Ea Gulf of Naples, Goralatide Autophagy Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Antarctica Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Off the coast of Crotone, Ionian Sea Gulf of Naples, Western Mediterranean Sea Antarctica Gulf of Naples, Western Mediterranean Sea Antarctica Antarctica Off the coast of Crotone, Ionian Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Off the coast of Crotone, Ionian Sea Western Mediterranean Sea Off the coast of Crotone, Ionian Sea Off the coast of Crotone, Ionian Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Gulf of Naples, Western Mediterranean Sea Antarctica Off the coast of Crotone, Ionian Sea Antarctica Antarctica Antarctica Antarctica Antarctica Antarctica Gulf of Naples, Western Mediterranean Sea Antarctica Antarctica Antarctica Antarctica Antarctica Antarctica Antarctica Gulf of Naples, Western Mediterranean SeaMar. Drugs 2021, 19,five of2.two. Library of Marine Organic Goods Extracts Raw extracts were obtained by an exhaustive extraction of every marine organism with MeOH. Following the evaporation from the solvent by rotavapor, the samples were transferred using a mixture of MeOH/Dichloromethane (DCM) 2:1 (v/v) to partially desalt the extracts by salt precipitation. Soon after the removal of the solvent under nitrogen, the dry extracts had been stored under argon at -80 C until use. In agreement with [8], SPE fractionation was based on the usage of the hydrophobic solid phase of polystyrene ivinylbenzene. Every single marine extract (50 mg for benthic species and 25 mg for microalgae) was loaded as a water suspension, and fractionation was accomplished by a stepwise solvent gradient elution. Briefly, the very first mobile phase used pure water (fraction A) for an effective desalting. Organic compounds had been then eluted in the four subsequent fractions with MeOH:H2 O 1:1 (v/v), Acetonitrile (ACN):H2 O 7:3 (v/v), ACN, and DCM:MeOH 9:1 (v/v) (fractions B). This procedure gave a reproducible and accurate fractionation of the organic chemical classes, from polar to apolar items. The percentage of every single fraction inside the entire extract for phylum is reported in Table three. The distribution on the metabolites inside the enriched fractions for every SPE was monitored by thin layer chromatography (TLC) and 1 H NMR. General, the pre-fractionation platform generated an MNP library composed of 228 enriched fractions and 57 raw extracts to be tested.Table 3. Percentage contribution of each SPE fraction Cinaciguat supplier towards the total loaded extract deemed for the taxonomic groups represented inside the library. Recovery of total organic compounds (OCs) was calculated because the sum with the SPE fractions B . Phylum Bacillariophyta Bryozoa Chlorophyta Chordata (Tunicata) Cnidaria Haptophyta Miozoa Mollusca Ochrophyta Porifera Rhodophyta Frac. A 39.eight 9.three 71.3 8.2 52.six 18.eight 72.3 9.7 44.1 12.9 49.4 44.7 8.1 63.7 7.two 56.1 ten.six 62.0 17.four 71.0 Frac. B 11.7 6.4 3.two two.0 five.0 six.1 two.6 0.8 3.six 1.0 8.4 3.9 two.three 3.9 0.9 1.eight 2.4 four.7 two.1 1.1 Frac. C 11.4 three three.6 1.two eight.0 six.7 three.3 1.7 4.1 1.four 0.9 6.8 four.1 3.9 .four 4.1 3.six three.7 two.4 two.9 Frac. D ten.4 four.2 2.1 0.7 5.8 4.three 3.1 1.8 5.eight 1.5 16.9 eight.4 four.5 2.9 1.3 4.two four.7 three.eight two.7 0.7 Frac. E 15.two ten.four 5.three 2.0 six.six five.0 three.9 1.8 7.0 three.6 7.8 9.0 five.1 five.8 two.four 6.2 six.three five.8 three.0 1.2 To.