A synthesis of representative monohydroxy derivatives of valinomycin (VLM) was achieved under mild conditions by direct hydroxylation at the side chains of the macrocyclic substrate using dioxiranes. Results demonstrate that the powerful methyl(trifluoromethyl)dioxirane 1b should be the reagent of choice to carry out these key transformations. Thus, a mixture of compounds derived from the direct dioxirane attack at the β-(CH3)2CH alkyl chain of one Hyi residue (compound 3a) or of one Val moiety (compounds 3b and 3c) could be obtained. Following convenient mixture separation, each of the new oxyfunctionalized macrocycles became completely characterized.

The effectiveness of a novel binary matrix composed of 1,8-bis(dimethylamino)naphthalene (DMAN;proton sponge) and 9-aminoacridine (9AA) for the direct lipid analysis of whole bacterial cells by matrixassisted laser desorption ionization mass spectrometry (MALDI MS) is demonstrated. Deprotonated ana-lyte signals nearly free of matrix-related ions were observed in negative ion mode. The effect of themost important factors (laser energy, pulse voltage, DMAN/9AA ratio, analyte/matrix ratio) was investi-gated using a Box–Behnken response surface design followed by multi-response optimization in order tosimultaneously maximize signal-to-noise (S/N) ratio and resolution. The chemical surface compositionof single or mixed matrices was explored by X-ray photoelectron spectroscopy (XPS). Moreover, XPSimaging was used to map the spatial distribution of a model phospholipid in single or binary matrices.The DMAN/9AA binary matrix was then successfully applied to the analysis of intact Gram positive(Lactobacillus sanfranciscensis) or Gram negative (Escherichia coli) microorganisms. About fifty majormembrane components (free fatty acids, mono-, di- and tri-glycerides, phospholipids, glycolipids andcardiolipins) were quickly and easily detected over a mass range spanning from ca. 200 to ca. 1600 m/z.Moreover, mass spectra with improved S/N ratio (compared to single matrices), reduced chemical noiseand no formation of matrix-clusters were invariably obtained demonstrating the potential of this binarymatrix to improve sensitivity.

Nowadays, matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry represents an emerging and versatile tool for analysis of lipids. However, direct (i.e., with no previous separation of lipid classes) analysis of crude extracts containing a complex mixture of lipids (a problem typically encountered in shotgun lipidomics) is still a quite challenging task using a conventional MALDI matrix such as 2,5-dihydroxybenzoic acid (DHB). Indeed, in the presence of phospholipids containing quaternary ammonium groups, such as phosphatidylcholines and sphingomyelins, strong ionization-suppression effects are experienced especially in positive ion mode. To overcome this limitation, lumazine (1H-pteridine-2,4-dione) was evaluated as an alternative matrix. Lumazine in the solid state showed an absorption maximum at 350 nm, ionizes/desorbs without appreciable decomposition and extensive cluster formation, and can be used in both ion modes. In positive ion mode, the main species were M(center dot+) and 2M(center dot+) radical cations and cationized species ([M+H](+), [M+Na](+), [M+2Na+2Li-3H](+)). In negative ion mode, the main signals observed were the deprotonated molecular ion and the radical anion. The signal-to-noise ratio for phosphatidylglycerols and phosphatidylethanol-amines using lumazine was almost 1 order of magnitude higher than that observed for DHB. Lumazine was successfully used for MALDI analysis (positive and negative ion modes) of crude lipid extracts of milk, soymilk, and hen egg, where phosphatidylethanolamines, phosphatidylserines, and phosphatidylinositols could additionally be detected.

