Jiang, Yunnan DMPO Cancer Province, China, in September 2018 and identified by the corresponding author Haifeng Tang. The voucher sample (No. 20180903) was deposited inside the Department of Chinese Materia Medica and Natural Medicines, College of Pharmacy, Air Force Healthcare University, Xi’an, China. three.three. Extraction and Isolation The dried rhizomes of Paris mairei (1.0 kg) had been chopped and refluxed with 70 ethanol (ten.0 L) thrice (every 2 h). The ethanol remedy was mixed and condensed with a vacuum rotary evaporator to obtain a syrupy residue (584.0 g). The extraction was suspended in water (3.0 L) and extracted with identical volume of petroleum ether and water saturated n-BuOH three instances, successively. The water saturated within the n-BuOH layer was vacuum evaporated to give a gummy residue (132.0 g). The crude extraction was 2-Bromo-6-nitrophenol manufacturer separated by silica gel column chromatography and eluted by gradient eluent of CH2 Cl2 MeOH-H2 O (100:0:0, 50:1:0, 20:1:0, 8:1:0.1, six:1:0.1, 8:two:0.two, 7:2.five:0.1, and six.5:3.five:0.1) to supply 13 fractions (Fr.13) based on the TLC evaluation. Fr.13 was separated by silica gel column chromatography and eluted by a gradient eluent of CH2 Cl2 -MeOH-H2 O (8:1:0.1, 8:two:0.2, 7:two.5:0.1, and 6:three:0.1) to obtain Fr.13-1 (1.1 g) and Fr.13-2 (830 mg). Fr.13-1 was eluted by MeOH on a Sephadex LH-20 to acquire rid of pigmentum and separated to Fr.13-1-1 (64 mg), Fr.13-1-2 (57 mg), and Fr-13-1-3 (145 mg) on ODS silica gel. Then, Fr.13-1-1 and Fr.13-1-3 had been isolated by semi-preparative HPLC applying MeCN-H2 O (35:65, 40:60) because the mobile phase at a flow rate of eight.0 mL/min to afford compound 1 (9.1 mg, tR = 24.3 min) and 4 (eight.8 mg, tR = 48.six min), respectively. Fr.11 was eluted by MeOH on a Sephadex LH-20 to take away pigmentum to receive Fr.11-1 (four.2 g), Fr.11-2 (five.0 g), and Fr.11-3 (430 mg). Fr.11-2 wasMolecules 2021, 26,13 ofsubjected to ODS silica gel and purified by a semi-preparative HPLC employing MeCN-H2 O (50:50) as the mobile phase at a flow rate of eight.0 mL/min to afford compound 3 (5.7 mg, tR = 44.1 min) and compound 7 (7.6 mg, tR = 40.2 min). Fr.12 was eluted by CH2 Cl2 -MeOH (20:80) on a Sephadex LH-20 to take away pigmentum and subjected to ODS silica gel to receive Fr.12-1 (125 mg) and Fr.12-2 (670 mg). Then, compound two (26.7 mg, tR = 21.0 min) was presented by semi-preparative HPLC using MeCN-H2 O (60:40) as the mobile phase at a flow price of eight.0 mL/min. Fr.9 was purified by MeOH on a Sephadex LH-20 and separated on ODS silica gel to acquire Fr.9-1 (231 mg), Fr.9-2 (102 mg), and Fr.9-3 (193 mg). The 3 collections had been successively purified by semi-preparative HPLC applying MeCN-H2 O (50:50, 40:60, 40:60) because the mobile phase at a flow price of eight.0 mL/min to acquire compounds five (11.5 mg, tR = 35.3 min), eight (five.5 mg, tR = 38.four min), and 9 (four.6 mg, tR = 28.7 min). Fr.10 was eluted by CH2 Cl2 -MeOH (50:50) on a Sephadex LH-20 to take away pigmentum and subjected to ODS silica gel to obtain Fr.10-1 (75 mg) and Fr.10-2 (one hundred mg). Fr.10-1 and Fr.10-2 had been isolated by semi-preparative HPLC using MeCN-H2 O (75:25) as the mobile phase at a flow rate of eight.0 mL/min to afford compounds 10 (7.five mg, tR = 18.3 min) and six (24.4 mg, tR = 21.six min), respectively. The purity of all compounds was assessed by HPLC as getting extra than 95 . 3.four. Compound Characterization Data Pamaiosides A (1): white amorphous solid, []22D – 95.0 (c 0.05, MeOH); IR (KBr) max (cm-1 ): 3420, 2930, 1080, 990, and 840; positive ESI-MS m/z 995.13 [M Na] , unfavorable ESI-MS m/z 971.28 [M – H]- ; good HR-ESI-MS m/z 995.4824.
