Cyclopropane fatty acids (CPFAs) are unusual fatty acids of microbial origin, recently detected in milk and dairy products. CPFAs have been demonstrated to be interesting molecular markers for authentication of dairy products obtained without ensiled feeds. Moreover, they can also be recognized as a new secondary component of human diet. Information is lacking on the presence of cyclic fatty acids in other food sources. Cyclopropane fatty acids have been detected by GC-MS analysis in cheese and other animal fats in concentration ranging from 200 to 1000 mg/kg fat, but in some cases, the complex fatty acid profile and the possible presence of interfering peaks make the separation not straightforward and the quantification uneasy. Therefore, a new reliable¹H NMR method was developed to detect and measure CPFA content in different foods of animal origin, based on the detection of the characteristic signals of cyclopropane ring. The¹H NMR (600 MHz) method showed detection limits comparable with those of full scan GC-MS, and it allowed the identification and quantitation of the cyclopropane fatty acids in different foods.

In this study we investigated the oligopeptide pattern in fermented cocoa beans and derived products after simulated gastrointestinal digestion. Peptides in digested cocoa samples were identified based on the mass fragmentation and on the software analysis of vicilin and 21 KDa cocoa seed protein sequences, the most abundant cocoa proteins. Quantification was carried out by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) using an internal standard. Sixty five peptides were identified in the digested samples, including three pyroglutamyl derivatives. The in vitro angiotensin-converting enzyme (ACE)-inhibitory activity of cocoa digests were tested, demonstrating a high inhibition activity, especially for digestates of cocoa beans. The peptides identified were screened for their potential ACE inhibitory activity through an in silico approach, and about 20 di-, three- and tetra-peptides actually present in our samples were predicted as active. Two of the potentially active peptides were chemically synthesized and then assessed for their inhibitory activity by using the ACE in vitro assay. These peptides demonstrated an ACE inhibitory activity, however, that was too weak to explain alone the high activity of cocoa digestates, suggesting a synergic effect of all cocoa peptides. As a whole, results showed that an average chocolate portion (30 g) ensures an amount of peptides after digestion that, assuming complete absorption, could reach almost a complete inhibition of ACE.

(1) Background. The aim of this work is to combine non-invasive imaging with chemical characterization analyses to study original and restoration materials of a late 16th-century painting on a canvas representing the “Coronation of the Virgin with the Saints Ambrose and Jerome”, preserved in the Diocesan archive of Orte, a town in the district of Viterbo (Italy). The diagnostic campaign was addressed to support the restoration activities and the choice of the most suitable cleaning operations. (2) Methods. Both traditional analytical techniques and innovative multispectral imaging were applied to solve the diagnostic issues and best address the restoration of the painting. Specifically, hypercolorimetric multispectral imaging (HMI), X-ray fluorescence spectroscopy (XRF), Fourier transform infrared spectroscopy (FT-IR), optical microscopy, and gas chromatography coupled with mass spectrometry (GC-MS) were combined to obtain information on the general conservation state of the artwork and the characterization of pigments, organic binders, and superimposed materials, these last being particularly important to identify ancient and not-documented restoration intervention, enabling the correct choice of the most suitable and effective cleaning intervention. (3) Results. Multispectral data allowed us to differentiate and map original materials through infrared and ultraviolet false color images and spectral reflectance-based similarity maps, suggesting pigment attribution and focusing point analysis for characterization. This approach was particularly successful to identify and locate the presence of unaltered smalt blue in the first painting coat, which had been covered with other pigments, and to suggest the use of organic dye in mixtures with cinnabar and ochres. Spectroscopic and chromatographic techniques enabled us to identify the painting palette and confirm the use of oil-based binder for the pigments and characterize the altered top layers, made with a natural resin and an animal glue. (4) Conclusions. The characterization of the artwork’s materials was essential to select the most suitable methods and materials for the bio-cleaning, based on bacteria, experimented with during the restoration activities.