A simple protocol, based on Bligh–Dyer (BD) extraction followed by MALDI-TOF-MS analysis, for fast identification of paint binders in single microsamples is proposed. For the first time it is demonstrated that the BD method is effective for the simultaneous extraction of lipids and proteins from complex, and atypical matrices, such as pigmented paint layers. The protocol makes use of an alternative denaturing anionic detergent (RapiGestTM) in order to improve efficiency of protein digestion and purification step. Detection of various lipid classes, such as triacylglycerols (TAGs) and phospholipids (PLs), and their oxidation by-products was accomplished, whereas proteins could be identified by peptide mass fingerprinting. The effect of pigments on ageing of lipids and proteins was also investigated. Finally, the proposed protocol was successfully applied to the study of a late-15th century Italian panel painting allowing the identification of various proteinaceous and lipid sections in organic binders, such as egg yolk, egg white, animal glue, casein, and drying oil. ©

A boronic analogue of the archetype matrix a-cyano-4-hydroxycinnamic acid (CHCA) has been synthesised providing a new ‘‘reactive matrix’’ that possesses molecular recognition properties. This matrix selectively recognizes vic-diols, a-hydroxyacids, aminols and first allowed the detection of anions as fluoride (unaffordable by usual matrices).

The oil polar fraction may have a great potential for the characterization of vegetable oils and for the individuation of adulterations. In particular, adulteration of extra-virgin olive oil (EVOO) with hazelnut oil (HO) is one of the most difficult ones to detect due to the similar composition as regards triacylglycerol, total sterol and fatty acid profile. A new micro-solid phase extraction (μ-SPE) procedure based on hydrophilic liquid chromatography (HILIC) micro-columns was developed for the selective extraction and enrichment of polar compounds from EVOO and HO beforematrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-ToF-MS) analysis. The method permits a simple and fast qualitative analysis of the polar fraction of the oils under study; furthermore, somepeaks (such as them/z ions 496.39, 520.46 and 522.47) were found tobe present only inHO, indicating that they could be diagnostic for the presence ofHOin EVOO. In order to verify the potential of the method for the individuation of this adulteration,EVOOwas progressively adulteratedwith variable quantities ofHO, subjected to the HILIC enrichment and finally to MALDI-ToF-MS analysis; the detection of adulteration was possible up to the level of 5%. Eventually, diagnostic polar compoundswere identified as lysophosphatidylcholine (LPC) (16 : 0/0 : 0), LPC (18 : 2/0 : 0), LPC (18 : 1/0 : 0) by means of capillary liquid chromatography-electrospray ionization-quadrupole-ToF-MS (CapLC-ESI-Q-ToF-MS) analysis.

A matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based approach was applied for the detection of various lipid classes, such as triacylglycerols (TAGs) and phospholipids (PLs), and their oxidation by-products in extracts of small (50–100 μg) samples obtained from painted artworks. Ageing of test specimens under various conditions, including the presence of different pigments, was preliminarily investigated. During ageing, the TAGs and PLs content decreased, whereas the amount of diglycerides, short-chain oxidative products arising from TAGs and PLs, and oxidized TAGs and PLs components increased. The examination of a series of model paint samples gave a clear indication that specific ions produced by oxidative cleavage of PLs and/or TAGs may be used as markers for egg and drying oil-based binders. Their elemental composition and hypothetical structure are also tentatively proposed. Moreover, the simultaneous presence of egg and oil binders can be easily and unambiguously ascertained through the simultaneous occurrence of the relevant specific markers. The potential of the proposed approach was demonstrated for the first time by the analysis of real samples from a polyptych of Bartolomeo Vivarini (fifteenth century) and a “French school” canvas painting (seventeenth century).

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 and MALDI-TOF of nuclear extracts show that Avp triggers Avpr1α localization to the nucleus. Finally, a specific Avpr2 inhibitor, tolvaptan, does not affect bone formation or bone mass, suggesting that Avpr2, which primarily functions in the kidney, does not have a significant role in bone remodeling.

