in numerous applications in our daily lives as ... more Surfactants are surface‐active agents widely used in numerous applications in our daily lives as personal care products, domestic, and industrial detergents. To determine complex mixtures of surfactants and their degradation products, unselective and rather insensitive methods, based on colorimetric and complexometric analyses are no longer employable. Analytical methodologies able to determine low concentration levels of surfactants and closely related compounds in complex matrices are required. The recent introduction of robust, sensitive, and selective mass spectrometry (MS) techniques has led to the rapid expansion of the surfactant research field including complex mixtures of isomers, oligomers, and homologues of surfactants as well as their chemically and biodegradation products at trace levels. In this review, emphasis is given to the state‐of‐the‐art MS‐based analysis of surfactants and their degradation products with an overview of the current research landscape from tradit...
The chemical identity of arsenosugar phospholipids (As-PL) as mono-(i.e., lyso, L-As-PL) and diac... more The chemical identity of arsenosugar phospholipids (As-PL) as mono-(i.e., lyso, L-As-PL) and diacyl-arsenosugar PL in four edible and common marine alga samples, such as nori (Porphyra spp.), wakame (Undaria pinnatifida), dulse (Palmaria palmata), and kombu (Saccharina japonica), was successfully investigated. Adopting negative polarity electrospray ionization (ESI), not common for As-PL, conjugated with hydrophilic interaction liquid chromatography (HILIC) and mass spectrometry (MS), performed either at low resolution using a linear ion trap (LIT) with sequential MS n (n = 2, 3) or at high resolution using a high-resolution/high-accuracy Fourier-transform MS (FTMS), based on an orbital trap instrument, more than 20 As-PL and 2 L-As-PL species were identified. The absence of As-PL standard compounds encouraged us to generate an in-house-built database of As-PL/L-As-PL for a rapid and simple classification. Despite their compositional diversity, tandem MS of deprotonated As-PL and L-As-PL ([M -H] -) demonstrated the occurrence of a highly diagnostic product ion at m/z 389.0 ([AsC 10 H 19 O 9 P] -). The fatty acid composition and distribution of As-PL were easily assigned on the basis of the ratio intensity between sn-1 and sn-2 product ions. Indeed, the preferential formation of [R 1 C 3 H 5 O 4 P] -ions over [R 2 C 3 H 5 O 4 P] -ions, both containing the glycerol backbone, enabled the regiochemical assignment of As-PL. These outcomes were confirmed by MS n (n = 2, 3) analyses and using sn-1-and sn-2regioselective hydrolase enzymes (i.e., phospholipases A 1 and A 2 ). The predominant As-PL's in samples of nori (red alga), wakame, and kombu (both brown algae) were identified as containing palmitic acyl chains (i.e., As-PL958 (As-PL 16:0/16:0) with ca. 66 ± 3, 82 ± 4, and 58 ± 3% as relative abundances, respectively), while the main species in dulse (red alga) samples was As-PL982 (As-PL 18:1/16:1) at ca. 38 ± 3%.
Journal of Agricultural and Food Chemistry, 2020
Reversed-phase liquid chromatography with electrospray ionization-high resolution/accuracy Fourie... more Reversed-phase liquid chromatography with electrospray ionization-high resolution/accuracy Fourier-transform mass spectrometry (RPC-ESI-FTMS) and chemometrics were exploited to evaluate the influence of horizontal centrifugation by two-or three-phase decanters on the content of major phenolic secoiridoids in extra-virgin olive oils (EVOOs). Despite the occurrence of other potential sources of variability typical of commercial olive oils, horizontal centrifugation was found to play a primary role, with a general increase of secoiridoids content occurring when twophase decanters were used. As emphasized by Principal Component Analysis (PCA), the increase involved preferentially oleacin and oleocanthal, when oxidative deterioration was purposely minimized during and/or after production, and oleuropein and ligstroside aglycones, when no vertical centrifugation was performed at the end of the productive cycle. The influence of the type of horizontal centrifugation was emphasized also by the elaboration of RPC-ESI-FTMS data based on Hierarchical Cluster Analysis (HCA) and Linear Discriminating Analysis (LDA).
