Roberto Samperi - Academia.edu (original) (raw)

Papers by Roberto Samperi

Research paper thumbnail of Development and validation of a liquid chromatography/atmospheric pressure photoionization-tandem mass spectrometric method for the analysis of mycotoxins subjected to commission regulation (EC) No. 1881/2006 In cereals

Journal of Chromatography A, 2010

A sensitive and reliable liquid chromatography/photoionization (APPI) tandem mass spectrometry me... more A sensitive and reliable liquid chromatography/photoionization (APPI) tandem mass spectrometry method has been developed for determining nine selected mycotoxins in wheat and maize samples. The analytes were chosen on the basis of the mycotoxins under EU Commission Regulation (EC) No. 1881/2006, i.e., deoxynivalenol (DON), zearalenone (ZON), aflatoxins (AFs), and ochratoxin A (OTA), and considering the possibility of a near future

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Research paper thumbnail of Liquid chromatography/tandem mass spectrometric confirmatory method for determining aflatoxin M1 in cow milk

Journal of Chromatography A, 2006

A liquid chromatography/electrospray (ESI)–tandem mass spectrometric method for the measurement o... more A liquid chromatography/electrospray (ESI)–tandem mass spectrometric method for the measurement of aflatoxin M1 (AFM1) in milk is described. Milk sample after protein precipitation with acetone was cleaned-up with a Carbograph-4 cartridge. Performances of the ESI source were compared with those of the atmospheric pressure photoionization source (APPI). Although a method quantification limit (MQL) of 6ng/kg could be achieved operating with

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Research paper thumbnail of Reversed-phase high-performance liquid chromatographic determination of linear alkylbenzene sulphonates, nonylphenol polyethoxylates and their carboxylic biotransformation products

Journal of Chromatography A, 1993

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Research paper thumbnail of Ultra-high-performance liquid chromatography-tandem mass spectrometry for the analysis of free and conjugated natural estrogens in cow milk without deconjugation

Analytical and Bioanalytical Chemistry

A sensitive liquid chromatography/electrospray ionization-tandem mass spectrometry method for the... more A sensitive liquid chromatography/electrospray ionization-tandem mass spectrometry method for the determination of free and conjugated estrogens in cow milk is described. Milk samples were diluted with water and then extracted and cleaned up by solid-phase extraction using graphitized carbon black as adsorbent material, without any enzymatic cleavage, derivatization, and/or protein precipitation step. Two fractions were collected (free and conjugated estrogens) and analyzed separately. Mass spectrometry analysis was performed in negative ion mode using selected reaction monitoring acquisition mode. For all compounds, the coefficients of determination (R (2)) ranged between 0.9892 and 0.9997. Analytical recoveries were in the range of 86-109 % for free estrogens and 85-118 % for conjugates, with relative standard deviations below 10 %, and the method detection limit ranged between 3 and 80 ng L(-1). Finally, the developed method was used to determine the presence of free and conjugat...

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Research paper thumbnail of Liquid chromatography/tandem mass spectrometry determination of organophosphorus flame retardants and plasticizers in drinking and surface waters

Rapid Communications in Mass Spectrometry

A liquid chromatography/electrospray ionization tandem mass spectrometric method for analyzing or... more A liquid chromatography/electrospray ionization tandem mass spectrometric method for analyzing organophophorus flame retardants and plasticizers in drinking and environmental waters was developed. Five alkyl phosphates, three chlorinated alkyl phosphates, two aryl phosphate and triphenylphosphine oxide were selected for this study. These compounds were extracted from water samples by a hydrophilic polymeric solid-phase extraction cartridge. Accuracy and precision were evaluated analyzing 0.5 L of water samples spiked at concentrations of 10 and 100 ng/L for drinking water and at 300 and 1000 ng/L for river water. Except for trimethyl phosphate, analyte recoveries were better than 80%, and were not dependent on the type of aqueous matrix in which they were dissolved. At the spike levels considered, within-day precision was between 3 and 12% for tap water and between 4 and 14% for river water, and estimated method quantification limits ranged from 0.2 to 3.9 ng/L. A short survey condu...

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Research paper thumbnail of Peptidome characterization and bioactivity analysis of donkey milk