A method of direct lipid analysis by matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry (MS) in intact membranes, without prior extraction/separation steps, is described. Here, we demonstrate the efficacy of a strong base, 1,8‐bis(dimethylamino)naphthalene(DMAN; proton sponge), as a novel matrix for MALDI‐time‐of‐flight (TOF) MS analysis of whole cell bacteria. Initially, individual acidic low‐molecular‐weight analytes such as standard free fatty acids and phospholipids were analyzed using DMAN asmatrix. Clear negative‐modeMALDI‐TOFMS spectra of all analytes show only deprotonated analyte signals at a low picomole limit of detection with the complete absence of matrix‐related signals. These results indicate that DMAN represents a suitable matrix for MALDI‐TOF MS analysis of mixtures of complex lipids as the intact membranes of microorganisms. DMAN was successfully applied to the analysis of Lactobacillus sanfranciscensis and L. plantarum microorganisms. Different components were sensitively detected in a single spot, including 16:0, 18:2, 18:3, and 21:0 free acids, glycolipids, phosphatidylglycerols (PGs) and cardiolipins. This method might be of general application, offering the advantage of quickly gaining information about lipid components of other Gram‐positive bacterial membranes.

A simple method based on hydrophilic liquid chromatography (HILIC) and electrospray mass spectrometry (ESI-MS) for the detection of monoethanolamine (MEA) degradation products in CO2 post-combustion capture plants has been developed. MEA by-products determination has traditionally been difficult due to analytical separation problems. Even in recent sophisticated methods, this difficulty remains as the major issue often resulting in time-consuming sample preparations. In this work, we have collected samples directly from a real pilot plant and analyzed them, for the first time, by using matrix assisted laser desorption ionization (MALDI)-MS or ESIMS without any separation, both in positive and negative ionization modes. Alternatively, a previous liquid chromatography (LC) run was performed before ESI-MS; traditional reverse phase separation and HILIC were compared. Our results indicated that HILIC separation using an amino modified column, coupled to ESI-MS or ESI-MS/MS measurements, is the suitable method for identifying as many degradation products. Moreover, some plausible degradation mechanisms are proposed to explain some peaks in the spectra. The present work is intended as a preliminary study aiming to show the usefulness of these alternative techniques for this kind of investigations.

In the dairy industry one of the most common frauds is mixing high-value milk (sheep’s and goats’) with milk of lower value (cows’). This illegal practice has commercial, ethical, and serious sanitary consequences because consumers can be exposed to hidden allergens contained in the undeclared cows’ milk. Here, we investigated the possibility of using matrix-assisted laser-desorption/ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) as a rapid, sensitive, and accurate technique for detection of milk adulteration by analysis of phospholipid profiles. Lipid extracts of pure raw milk, commercial milk, and binary mixtures of cows’ and goats’ milk and cows’ and sheep’s milk (the concentrations of each milk varied from 0 % to 50 %) were analyzed with α-cyano-4-chlorocinnamic acid as matrix. The abundance ratio of the ions at m/z 703 and m/z 706 was found to be species-correlated and was used as marker of cows’ milk in sheep’s and goats’ milk. Furthermore, the procedure could potentially be applied to cheese samples, because peaks at m/z 703 and 706 were also found in several commercial cheese samples. This approach proved to be an efficient, rapid, and inexpensive method of detecting milk fraud.

The major modifications induced by thermal treatment of whey proteins R-lactalbumin (R-La) and β-lactoglobulin (β-Lg) in a model system mimicking lactose-free milk (L- sugar mix) were investigated by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The analysis of the intact R-La revealed species with up to 7 and 14 adducts from lactose and sugar mix, respectively, whereas for β-Lg 3 and up to 5 sugar moieties were observed in the case of lactose and sugar mix experiments, respectively. A partial enzymatic hydrolysis with endoproteinase AspN prior to mass spectrometric analysis allowed the detection of further modifications and their localization in the amino acid sequence. Using R-cyano-4-chlorocinnamic acid as MALDI matrix, it could be shown that heating R-La and β-Lg with glucose or galactose led to the modification of lysine residues that are not glycated by lactose. The higher glycation degree of whey proteins in a lactose-free milk system relative to normal milk with lactose reflects the higher reactivity of monosaccharides compared to the parent disaccharide. Finally, the analysis of the whey extract of a commercial lactose-free milk sample revealed that the two whey proteins were present as three main forms (native, single, and double hexose adducts).