Rapid Communications in Mass Spectrometry, 2019
RATIONALE: Several bioactive compounds, including phenolic acids and secoiridoids, are transferre... more RATIONALE: Several bioactive compounds, including phenolic acids and secoiridoids, are transferred from olive drupes to olive oil during the first stage of production. Here, the characterization of these low molecular weight (LMW) compounds in olive oil and in closely related processing materials, like olive leaves (OL) and olive mill wastewaters (OMW), was faced up, for the first time, by matrix-assisted laser desorption ionization (MALDI) time-offlight mass spectrometry (TOF MS). A novel binary matrix composed of 1,8-bis(tetramethylguanidino)naphthalene (TMGN) and 9-aminoacridine (9AA), (1:1 molar ratio), displaying excellent ionization properties at low levels of laser energy, was employed in reflectron negative ion mode by a MALDI TOF/TOF system equipped with a neodymium-doped yttrium lithium fluoride (Nd:YLF) laser (345 nm). MS/MS experiments were performed by using ambient air as the collision gas. Four major secoiridoids typically present in olive oil, i.e., the aglycones of oleuropein and ligstroside, and oleacein and olecanthal at m/z 377.1, 361.1, 319.1 and 303.1, respectively, were detected in virgin olive oil (VOO) extracts, along with some of their chemical/enzymatic hydrolysis by-products, such as elenolic (m/z 241.1), decarboxymethylelenolic acids (m/z 183.1) and hydroxytyrosol (m/z 153.1). Besides oleuropein aglycone and oleacein, hydroxylated derivatives of decarboxymethyl-elenolic acid and hydroxytyrosol were evidenced in OMW. While oleuropein was confirmed in OL extracts, several interesting phenolic compounds, including hydroxytyrosol, were recognized in OMW. The proposed approach based on the use of a novel binary matrix for MALDI MS/MS analyses of LMW bioactive compounds can be considered a promising analytical tool for a rapid screening of the phenolic fraction in olive oils and related processing wastes.
Analytical and Bioanalytical Chemistry, 2018
Since its introduction in 1980s, matrix-assisted laser desorption/ionization mass spectrometry (M... more Since its introduction in 1980s, matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has gained a prominent role in the analysis of high molecular weight biomolecules such as proteins, peptides, oligonucleotides and polysaccharides. Its application to low molecular weight compounds has remained for long time challenging due to the spectral interferences produced by conventional organic matrices in the low m/z window. To overcome this problem, specific sample preparation such as analyte/matrix derivatization, addition of dopants or sophisticated deposition technique especially useful for imaging experiments, have been proposed. Alternative approaches based on second generation (rationally designed) organic matrices, ionic liquids and inorganic matrices including metallic nanoparticles, have been the object of intense and continuous research efforts. Definite evidences are now provided that MALDI MS represents a powerful and invaluable analytical tool also for small molecules, including their quantification, thus opening new exciting applications in metabolomics and imaging mass spectrometry. This review is intended to offer a concise critical overview of the most recent achievements about MALDI matrices capable of specifically address the challenging issue of small molecules analysis.
Analytical and bioanalytical chemistry, 2016
The most recent advances in the identification and determination of organic constituents in paint... more The most recent advances in the identification and determination of organic constituents in paintings and other polychrome objects using mass spectrometry (MS)-based techniques are reviewed. The latest achievements in gas chromatography (GC)-MS and pyrolysis (Py-) GC-MS are mainly related to sample pretreatment protocols and to the employment of double-shot or laser desorption pyrolysis, respectively. MS techniques based on soft ionization methods such as matrix assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) are discussed. So far, MALDI and ESI MS have been mainly used in the characterization of proteinaceous materials, but further applications are definitely emerging, e.g., in the fields of lipids, resins, and organic colorants analysis. Chemical imaging by time-of-flight secondary ion mass spectrometry (TOF SIMS), formerly applied to the detection and localization of lipid binders and inorganic materials, has been recently extended to proteins. Fina...
