Supplementary Materialsmolecules-25-00103-s001. and in the skin. In-source fragmentation of proanthocyanidins was determined using their standards, suggesting their distribution was mixed ion images of themselves, and fragment ions generated from those had a higher degree of polymerization. B-type procyanidins were predominantly distributed in the vascular bundles than in the skin, whereas B-type propelargonidins were almost equally distributed between the vascular bundles and skin, suggesting that their distribution patterns are different from the type of their flavan-3-ol monomers. Flavan-3-ols, especially B-type procyanidins, may help prevent pathogen infection not only in the skin but also in and around the vascular bundles. of 240 to 1200 obtained by MALDI-MSI. Regarding flavan-3-ol monomers, peaks at 273.1 and 289.1 corresponding to (epi)afzelechin and (epi)catechin [M ? H]? ions were observed. The chirality of C-3 in flavan-3-ols cannot be differentiated by the MS analysis performed in this study; therefore, (epi)afzelechin represented either afzelechin or epiafzelechin. Similarly, (epi)catechin represented either catechin or epicatechin. Regarding proanthocyanidins, we observed peaks corresponding to values of B-type procyanidins, namely (epi)catechin dimer, trimer, and tetramer [M ? H]? ions, at 577.1, 865.2, and 1153.3, respectively (Figure 2B). When one (epi)afzelechin was present in proanthocyanidins, the signals of ARQ-092 (Miransertib) procyanidins were ARQ-092 (Miransertib) shifted by ?16 mass units from the signals of B-type procyanidins, corresponding to the loss of one oxygen atom (Figure 1A). Peaks matching the values of B-type propelargonidin [M ? H]? ions with one (epi)afzelechin unit and one or two (epi)catechin units, i.e., peaks at 561.1 and 849.2, respectively, were observed (Figure 2B). Open in a separate window Figure 2 Matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry imaging (MSI) of strawberry fruit sections. (A) Optical image Rabbit Polyclonal to PEA-15 (phospho-Ser104) of a strawberry fruit section before MALDI-MSI analysis. The dotted white line shows the examined area. (B) Mass range from the strawberry fruits section. The values indicated how the peaks were due to flavan-3-ols possibly. The relative strength from the most powerful 289.1 in the peaks with potential substances of (epi)catechin was thought as 95%. (Epi)catechin represents either catechin or epicatechin. Representative ion pictures at (C) 273.1, (D) 289.1, (E) 561.1, (F) 577.1, (G) 849.2, (H) 865.2, and (We) 1153.3. Size pub = 5 mm. To research the distribution of peaks at 273.1, 289.1, 561.1, 577.1, 849.2, 865.2, and 1153.3, that are potential the different parts of the flavan-3-ols monomer and oligomer [M ? H]? ions in ripe strawberry fruits, we built their ion pictures. As demonstrated in Shape 2CCI, all potential the different parts of flavan-3-ols had been distributed in the calyx primarily, around the vascular bundles, and in your skin. 2.3. Recognition of Flavan-3-ols in Strawberry Fruits Areas MALDI-time of trip (TOF)/TOF evaluation allows accurate and extremely delicate elucidation of flavan-3-ols predicated on their MS/MS spectra [7,8,9]. Next, we performed MALDI-MS/MS evaluation to determine if the noticed peaks at 273.1, 289.1, 561.1, 577.1, 849.2, 865.2, and 1153.3 (Shape 2B) had been due to flavan-3-ol [M ? H]? ions. As demonstrated in Shape 3A, the MS/MS spectra had been acquired in the calyx, around the vascular bundles, and in your skin from the strawberry fruits section as the potential the different parts of flavan-3-ols had been mainly distributed in these cells (Shape 2CCI). The MS/MS spectra of potential peaks for flavan-3-ols [M ? H]? ions from the three cells showed identical fragmentation patterns. Open in a separate window Physique 3 Matrix-assisted laser desorption/ionization (MALDI)-tandem mass spectrometry (MS/MS) analysis of strawberry fruit sections. MALDI-MS/MS spectra were obtained from in the calyx, in and around the vascular bundles, and in the skin of strawberry fruit sections after MALDI-mass spectrometry imaging. (A) Optical image of a strawberry fruit section. The dotted red line shows the analyzed region. Scale bar = 5 mm. Representative MS/MS spectra of precursor [M ? H]? ions at (B) 289.1, (C) 561.1, (D) 577.1, (E) 849.2, (F) 865.2, and (G) 1153.3 obtained from a strawberry fruit section. The fragment ion peaks with values were used for molecular assignments. MS/MS analysis of the peak of the afzelechin standard [M ? H]? ions showed fragment ion peaks at ARQ-092 (Miransertib) 257 (loss of water) and 167 (Supplementary Physique S2A). However, in the MS/MS spectra of precursor ion at 273.1 detected in strawberry sections, the fragment ions at 257 and 167 were hardly observed (Supplementary Determine S2B). Therefore, we could not determine whether the peak at 273.1 detected in strawberry sections was attributable to afzelechin and/or epiafzelechin [M ? H]? ions. In the MS/MS spectrum of precursor ions at 289.1, fragment ions at 245 and 125 were observed (Physique 3B). This fragmentation pattern agreed with the MS/MS spectra of both catechin and epicatechin standards (Supplementary.