The adulteration of extra virgin olive oil (EVOO) with hazelnut oil (HO) is frequent and constitutes a seri- ous concern both for oil suppliers and consumers. The high degree of similarity between the two oils as regards triacylglycerol, total sterol and fatty acid profile, complicates the detection of low percentages of HO in EVOO. However, phospholipids (PLs) are usually present in seed oils at a concentration range of 10– 20 g/kg, while the amounts of PLs in VOOs are 300–400 times lower. Thus, in this work a sample pretreat- ment procedure focused towards the selective PLs extraction was developed; the Bligh–Dyer extraction procedure was modified introducing the ionic liquid resulting from the combination of TBA (tributyl- amine) and CHCA (a-Cyano-4-hydroxycinnamic acid) as extraction solvent. The selective extraction and enrichment of phospholipids from EVOO and HO samples was then achieved. The relevant extracts were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI- TOF-MS) using the same ionic liquid TBA-CHCA as MALDI matrix, that was found to be very suitable for PLs analysis. In fact, a remarkable increase of the phospholipids signals, with a simultaneous decrease of those relevant to triacylglycerols and diacylglycerols, was observed in the relevant mass spectra. The applicability of the whole method to the individuation of the presence of HO in EVOO was demonstrated by the analysis of EVOO samples progressively adulterated with variable quantities of HO, that was still detectable at a 1% contamination level.

Escherichia coli (E. coli) is one of the most important foodborne pathogens to the food industry responsible for diseases as bloody diarrhea, hemorrhagic colitis and life-threatening hemolytic–uremic syndrome. For controlling and eliminating E. coli, metal nano- antimicrobials (NAMs) are frequently used as bioactive systems for applications in food treatments. Most NAMs provide controlled release of metal ions, eventually slowing down or completely inhibiting the growth of undesired microorganisms. Nonetheless, their antimicrobial action is not totally unraveled and is strongly dependent on metal properties and environmental conditions. In this work, we propose the use of matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry as a powerful tool for direct, time efficient, plausible identification of the cell membrane damage in bacterial strains exposed to copper-based antimicrobial agents, such as soluble salts (chosen as simplified AM material) and copper nanoparticles. E. coli ATCC 25922 strain was selected as ‘training bacterium’ to set up some critical experimental parameters (i.e. cell concentration, selection of the MALDI matrix, optimal solvent composition, sample preparation method) for the MS analyses. The resulting procedure was then used to attain both protein and lipid fingerprints from E. coli after exposure to different loadings of Cu salts and NPs. Inter- estingly, bacteria exposed to copper showed over-expression of copper binding proteins and degradation of lipids when treated with soluble salt. These findings were completed with other investigations, such as microbiological experiments.

The Crucifix panel painting in the Santa Maria a Mare church on the Isle of St. Nicholas (Isole Tremiti, Italy), painted on both sides, was executed between the late 12th century and the early 13th century and several times restored in the following centuries. The precious artefact was studied by various complementary analytical techniques in order to characterize the original medieval painting technique and the subsequently applied restoration materials. Optical microscopy (OM), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDS), micro-Raman spectroscopy, pyrolysis-gas chromatography/mass spectrometry (Py-GC/ MS), and Matrix Assisted Laser Desorption Ionisation–Time of Flight–Mass Spectrometry (MALDI-TOF-MS) were applied on various samples taken from significant parts of the painting. The compositional data were used for a correct planning of the recent restoration treatments and as a support for the historical-artistic study of the painting. The results obtained confirm that both paintings—recto and verso—were realized by following the 13th century Italian painting tradition. Egg-based paint layerswere applied on a gypsum/animal glue ground. Various pigments could be identified amongwhich the precious lapis lazuli. Interestingly, thewater-gilding of the recto was performed without the use of a bole layer. Pinaceae resin as well as acrylic resins were found.