Food Authentication, 2017
Journal of Mass Spectrometry, 2016
4‐Chloro‐α‐cyanocinnamic acid (ClCCA) is a very useful matrix able to give the protonated adduct ... more 4‐Chloro‐α‐cyanocinnamic acid (ClCCA) is a very useful matrix able to give the protonated adduct [M+H]+ of intact cyanocobalamin (CNCbl) as the base peak (m/z 1355.58) in matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry (MS). The only fragment observed is [M‐CN + H]+• formed through the facile •CN neutral loss reflecting the fairly low Co–C bond energy. All other investigated proton transfer matrices, including α‐cyano‐4‐hydroxycinnamic acid, para‐nitroaniline and 2,5‐dihydroxybenzoic acid, give rise to a complete decyanation of CNCbl with concomitant formation of [M‐CN + H]+•, [M‐CN + Na]+• and [M‐CN + K]+• adducts at m/z 1329.57, 1351.55 and 1367.51, respectively. Depending on the matrix used, a variable degree of fragmentation involving the α‐side axial ligand was observed. A plausible explanation of the specific behaviour of 4‐chloro‐α‐cyanocinnamic acid as a soft matrix is discussed. Tandem mass spectra of both [M + H]+ and [M‐CN + H]+• ions were obtained a...
Journal of Mass Spectrometry, 2016
In this work, novel hybrid nanostructured surfaces, consisting of dense arrays of silicon nanowir... more In this work, novel hybrid nanostructured surfaces, consisting of dense arrays of silicon nanowires (SiNWs) functionalized by Ag nanoparticles (AgNP/SiNWs), were used for the laser desorption/ionization time‐of‐flight mass spectrometry (LDI‐TOF MS) analysis of some typical unsaturated food components (e.g. squalene, oleic acid) to assess their MS performance. The synthesis of the novel platforms is an easy, cost‐effective process based on the maskless wet‐etching preparation at room temperature of SiNWs followed by their decoration with AgNPs, produced by pulsed laser deposition. No particular surface pretreatment or addition of organic matrixes/ionizers was necessary. Moreover, oil extracts (e.g. extra virgin olive oil, peanut oil) could be investigated on AgNP/SiNWs surfaces, revealing their different MS profiles. It was shown that such substrates operate at reduced laser energy, typically generating intense silver cluster ions and analyte adducts. A comparison with bare SiNWs was...
Proceedings of the National Academy of Sciences of the United States of America, Jan 22, 2015
Prior studies show that oxytocin (Oxt) and vasopressin (Avp) have opposing actions on the skeleto... more Prior studies show that oxytocin (Oxt) and vasopressin (Avp) have opposing actions on the skeleton exerted through high-affinity G protein-coupled receptors. We explored whether Avp and Oxtr can share their receptors in the regulation of bone formation by osteoblasts. We show that the Avp receptor 1α (Avpr1α) and the Oxt receptor (Oxtr) have opposing effects on bone mass: Oxtr(-/-) mice have osteopenia, and Avpr1α(-/-) mice display a high bone mass phenotype. More notably, this high bone mass phenotype is reversed by the deletion of Oxtr in Oxtr(-/-):Avpr1α(-/-) double-mutant mice. However, although Oxtr is not indispensable for Avp action in inhibiting osteoblastogenesis and gene expression, Avp-stimulated gene expression is inhibited when the Oxtr is deleted in Avpr1α(-/-) cells. In contrast, Oxt does not interact with Avprs in vivo in a model of lactation-induced bone loss in which Oxt levels are high. Immunofluorescence microscopy of isolated nucleoplasts and Western blotting an...