Journal of Proteomics, 2015

Donkey milk is an interesting commercial product for its nutritional values, which make it the mo... more Donkey milk is an interesting commercial product for its nutritional values, which make it the most suitable mammalian milk for human consumption, and for the bioactivity associated with it and derivative products. To further mine the characterization of donkey milk, an extensive peptidomic study was performed. Two peptide purification strategies were compared to remove native proteins and lipids and enrich the peptide fraction. In one case the whole protein content was precipitated by organic solvent using cold acetone. In the other one the precipitation of the most abundant milk proteins, caseins, was performed under acidic conditions by acetic acid at pH4.6, instead. The procedures were compared and proved to be partially complementary. Considered together they provided 1330 peptide identifications for donkey milk, mainly coming from the most abundant proteins in milk. The bioactivity of the isolated peptides was also investigated, both by angiotensin-converting-enzyme inhibitory and antioxidant activity assays and by bioinformatics, proving that the isolated peptides did have the tested biological activities. The rationale behind this study is that peptides in food matrices often play an important biological role and, despite the extensive study of the protein composition of different samples, they remain poorly characterized. In fact, in a typical shotgun proteomics study endogenous peptides are not properly characterized. In proteomics workflows one limiting point is the isolation process: if it is specific for the purification of proteins, it often comprises a precipitation step which aims at isolating pure protein pellets and remove unwonted interferent compounds. In this way endogenous peptides, which are not effectively precipitated as well as proteins, are removed too and not analyzed at the end of the process. Moreover, endogenous peptides do often originate from precursor proteins, but in phenomena which are independent of the shotgun digestion protocol, thus they can be obtained from cleavage specificities other than trypsin's, which is the main proteolytic enzyme employed in proteomic experiments. For this reason, in the end, database search will not be effective for identification of these peptides, thus the need to provide different workflows for peptide analysis. In the work presented in this paper this issue is considered for the first time for the analysis of the peptides isolated in donkey milk samples, which have been chosen for its nutritional interest. This study provides additional knowledge on this milk, already characterized by traditional proteomics studies and peptidomic studies after simulated digestion. This type of study is not just a description of the naturally occurring peptidome of a sample, but also represents a starting point to discover and characterize those naturally occurring peptides responsible for the observed bioactivities of biological samples, as in the case of donkey milk, which would remain uncharacterized by other approaches. In this paper an analytical protocol was described for the efficient isolation and purification of peptides in donkey milk, assessing the effect of the purification protocol on the final identifications. Purified peptide samples were also checked to empirically elucidate any ACE inhibitory or antioxidant activity. Finally, the peptidomic results were also further mined by a bioinformatic-driven approach for bioactive peptide identification in the donkey milk samples. In our opinion, the main strengths of this study are related to the improved analytical workflow (either as purification protocol comparison or analytical platform development) which provides a high number of identified peptides, for which the biological significance as potential bioactive peptides has also been investigated.

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Research paper thumbnail of Application of an innovative matrix solid-phase dispersion–solid-phase extraction–liquid chromatography–tandem mass spectrometry analytical methodology to the study of the metabolism of the estrogenic mycotoxin zearalenone in rainbow trout liver and muscular tissue

International Journal of Environmental Analytical Chemistry, 2004

The metabolic fate of the estrogenic mycotoxin zearalenone in rainbow trout is presently unknown.... more The metabolic fate of the estrogenic mycotoxin zearalenone in rainbow trout is presently unknown. In this study, the tissue concentration of zearalenone and its principal metabolites (α-zearalenol and β-zearalenol) was determined. A known amount of zearalenone was administered as a single bolus to ten fish, and the biological tissue concentration was determined at various times following administration. The analytes were

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Research paper thumbnail of Protein Profile of Mature Soybean Seeds and Prepared Soybean Milk

Journal of Agricultural and Food Chemistry, 2014

The soybean (Glycine max (L.) Merrill) is economically the most important bean in the world, prov... more The soybean (Glycine max (L.) Merrill) is economically the most important bean in the world, providing a wide range of vegetable proteins. Soybean milk is a colloidal solution obtained as water extract from swelled and ground soybean seeds. Soybean proteins represent about 35-40% on a dry weight basis and they are receiving increasing attention with respect to their health effects. However, the soybean is a well-recognized allergenic food, and therefore, it is urgent to define its protein components responsible for the allergenicity in order to develop hypoallergenic soybean products for sensitive people. The main aim of this work was the characterization of seed and milk soybean proteome and their comparison in terms of protein content and specific proteins. Using a shotgun proteomics approach, 243 nonredundant proteins were identified in mature soybean seeds.

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Research paper thumbnail of Multiresidue determination of UV filters in water samples by solid-phase extraction and liquid chromatography with tandem mass spectrometry analysis

Journal of Separation Science, 2014

UV filters, contained in sunscreens and other cosmetic products, as well as in some plastics and ... more UV filters, contained in sunscreens and other cosmetic products, as well as in some plastics and industrial products, are nowadays considered contaminants of emerging concern because their widespread and increasing use has lead to their presence in the environment. Furthermore, some UV filters are suspected to have endocrine disruption activity. In the present work, we developed an analytical method based on liquid chromatography with tandem mass spectrometry for the determination of UV filters in tap and lake waters. Sixteen UV filters were extracted from water samples by solid-phase extraction employing graphitized carbon black as adsorbent material. Handling 200 mL of water sample, satisfactory recoveries were obtained for almost all the analytes. The limits of detection and quantification of the method were comparable to those reported in other works, and ranged between 0.7-3.5 and 1.9-11.8 ng/L, respectively; however in our case the number of investigated compounds was larger. The major encountered problem in method development was to identify the background contamination sources and reduce their contribution. UV filters were not detected in tap water samples, whereas the analyses conducted on samples collected from three different lakes showed that the swimming areas are most subject to UV filter contamination.