In this study, the simultaneous identification of lipids and proteins is achieved by matrix assisted laser desorption ionization - mass spectrometry (MALDI-MS) analysis after direct on-plate processing of micro-samples supported on graphite. Taking advantages of large surface area and thermal conductivity, graphite provided an ideal substrate for on-plate proteolysis and lipid extraction without interfering with MALDI analysis. The new protocol was first developed on simple standards as bovine serum albumin to set up the best experimental conditions for proteolysis. Then, its feasibility and performance were demonstrated by processing more complex samples as milk and egg powder for simultaneous lipid/protein analysis. By such a protocol, proteins could be efficiently digested on-plate within 15 minutes, with sequence coverages comparable to those obtained by conventional overnight in-solution digestion. As an additional feature, detection of hydrophilic phosphorylated peptides could be achieved without any further enrichment procedure. Furthermore, lipids could be identified in the same analysis without any additional treatment/processing step. The present strategy is simple and efficient, of large applicability, offering great promise for high-throughput proteo-lipidomics in very small samples since no conventional solvent extraction is required reducing sample loss.

Protein analysis in biological fluids, such as urine, by means of mass spectrometry (MS) still suffers for insufficient standardization in protocols for sample collection, storage and preparation. In this work, the influence of these variables on healthy donors human urine protein profiling performed by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was studied. A screening of various urine sample pre-treatment procedures and different sample deposition approaches on the MALDI target was performed. The influence of urine samples storage time and temperature on spectral profiles was evaluated by means of principal component analysis (PCA). The whole optimized procedure was eventually applied to the MALDI-TOF-MS analysis of human urine samples taken from prostate cancer patients. The best results in terms of detected ions number and abundance in the MS spectra were obtained by using home-made microcolumns packed with hydrophilic-lipophilic balance (HLB) resin as sample pre-treatment method; this procedure was also less expensive and suitable for high throughput analyses. Afterwards, the spin coating approach for sample deposition on the MALDI target plate was optimized, obtaining homogenous and reproducible spots. Then, PCA indicated that low storage temperatures of acidified and centrifuged samples, together with short handling time, allowed to obtain reproducible profiles without artifacts contribution due to experimental conditions. Finally, interesting differences were found by comparing the MALDI-TOF-MS protein profiles of pooled urine samples of healthy donors and prostate cancer patients. The results showed that analytical and pre-analytical variables are crucial for the success of urine analysis, to obtain meaningful and reproducible data, even if the intra-patient variability is very difficult to avoid. It has been proven how pooled urine samples can be an interesting way to make easier the comparison between healthy and pathological samples and to individuate possible differences in the protein expression between the two sets of samples.

We report that oxytocin (Oxt) receptors (Oxtrs), on stimulation by the ligand Oxt, translocate into the nucleus of osteoblasts, implicating this process in the action of Oxt on osteoblast maturation. Sequential immunocytochemistry of intact cells or isolated nucleoplasts stripped of the outer nuclear membrane showed progressive nuclear localization of the Oxtr; this nuclear translocation was confirmed by monitoring the movement of Oxtr-EGFP as well as by immunogold labeling. Nuclear Oxtr localization was conclusively shown by Western immunoblotting and MS of nuclear lysate proteins. We found that the passage of Oxtrs into the nucleus was facilitated by successive interactions with β-arrestins (Arrbs), the small GTPase Rab5, importin-β (Kpnb1), and transportin-1 (Tnpo1). siRNA-mediated knockdown of Arrb1, Arrb2, or Tnpo1 abrogated Oxt-induced expression of the osteoblast differentiation genes osterix (Sp7), Atf4, bone sialoprotein (Ibsp), and osteocalcin (Bglap) without affecting Erk phosphorylation. Likewise and again, without affecting pErk, inhibiting Arrb recruitment by mutating Ser rich clusters of the nuclear localization signal to Ala abolished nuclear import and Oxtr-induced gene expression. These studies define a previously unidentified mechanism for Oxtr action on bone and open possibilities for direct transcriptional modulation by nuclear G protein-coupled receptors.

An efficient N-selective alkylation of primary aromatic amines in molten quaternary ammonium salts, as the solvent, under relatively mild and base-free conditions is presented. On the basis of the Kamlet–Taft parameters and the nucleophilicity of the IL (ionic liquid) anions, the influence of the ionic liquid was evaluated. This protocol was validated on a larger multigram scale and with the syntheses of bioactive heterocycles (e.g., 1,4-benzothiazine and quinoxalines) and new efficient MALDI matrixes.