Chemosphere, 2015
This is the first report on levels of Dechloranes in European Food and feed. A specific GC-QQQMS/... more This is the first report on levels of Dechloranes in European Food and feed. A specific GC-QQQMS/MS method was developed. Most selected Dechloranes were detected in European food and feed. Dechloranes are at the pg/g fat levels in food. A Dechlorane dietary intake was estimated for the Belgian population.
Analytical and Bioanalytical Chemistry, 2015
Chlorophylls (Chls) are important pigments responsible for the characteristic green color of chlo... more Chlorophylls (Chls) are important pigments responsible for the characteristic green color of chloroplasts in algae and plants. In this study, 1,5-diaminonaphthalene (DAN) was introduced as an electron transfer secondary reaction matrix for the identification of intact chlorophylls and their derivatives, by matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). DAN was proved to drastically outperform conventional matrices such as αcyano-4-hydroxycinnnamic acid, dithranol, antracene, and even terthiophene, since loss of the metal ion and fragmentation of the phytol-ester linkage are negligible. Absence of significant fragmentation of radical cations of Chls a and b at m/z 892.529 and 906.513, respectively, makes MALDI MS capable of following natural degradation of intact porphyrinbased pigments whose initial steps are just represented by demetalation and dephytylation. Chl by-products, such as pyropheophytins, have been identified in dried tea leaves showing the potential of MALDI MS to follow chlorophyll biotransformation occurring in processed foodstuffs. Finally, preliminary results show the potential of MALDI MS to detect illegal vegetable oil re-greening practices.
Journal of agricultural and food chemistry, 2014
Adulteration of extra-virgin olive oil (EVOO) with hazelnut oil (HO) is an illegal practice that ... more Adulteration of extra-virgin olive oil (EVOO) with hazelnut oil (HO) is an illegal practice that could have severe health consequences for consumers due to the possible exposure to hidden hazelnut allergens. Here, matrix-assisted laser-desorption/ionization (MALDI) mass spectrometry (MS) was used as a rapid and sensitive technique for the detection of a low concentration of hazelnut proteins in oil samples. Different protocols were tested for protein extraction, and the most efficient (cold acetone) was applied to HO and EVOO adulterated with HO. The subsequent in-solution tryptic digestion of protein extracts and MALDI-MS analysis, using α-cyano-4-chlorocinnamic acid as matrix, allowed the detection of stable hazelnut peptide markers (i.e., the m/z ions 1002.52, 1356.71, 1394.70, 1440.81, 1453.85, 1555.76, 1629.83, 1363.73, and 1528.67) attributable to the main hazelnut proteins Cor a 9, Cor a 11, and Cor a 1. Thus, the approach might allow the direct detection of specific hazelnut...
Talanta, 2015
The simultaneous identification of lipids and proteins by matrix assisted laser desorption ioniza... more The simultaneous identification of lipids and proteins by matrix assisted laser desorption ionizationmass spectrometry (MALDI-MS) after direct on-plate processing of micro-samples supported on colloidal graphite is demonstrated. Taking advantages of large surface area and thermal conductivity, graphite provided an ideal substrate for on-plate proteolysis and lipid extraction. Indeed proteins could be efficiently digested on-plate within 15 min, providing sequence coverages comparable to those obtained by conventional in-solution overnight digestion. Interestingly, detection of hydrophilic phosphorylated peptides could be easily achieved without any further enrichment step. Furthermore, lipids could be simultaneously extracted/identified without any additional treatment/processing step as demonstrated for model complex samples such as milk and egg. The present approach is simple, efficient, of large applicability and offers great promise for protein and lipid identification in very small samples.
Proceedings of the National Academy of Sciences, 2014
Significance We have shown previously that oxytocin (Oxt), other than regulating lactation and so... more Significance We have shown previously that oxytocin (Oxt), other than regulating lactation and social bonding, is a potent stimulator of bone formation by the osteoblast. Here, we present evidence that this action is exerted through the nuclear localization of the Oxt receptor (Oxtr). Our findings prompt additional studies into the contribution of nuclear Oxtr signaling in regulating lactation and social bonding.
Uploads
Papers by cosima damiana calvano