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Research paper thumbnail of Development of an analytical strategy for the identification of potential bioactive peptides generated by in vitro tryptic digestion of fish muscle proteins

In the last years, food proteins and peptides are attracting great attention because of the emerg... more In the last years, food proteins and peptides are attracting great attention because of the emergence of a new field, that of food-derived bioactive peptides. This paper presents a comparison and evaluation of four different experiments for the identification of sarcoplasmic and myofibrillar fish peptides. This study is aimed at the development of a simple and fast method for the identification of peptides that could arise from fish meat if trypsin was the only digestive enzyme acting on fish meat proteins. In particular, we tested the use of ultrafiltration membranes with a molecular weight cutoff of 3,000 Da. Data analysis has shown that the experiment in which there is neither precipitation nor an ultrafiltration step performed better and allowed the identification of a larger number of peptides and potential antimicrobial peptides (AMPs); this workflow provided 473 and 398 total identified peptides and 44 and 18 AMPs for sarcoplasmic and myofibrillar extracts, respectively. This protocol is found to be faster and more straightforward than the other three tested workflows. The developed strategy could be also useful for other food matrices and could provide information about food quality and safety control.

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Research paper thumbnail of Effect of membrane charge density on the protein corona of cationic liposomes: Interplay between cationic charge and surface area

Applied Physics Letters, 2011

Here we investigate the effect of membrane charge density on the protein corona that forms upon e... more Here we investigate the effect of membrane charge density on the protein corona that forms upon exposure of cationic liposomes to human plasma. We show that the protein corona around cationic liposomes is not uniquely defined by membrane charge density, but it is deeply affected by both the overall cationic charge and the available surface area. Remarkably, changes in protein

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Research paper thumbnail of Comparative analysis of metabolic proteome variation in ascorbate-primed and unprimed wheat seeds during germination under salt stress

Journal of Proteomics, 2014

Seed priming with ascorbic acid improves salt tolerance in durum wheat. For understanding the pot... more Seed priming with ascorbic acid improves salt tolerance in durum wheat. For understanding the potential mechanisms underlying this priming effect a gel-free shotgun proteomic analysis was performed comparing unprimed to ascorbate-primed wheat seed during germination under saline and non-saline conditions. Since seed germination is the result of interplay or cross-talk between embryo and embryo-surrounding tissues, we studied the variation of metabolic proteome in both tissues separately. 167 of 697 identified and 69 of 471 identified proteins increase or decrease in abundance significantly in response to priming and/or salinity compared to untreated, unstressed control in embryo and embryo-surrounding tissues, respectively. In untreated wheat embryo salt stress was accompanied by change in 129 proteins, most of which are belonging to metabolism, energy, disease/defense, protein destination and storage categories. Ascorbate pretreatment prevents and counteracts the effects of salinity upon most of these proteins and changes specifically the abundance of 35 others proteins, most of which are involved in metabolism, protein destination and storage categories. Hierarchical clustering analysis revealed three and two major clusters of protein expression in embryo and embryo-surrounding tissues, respectively. This study opens promising new avenues to understand priming-induced salt tolerance in plants. To clearly understand how ascorbate-priming enhance the salt tolerance of durum wheat during germination, we performed for the first time a comparative shotgun proteomic analysis between unprimed and ascorbate-primed wheat seeds during germination under saline and non-saline conditions. Furthermore, since seed germination is the result of interplay or cross-talk between embryo and embryo-surrounding tissues we analyzed the variation of metabolic proteome in both tissues separately. 1168 proteins exhibiting greater molecular weight diversity (ranging from 5 to 258kDa) were identified. Among them, 167 and 69 proteins were increased or decreased in abundance significantly by priming and/or salinity as compared to control, in embryo and embryo-surrounding tissues respectively. Ascorbate pretreatment alleviates the effects of salinity upon most of these proteins, particularly those involved in metabolism, energy, disease/defense, protein destination and storage functions. Hierarchical clustering analysis revealed three and two major clusters of protein accumulation in embryo and embryo-surrounding tissues, respectively. These results may provide new avenues for understanding and advancing priming-induced salt tolerance in crop plants.

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Research paper thumbnail of List of Contributors

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Research paper thumbnail of Recent trends in matrix solid-phase dispersion

TrAC Trends in Analytical Chemistry, 2013

ABSTRACT The key factors for the success of matrix solid-phase dispersion (MSPD) are its feasibil... more ABSTRACT The key factors for the success of matrix solid-phase dispersion (MSPD) are its feasibility, flexibility, versatility, low costs and rapidity. Furthermore, with MSPD, it is possible to perform, on a small sample aliquot, extraction and clean up in a single step. In recent years, the greatest innovation in the MSPD technique has been the employment of unusual supporting materials (i.e. highly selective molecularly-imprinted polymers and the less specific multi-walled carbon nanotubes), although traditional sorbent and supports are still widely employed. Another novelty is the growing application of the multivariate statistical approach (experimental design) to optimize the extraction conditions.The purpose of this review is to present a rapid overview of MSPD applied to specific matrices and an update on the latest trends and innovations in the field since 2009. We pay particular attention to unusual matrices, new rapid clean-up techniques, and comparison with other sample-preparation methods.