A solid phase microextraction—liquid chromatography with ultraviolet detection (SPME-LC-UV) method for the determination of the antimicrobial agent chloramphenicol was developed. The performances of three commercially available fibers were compared; the Carbowax/TPR-100 was found to provide the most efficient extraction. All the aspects influencing the fiber adsorption (extraction time, temperature, pH, salt addition) and desorption (desorption and injection time, desorption solvent mixture composition) of the analyte were investigated. The method was eventually applied to the determination of the drug in both biological (urine) and environmental (tap and sea water) samples. The optimized procedure required a simple sample pretreatment, isocratic elution, and provided enough sensitivity for the analyte determination in the considered samples. The investigated linear ranges were 37–1000 ng/ml (urine), 0.1–10 ng/ml (tap water), 0.3–30 ng/ml (sea water). Within-day and between-days RSD% ranged between 5.5–6.2 and 8.7–9.0 (urine), 5.1–6.0 and 8.4–8.8 (tap water), 5.4–5.7 and 8.6–8.9 (sea water). Estimated LOD and LOQ were 37 and 95 ng/ml (urine), 0.1 and 0.3 ng/ml (tap water), 0.3 and 0.7 ng/ml (sea water).

Oleuropein (Ole) has been claimed to mitigate cisplatin (CP) induced acute injury in kidney and liver of mice. In-vitro reactivity of hydrated CP species with Ole, and Ole metabolite hydroxytyrosol (HT) is of great interest as the preliminary step for gathering in-vivo information on the possible physiological role of the Ole/HT-cis-diammineplatinum (II) (Ole/HT-cis-DAP) conjugate.

Low temperature treatments commonly applied to seafood products have been shown to influence their phospholipid (PL) profile through enzymatic hydrolysis. In the present study, the generation of lysophospholipids (LPL) resulting from this process was systematically investigated for selected, commercially relevant seafood products, namely oysters, clams, octopuses, and shrimps. These products were subjected to thermal treatments like refrigeration or freezing after being purchased as fresh, defrozen, or frozen products depending on the case. The coupling between hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization with high resolution/accuracy Fourier transform mass spectrometry (ESI-FTMS) was exploited to evaluate the PL profile of the cited products, especially the incidence of LPL related to the two main PL classes of seafood products-phosphatidylcholines (PC) and phosphatidylethanolamines (PE)-in the lipid extracts. The lyso forms of PE (LPE) were found to be generally more sensitive than those of PC (LPC) to thermal treatments, usually exhibiting a significant increase upon prolonged refrigeration at 4 °C in all types of investigated products except European flat oysters. Moreover, the distinction between fresh and frozen or defrozen products could be achieved in the case of octopuses and shrimps, respectively.

The processes of lipids oxidation represent a great concern for the consumer health because they are one of the major causes of quality deterioration in fat-containing products. One of the most effective methods of delaying lipid oxidation consists in incorporating antioxidants. The present investigation describes the formulation of chitosan and novel glycol chitosan nanoparticles (NPs) loaded with α-Tocopherol (αToc-NPs). The obtained NPs were characterized by various techniques, such as particle size (showing mean diameters in the range 335-503 nm) and zeta potential measurements, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. The NPs were, then, added in the preparation of oil-in-water simple emulsion both to make the lipophilic αToc available in an aqueous medium and to prevent emulsion oxidation. For this purpose, a new highly sensitive, simple and solvent-free method based on a solid phase microextraction (SPME) coupled to gas chromatography mass spectrometry was developed for the determination of αToc in aqueous medium. All the parameters influencing SPME, including fiber coating, time and temperature extraction, pH, ionic strength and desorption conditions, have been carefully screened. The method was successfully applied to the determination of vitamin in the αToc-NPs and its release from NPs-enriched simple emulsion formulations. SPME provided high recovery yields and the limits of detection and of quantification in emulsion were 0.1 and 0.5 μg/mg, respectively. The precision of the method has been also estimated. The delay of the lipid oxidation by the proposed formulations has been evaluated exploiting the Kreis test on αToc-NPs-enriched emulsions.