And MeOH 3.81 (3H, s, 7OCH3); 13CNMR (CD E1 r. E7). Fr. E5 was subjected to preparative reverse-phase (9:1:1) to provide seven fractions (Fr. 3OD, 100 MHz): 180.three (C4), 167.5 (C7), 163.0 (C5), 158.five (C9), 157.six (C2), 149.four (C4), 148.four (C3), 140.5 20 mm, S-5 , 12 mm; flow rate, ten.0 mL/min; 30 LC (YMC Actus Triart C18 column; 250 (C3), 126.six (C1), 122.1 (C6), 117.9 (C5), 117.four (C2), 107.0 (C10), 103.5 (C1), 99.2 (C6), 93.three (C8), 78.six (C5), 77.7 (C3), 75.0 (C2), 71.5 (C4), 62.3 (C acetonitrile in H2 O for 60 min; UV ADC Linker Chemical custom synthesis detection at 254 nm) to afford compounds 1 (167 mg) (tR = 45.0 min) six), 60.eight (3OCH3), 56.7 (7OCH3): Supporting facts [20,47]. (Figures 1A and 7).Figure 7. Separation procedure of methanol extract from Nymphoides indica. Figure 7. Separation procedure of methanol extract from Nymphoides indica.Quercetin 3,7-dimethyl ether 4 -glucoside: Yellow power, C23 H24 O12 (mol. wt. 492); HR-ESI-MS (positive ion mode) m/z 493.1346 [M + H]+ , 1 H-NMR (CD3 OD, 400 MHz): 7.63 (1H, d, J = 2.0 Hz, H-2), 7.59 (1H, dd, J = 2.0, eight.4 Hz, H-6), 7.31 (1H, d, J = 8.4 Hz, H-5), 6.57 (1H, d, J = 2.0 Hz, H-8), 6.31 (1H, d, J = two.0 Hz, H-6), 4.94 (1H, d, J = 7.two Hz, H-1″), three.4.8 (6H, m, protons of sugar celebration), 3.88 (3H, s, 3-OCH3), 3.81 (3H, s, 7-OCH3); 13 C-NMR (CD3 OD, one hundred MHz): 180.three (C-4), 167.5 (C-7), 163.0 (C-5), 158.5 (C-9),Molecules 2018, 23,9 of157.6 (C-2), 149.4 (C-4), 148.four (C-3), 140.five (C-3), 126.6 (C-1), 122.1 (C-6), 117.9 (C-5), 117.four (C-2), 107.0 (C-10), 103.five (C-1″), 99.two (C-6), 93.3 (C-8), 78.6 (C-5″), 77.7 (C-3″), 75.0 (C-2″), 71.five (C-4″), 62.3 (C-6″), 60.8 (3-OCH3), 56.7 (7-OCH3): Supporting info [20,47]. three.five. Cell Dopamine Transporter custom synthesis Culture and UVB Irradiation Immortalized human keratinocytes (HaCaT) have been bought in the American Form Culture collection (Manassas, VA, USA). The cells have been cultured in high-glucose DMEM containing 10 FBS, 1 streptomycin-penicillin at 37 C inside a 5 CO2 humidified atmosphere. The cells have been exposed to UVB radiation applying an UV irradiation system (BIO-LINK Crosslinker, WA, Wembley, Australia) delivering the 28020 nm wavelength variety, with maximum emission at 312 nm. Seeded cells were rinsed with PBS after which exposed to 20 mJ/cm2 of UVB. 3.six. Cell Migration HaCaT cells have been incubated, at 5 105 cells/mL for 24 h, in a cell culture incubator. Next, the cell monolayers have been scratched having a 200- yellow tip and washed when with phosphate-buffered saline (PBS). Subsequent, cell monolayers have been treated with distinct concentrations of QDG (1, 5, and 10 /mL) and cultured within a CO2 incubator for 24 h. Cell motility was assessed 24 h later, employing a photomicroscope, and also the scratched region was measured. Measurements had been taken to establish the distance traveled, within the 24 h period, by measuring the scratched location within the photographed pictures. three.7. Immunoassays for Cytokines and Chemokines HaCaT cells (1 105 cells/300 or five 105 cells/400 for the cytokine or chemokine assay, respectively) had been grown within a 24-well plate and treated with UVB (20 mJ/cm2). Immediately after centrifugation at 412g for ten min, the amounts of TNF-, IL-1, IL-6, IL-8, MDC and TARC inside the culture supernatant were analyzed working with the corresponding enzyme-linked immunosorbent assay (ELISA) kits, in line with the manufacturer’s directions. The absorbance was measured at 450 nm utilizing a microplate reader (Magellan; Tecan Ltd, Salzburg, Austria). three.8. Measurement of Skin Barrier Peptide and Hyaluronic Acid HaCaT cells had been seeded in six-well plates, at a de.