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Research paper thumbnail of Evaluation of the atmospheric pressure photoionization source for the determination of benzidines and chloroanilines in water and industrial effluents by high performance liquid chromatography–tandem mass spectrometry

Talanta, 2007

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Research paper thumbnail of Flavonoid profile in soybeans by high-performance liquid chromatography/tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2007

Flavonoid profiling of a soybean sample has been performed by liquid chromatography/positive elec... more Flavonoid profiling of a soybean sample has been performed by liquid chromatography/positive electrospray tandem mass spectrometry (LC/ESI(+)-MS/MS) using a quadrupole-linear ion trap (QLIT) instrument with an information-dependent data acquisition (IDA) protocol that looped, in a single run, an enhanced MS scan and an enhanced product ion scan. As compromise between time and obtainable information, spectra acquisition was split into two distinct runs: 220-450 Th and 400-800 Th, respectively. The isoflavones daidzein and genistein were identified as aglycones, monoglycosides, diglycosides, triglycosides, acetylglycosides, malonylglycosides, malonyl diglycosides, and dimalonyl diglycosides, whereas glycitein triglycosides, acetylglycosides, and dimalonyl diglycosides were not detected. Also kaempferol di- and triglycosides, malonylglycosides and malonyl diglycosides, previously reported in soy leaves and pods, and four naringenin malonylglycosides were identified.

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Research paper thumbnail of Rapid-resolution liquid chromatography/mass spectrometry for determination and quantitation of polyphenols in grape berries

Rapid Communications in Mass Spectrometry, 2008

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Research paper thumbnail of Absolute quantification of cardiac troponin T by means of liquid chromatography/triple quadrupole tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2008

A liquid chromatography/tandem mass spectrometric method for absolute quantification of cardiac t... more A liquid chromatography/tandem mass spectrometric method for absolute quantification of cardiac troponin T (cTnT) in mouse heart tissue is presented. Even in such a complex biological sample, the multiple reaction monitoring acquisition mode allowed the selective and sensitive determination of a specific peptide, obtained by cTnT enzymatic digestion. The concentration of this cTnT-specific peptide was considered as a representation of the concentration of its parent protein. Quantification was carried out by means of the matrix-matched calibration curve, constructed by adding the synthetic standard of the target peptide and another synthetic structurally analogous peptide as internal standard. Method identification limit and method quantification limit were estimated as 60 and 110 ng of cTnT per mg of total extracted proteins, respectively. The developed label-free approach has been applied for the absolute quantitation of cTnT because of its diagnostic and prognostic value as cardiac disease marker. However, the method could be of general application, since it requires only the synthesis of two suitable peptides, a protein tryptic cleavage product and an internal standard.

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Research paper thumbnail of Determination of type B fumonisin mycotoxins in maize and maize-based products by liquid chromatography/tandem mass spectrometry using a QqQlinear ion trap mass spectrometer

Rapid Communications in Mass Spectrometry, 2005

A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for determining the ... more A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for determining the type B fumonisin mycotoxins in corn-based foodstuffs is described. Fumonisins FB1 and FB2 were extracted from a 1 g sample by homogenization with acetonitrile/water (75:25, v/v, 50 mmol/L formic acid, 25 mL final volume) and the extract was defatted on C18 phase. Volumes of 5 mL of crude extracts were cleaned up on Carbograph-4 cartridges. The final solution was analyzed by HPLC with electrospray ionization mass spectrometry in positive ion mode using multiple reaction monitoring with a QqQ linear ion trap mass spectrometer. Recoveries for spiked corn-based foodstuffs ranged from 91-105% (RSD% < or =8%), and method detection limits were < or =2 ng/g for FB1 and < or =1 ng/g for FB2. Two different spiking levels were tested (5000 and 100 ng/g for FB1, 1000 and 20 ng/g for FB2). Quantitation was achieved by an external calibration procedure using matrix-matched standards, with diclofenac added post-cleanup as internal standard for the LC/MS/MS analyses. Calibration curves showed linearity in the concentration range 0.005-5 ng/microL of final extract (0.992 < or = R2< or =0.995). Two other fumonisins, FB3 and FB4, were identified in naturally contaminated samples of corn meal using an information-dependent acquisition protocol that looped three experiments, including neutral loss scan, enhanced resolution scan, and enhanced product ion scan. FB3 and FB4 quantitation was estimated as peak area ratios relative to the FB2 response in view of the lack of both standards. This work also includes an application of the present LC/MS/MS method to some maize and maize-based product samples (corn meal, cornflakes and popcorn) collected from Italian stores. FB1 and FB2 contamination levels exceeding the European Union recommendation were found in 8 out of 15 corn meal samples.

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Research paper thumbnail of Identification and mass spectrometric characterization of glycosylated flavonoids inTriticum durum plants by high-performance liquid chromatography with tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2005

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Research paper thumbnail of Development and validation of a liquid chromatography/atmospheric pressure photoionization-tandem mass spectrometric method for the analysis of mycotoxins subjected to commission regulation (EC) No. 1881/2006 In cereals

Journal of Chromatography A, 2010

A sensitive and reliable liquid chromatography/photoionization (APPI) tandem mass spectrometry me... more A sensitive and reliable liquid chromatography/photoionization (APPI) tandem mass spectrometry method has been developed for determining nine selected mycotoxins in wheat and maize samples. The analytes were chosen on the basis of the mycotoxins under EU Commission Regulation (EC) No. 1881/2006, i.e., deoxynivalenol (DON), zearalenone (ZON), aflatoxins (AFs), and ochratoxin A (OTA), and considering the possibility of a near future

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Research paper thumbnail of Liquid chromatography/tandem mass spectrometric confirmatory method for determining aflatoxin M1 in cow milk

Journal of Chromatography A, 2006

A liquid chromatography/electrospray (ESI)–tandem mass spectrometric method for the measurement o... more A liquid chromatography/electrospray (ESI)–tandem mass spectrometric method for the measurement of aflatoxin M1 (AFM1) in milk is described. Milk sample after protein precipitation with acetone was cleaned-up with a Carbograph-4 cartridge. Performances of the ESI source were compared with those of the atmospheric pressure photoionization source (APPI). Although a method quantification limit (MQL) of 6ng/kg could be achieved operating with

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Research paper thumbnail of Reversed-phase high-performance liquid chromatographic determination of linear alkylbenzene sulphonates, nonylphenol polyethoxylates and their carboxylic biotransformation products

Journal of Chromatography A, 1993

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Research paper thumbnail of Ultra-high-performance liquid chromatography-tandem mass spectrometry for the analysis of free and conjugated natural estrogens in cow milk without deconjugation

Analytical and Bioanalytical Chemistry

A sensitive liquid chromatography/electrospray ionization-tandem mass spectrometry method for the... more A sensitive liquid chromatography/electrospray ionization-tandem mass spectrometry method for the determination of free and conjugated estrogens in cow milk is described. Milk samples were diluted with water and then extracted and cleaned up by solid-phase extraction using graphitized carbon black as adsorbent material, without any enzymatic cleavage, derivatization, and/or protein precipitation step. Two fractions were collected (free and conjugated estrogens) and analyzed separately. Mass spectrometry analysis was performed in negative ion mode using selected reaction monitoring acquisition mode. For all compounds, the coefficients of determination (R (2)) ranged between 0.9892 and 0.9997. Analytical recoveries were in the range of 86-109 % for free estrogens and 85-118 % for conjugates, with relative standard deviations below 10 %, and the method detection limit ranged between 3 and 80 ng L(-1). Finally, the developed method was used to determine the presence of free and conjugat...

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Research paper thumbnail of Liquid chromatography/tandem mass spectrometry determination of organophosphorus flame retardants and plasticizers in drinking and surface waters

Rapid Communications in Mass Spectrometry

A liquid chromatography/electrospray ionization tandem mass spectrometric method for analyzing or... more A liquid chromatography/electrospray ionization tandem mass spectrometric method for analyzing organophophorus flame retardants and plasticizers in drinking and environmental waters was developed. Five alkyl phosphates, three chlorinated alkyl phosphates, two aryl phosphate and triphenylphosphine oxide were selected for this study. These compounds were extracted from water samples by a hydrophilic polymeric solid-phase extraction cartridge. Accuracy and precision were evaluated analyzing 0.5 L of water samples spiked at concentrations of 10 and 100 ng/L for drinking water and at 300 and 1000 ng/L for river water. Except for trimethyl phosphate, analyte recoveries were better than 80%, and were not dependent on the type of aqueous matrix in which they were dissolved. At the spike levels considered, within-day precision was between 3 and 12% for tap water and between 4 and 14% for river water, and estimated method quantification limits ranged from 0.2 to 3.9 ng/L. A short survey condu...

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Research paper thumbnail of Peptidome characterization and bioactivity analysis of donkey milk

Journal of Proteomics, 2015

Donkey milk is an interesting commercial product for its nutritional values, which make it the mo... more Donkey milk is an interesting commercial product for its nutritional values, which make it the most suitable mammalian milk for human consumption, and for the bioactivity associated with it and derivative products. To further mine the characterization of donkey milk, an extensive peptidomic study was performed. Two peptide purification strategies were compared to remove native proteins and lipids and enrich the peptide fraction. In one case the whole protein content was precipitated by organic solvent using cold acetone. In the other one the precipitation of the most abundant milk proteins, caseins, was performed under acidic conditions by acetic acid at pH4.6, instead. The procedures were compared and proved to be partially complementary. Considered together they provided 1330 peptide identifications for donkey milk, mainly coming from the most abundant proteins in milk. The bioactivity of the isolated peptides was also investigated, both by angiotensin-converting-enzyme inhibitory and antioxidant activity assays and by bioinformatics, proving that the isolated peptides did have the tested biological activities. The rationale behind this study is that peptides in food matrices often play an important biological role and, despite the extensive study of the protein composition of different samples, they remain poorly characterized. In fact, in a typical shotgun proteomics study endogenous peptides are not properly characterized. In proteomics workflows one limiting point is the isolation process: if it is specific for the purification of proteins, it often comprises a precipitation step which aims at isolating pure protein pellets and remove unwonted interferent compounds. In this way endogenous peptides, which are not effectively precipitated as well as proteins, are removed too and not analyzed at the end of the process. Moreover, endogenous peptides do often originate from precursor proteins, but in phenomena which are independent of the shotgun digestion protocol, thus they can be obtained from cleavage specificities other than trypsin's, which is the main proteolytic enzyme employed in proteomic experiments. For this reason, in the end, database search will not be effective for identification of these peptides, thus the need to provide different workflows for peptide analysis. In the work presented in this paper this issue is considered for the first time for the analysis of the peptides isolated in donkey milk samples, which have been chosen for its nutritional interest. This study provides additional knowledge on this milk, already characterized by traditional proteomics studies and peptidomic studies after simulated digestion. This type of study is not just a description of the naturally occurring peptidome of a sample, but also represents a starting point to discover and characterize those naturally occurring peptides responsible for the observed bioactivities of biological samples, as in the case of donkey milk, which would remain uncharacterized by other approaches. In this paper an analytical protocol was described for the efficient isolation and purification of peptides in donkey milk, assessing the effect of the purification protocol on the final identifications. Purified peptide samples were also checked to empirically elucidate any ACE inhibitory or antioxidant activity. Finally, the peptidomic results were also further mined by a bioinformatic-driven approach for bioactive peptide identification in the donkey milk samples. In our opinion, the main strengths of this study are related to the improved analytical workflow (either as purification protocol comparison or analytical platform development) which provides a high number of identified peptides, for which the biological significance as potential bioactive peptides has also been investigated.

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Research paper thumbnail of Application of an innovative matrix solid-phase dispersion–solid-phase extraction–liquid chromatography–tandem mass spectrometry analytical methodology to the study of the metabolism of the estrogenic mycotoxin zearalenone in rainbow trout liver and muscular tissue

International Journal of Environmental Analytical Chemistry, 2004

The metabolic fate of the estrogenic mycotoxin zearalenone in rainbow trout is presently unknown.... more The metabolic fate of the estrogenic mycotoxin zearalenone in rainbow trout is presently unknown. In this study, the tissue concentration of zearalenone and its principal metabolites (α-zearalenol and β-zearalenol) was determined. A known amount of zearalenone was administered as a single bolus to ten fish, and the biological tissue concentration was determined at various times following administration. The analytes were

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Research paper thumbnail of Protein Profile of Mature Soybean Seeds and Prepared Soybean Milk

Journal of Agricultural and Food Chemistry, 2014

The soybean (Glycine max (L.) Merrill) is economically the most important bean in the world, prov... more The soybean (Glycine max (L.) Merrill) is economically the most important bean in the world, providing a wide range of vegetable proteins. Soybean milk is a colloidal solution obtained as water extract from swelled and ground soybean seeds. Soybean proteins represent about 35-40% on a dry weight basis and they are receiving increasing attention with respect to their health effects. However, the soybean is a well-recognized allergenic food, and therefore, it is urgent to define its protein components responsible for the allergenicity in order to develop hypoallergenic soybean products for sensitive people. The main aim of this work was the characterization of seed and milk soybean proteome and their comparison in terms of protein content and specific proteins. Using a shotgun proteomics approach, 243 nonredundant proteins were identified in mature soybean seeds.

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Research paper thumbnail of Multiresidue determination of UV filters in water samples by solid-phase extraction and liquid chromatography with tandem mass spectrometry analysis

Journal of Separation Science, 2014

UV filters, contained in sunscreens and other cosmetic products, as well as in some plastics and ... more UV filters, contained in sunscreens and other cosmetic products, as well as in some plastics and industrial products, are nowadays considered contaminants of emerging concern because their widespread and increasing use has lead to their presence in the environment. Furthermore, some UV filters are suspected to have endocrine disruption activity. In the present work, we developed an analytical method based on liquid chromatography with tandem mass spectrometry for the determination of UV filters in tap and lake waters. Sixteen UV filters were extracted from water samples by solid-phase extraction employing graphitized carbon black as adsorbent material. Handling 200 mL of water sample, satisfactory recoveries were obtained for almost all the analytes. The limits of detection and quantification of the method were comparable to those reported in other works, and ranged between 0.7-3.5 and 1.9-11.8 ng/L, respectively; however in our case the number of investigated compounds was larger. The major encountered problem in method development was to identify the background contamination sources and reduce their contribution. UV filters were not detected in tap water samples, whereas the analyses conducted on samples collected from three different lakes showed that the swimming areas are most subject to UV filter contamination.

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Research paper thumbnail of Development of an analytical strategy for the identification of potential bioactive peptides generated by in vitro tryptic digestion of fish muscle proteins

In the last years, food proteins and peptides are attracting great attention because of the emerg... more In the last years, food proteins and peptides are attracting great attention because of the emergence of a new field, that of food-derived bioactive peptides. This paper presents a comparison and evaluation of four different experiments for the identification of sarcoplasmic and myofibrillar fish peptides. This study is aimed at the development of a simple and fast method for the identification of peptides that could arise from fish meat if trypsin was the only digestive enzyme acting on fish meat proteins. In particular, we tested the use of ultrafiltration membranes with a molecular weight cutoff of 3,000 Da. Data analysis has shown that the experiment in which there is neither precipitation nor an ultrafiltration step performed better and allowed the identification of a larger number of peptides and potential antimicrobial peptides (AMPs); this workflow provided 473 and 398 total identified peptides and 44 and 18 AMPs for sarcoplasmic and myofibrillar extracts, respectively. This protocol is found to be faster and more straightforward than the other three tested workflows. The developed strategy could be also useful for other food matrices and could provide information about food quality and safety control.

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Research paper thumbnail of Effect of membrane charge density on the protein corona of cationic liposomes: Interplay between cationic charge and surface area

Applied Physics Letters, 2011

Here we investigate the effect of membrane charge density on the protein corona that forms upon e... more Here we investigate the effect of membrane charge density on the protein corona that forms upon exposure of cationic liposomes to human plasma. We show that the protein corona around cationic liposomes is not uniquely defined by membrane charge density, but it is deeply affected by both the overall cationic charge and the available surface area. Remarkably, changes in protein

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Research paper thumbnail of Comparative analysis of metabolic proteome variation in ascorbate-primed and unprimed wheat seeds during germination under salt stress

Journal of Proteomics, 2014

Seed priming with ascorbic acid improves salt tolerance in durum wheat. For understanding the pot... more Seed priming with ascorbic acid improves salt tolerance in durum wheat. For understanding the potential mechanisms underlying this priming effect a gel-free shotgun proteomic analysis was performed comparing unprimed to ascorbate-primed wheat seed during germination under saline and non-saline conditions. Since seed germination is the result of interplay or cross-talk between embryo and embryo-surrounding tissues, we studied the variation of metabolic proteome in both tissues separately. 167 of 697 identified and 69 of 471 identified proteins increase or decrease in abundance significantly in response to priming and/or salinity compared to untreated, unstressed control in embryo and embryo-surrounding tissues, respectively. In untreated wheat embryo salt stress was accompanied by change in 129 proteins, most of which are belonging to metabolism, energy, disease/defense, protein destination and storage categories. Ascorbate pretreatment prevents and counteracts the effects of salinity upon most of these proteins and changes specifically the abundance of 35 others proteins, most of which are involved in metabolism, protein destination and storage categories. Hierarchical clustering analysis revealed three and two major clusters of protein expression in embryo and embryo-surrounding tissues, respectively. This study opens promising new avenues to understand priming-induced salt tolerance in plants. To clearly understand how ascorbate-priming enhance the salt tolerance of durum wheat during germination, we performed for the first time a comparative shotgun proteomic analysis between unprimed and ascorbate-primed wheat seeds during germination under saline and non-saline conditions. Furthermore, since seed germination is the result of interplay or cross-talk between embryo and embryo-surrounding tissues we analyzed the variation of metabolic proteome in both tissues separately. 1168 proteins exhibiting greater molecular weight diversity (ranging from 5 to 258kDa) were identified. Among them, 167 and 69 proteins were increased or decreased in abundance significantly by priming and/or salinity as compared to control, in embryo and embryo-surrounding tissues respectively. Ascorbate pretreatment alleviates the effects of salinity upon most of these proteins, particularly those involved in metabolism, energy, disease/defense, protein destination and storage functions. Hierarchical clustering analysis revealed three and two major clusters of protein accumulation in embryo and embryo-surrounding tissues, respectively. These results may provide new avenues for understanding and advancing priming-induced salt tolerance in crop plants.

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Research paper thumbnail of Recent trends in matrix solid-phase dispersion

TrAC Trends in Analytical Chemistry, 2013

ABSTRACT The key factors for the success of matrix solid-phase dispersion (MSPD) are its feasibil... more ABSTRACT The key factors for the success of matrix solid-phase dispersion (MSPD) are its feasibility, flexibility, versatility, low costs and rapidity. Furthermore, with MSPD, it is possible to perform, on a small sample aliquot, extraction and clean up in a single step. In recent years, the greatest innovation in the MSPD technique has been the employment of unusual supporting materials (i.e. highly selective molecularly-imprinted polymers and the less specific multi-walled carbon nanotubes), although traditional sorbent and supports are still widely employed. Another novelty is the growing application of the multivariate statistical approach (experimental design) to optimize the extraction conditions.The purpose of this review is to present a rapid overview of MSPD applied to specific matrices and an update on the latest trends and innovations in the field since 2009. We pay particular attention to unusual matrices, new rapid clean-up techniques, and comparison with other sample-preparation methods.

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Research paper thumbnail of Evaluation of the atmospheric pressure photoionization source for the determination of benzidines and chloroanilines in water and industrial effluents by high performance liquid chromatography–tandem mass spectrometry

Talanta, 2007

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Research paper thumbnail of Flavonoid profile in soybeans by high-performance liquid chromatography/tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2007

Flavonoid profiling of a soybean sample has been performed by liquid chromatography/positive elec... more Flavonoid profiling of a soybean sample has been performed by liquid chromatography/positive electrospray tandem mass spectrometry (LC/ESI(+)-MS/MS) using a quadrupole-linear ion trap (QLIT) instrument with an information-dependent data acquisition (IDA) protocol that looped, in a single run, an enhanced MS scan and an enhanced product ion scan. As compromise between time and obtainable information, spectra acquisition was split into two distinct runs: 220-450 Th and 400-800 Th, respectively. The isoflavones daidzein and genistein were identified as aglycones, monoglycosides, diglycosides, triglycosides, acetylglycosides, malonylglycosides, malonyl diglycosides, and dimalonyl diglycosides, whereas glycitein triglycosides, acetylglycosides, and dimalonyl diglycosides were not detected. Also kaempferol di- and triglycosides, malonylglycosides and malonyl diglycosides, previously reported in soy leaves and pods, and four naringenin malonylglycosides were identified.

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Research paper thumbnail of Rapid-resolution liquid chromatography/mass spectrometry for determination and quantitation of polyphenols in grape berries

Rapid Communications in Mass Spectrometry, 2008

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Research paper thumbnail of Absolute quantification of cardiac troponin T by means of liquid chromatography/triple quadrupole tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2008

A liquid chromatography/tandem mass spectrometric method for absolute quantification of cardiac t... more A liquid chromatography/tandem mass spectrometric method for absolute quantification of cardiac troponin T (cTnT) in mouse heart tissue is presented. Even in such a complex biological sample, the multiple reaction monitoring acquisition mode allowed the selective and sensitive determination of a specific peptide, obtained by cTnT enzymatic digestion. The concentration of this cTnT-specific peptide was considered as a representation of the concentration of its parent protein. Quantification was carried out by means of the matrix-matched calibration curve, constructed by adding the synthetic standard of the target peptide and another synthetic structurally analogous peptide as internal standard. Method identification limit and method quantification limit were estimated as 60 and 110 ng of cTnT per mg of total extracted proteins, respectively. The developed label-free approach has been applied for the absolute quantitation of cTnT because of its diagnostic and prognostic value as cardiac disease marker. However, the method could be of general application, since it requires only the synthesis of two suitable peptides, a protein tryptic cleavage product and an internal standard.

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Research paper thumbnail of Determination of type B fumonisin mycotoxins in maize and maize-based products by liquid chromatography/tandem mass spectrometry using a QqQlinear ion trap mass spectrometer

Rapid Communications in Mass Spectrometry, 2005

A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for determining the ... more A sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for determining the type B fumonisin mycotoxins in corn-based foodstuffs is described. Fumonisins FB1 and FB2 were extracted from a 1 g sample by homogenization with acetonitrile/water (75:25, v/v, 50 mmol/L formic acid, 25 mL final volume) and the extract was defatted on C18 phase. Volumes of 5 mL of crude extracts were cleaned up on Carbograph-4 cartridges. The final solution was analyzed by HPLC with electrospray ionization mass spectrometry in positive ion mode using multiple reaction monitoring with a QqQ linear ion trap mass spectrometer. Recoveries for spiked corn-based foodstuffs ranged from 91-105% (RSD% < or =8%), and method detection limits were < or =2 ng/g for FB1 and < or =1 ng/g for FB2. Two different spiking levels were tested (5000 and 100 ng/g for FB1, 1000 and 20 ng/g for FB2). Quantitation was achieved by an external calibration procedure using matrix-matched standards, with diclofenac added post-cleanup as internal standard for the LC/MS/MS analyses. Calibration curves showed linearity in the concentration range 0.005-5 ng/microL of final extract (0.992 < or = R2< or =0.995). Two other fumonisins, FB3 and FB4, were identified in naturally contaminated samples of corn meal using an information-dependent acquisition protocol that looped three experiments, including neutral loss scan, enhanced resolution scan, and enhanced product ion scan. FB3 and FB4 quantitation was estimated as peak area ratios relative to the FB2 response in view of the lack of both standards. This work also includes an application of the present LC/MS/MS method to some maize and maize-based product samples (corn meal, cornflakes and popcorn) collected from Italian stores. FB1 and FB2 contamination levels exceeding the European Union recommendation were found in 8 out of 15 corn meal samples.

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Research paper thumbnail of Identification and mass spectrometric characterization of glycosylated flavonoids inTriticum durum plants by high-performance liquid chromatography with tandem mass spectrometry

Rapid Communications in Mass Spectrometry, 2005

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