Jiang, Yunnan Province, China, in September 2018 and identified by the corresponding author Haifeng Tang. The voucher sample (No. 20180903) was deposited in the Department of Chinese Materia Medica and All-natural Medicines, School of Pharmacy, Air Force Healthcare University, Xi’an, China. 3.three. Extraction and Isolation The dried rhizomes of Paris mairei (1.0 kg) were chopped and refluxed with 70 ethanol (ten.0 L) thrice (each 2 h). The ethanol resolution was mixed and condensed using a vacuum rotary evaporator to get a syrupy residue (584.0 g). The extraction was suspended in water (3.0 L) and extracted with same volume of petroleum ether and water saturated n-BuOH 3 times, successively. The water saturated in the n-BuOH layer was vacuum evaporated to offer a gummy residue (132.0 g). The crude extraction was separated by Mouse custom synthesis silica gel column chromatography and eluted by gradient eluent of CH2 Cl2 MeOH-H2 O (one hundred:0:0, 50:1:0, 20:1:0, eight:1:0.1, six:1:0.1, 8:two:0.2, 7:2.5:0.1, and 6.five:3.five:0.1) to supply 13 fractions (Fr.13) according to the TLC analysis. 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, 8:2:0.2, 7:two.5:0.1, and 6:3:0.1) to have 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 obtain 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 have been isolated by semi-preparative HPLC applying MeCN-H2 O (35:65, 40:60) as the mobile phase at a flow price of 8.0 mL/min to afford compound 1 (9.1 mg, tR = 24.three min) and 4 (eight.8 mg, tR = 48.6 min), respectively. Fr.11 was eluted by MeOH on a Sephadex LH-20 to remove pigmentum to obtain Fr.11-1 (4.two g), Fr.11-2 (5.0 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 working with MeCN-H2 O (50:50) because the mobile phase at a flow price of 8.0 mL/min to afford compound 3 (5.7 mg, tR = 44.1 min) and compound 7 (7.six mg, tR = 40.two 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 obtain Fr.12-1 (125 mg) and Fr.12-2 (670 mg). Then, compound 2 (26.7 mg, tR = 21.0 min) was supplied by semi-preparative HPLC applying MeCN-H2 O (60:40) as the mobile phase at a flow price of 8.0 mL/min. Fr.9 was purified by MeOH on a Sephadex LH-20 and separated on ODS silica gel to MRTX-1719 supplier receive 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 employing MeCN-H2 O (50:50, 40:60, 40:60) because the mobile phase at a flow rate of eight.0 mL/min to acquire compounds 5 (11.5 mg, tR = 35.three min), 8 (5.5 mg, tR = 38.four min), and 9 (four.6 mg, tR = 28.7 min). Fr.ten was eluted by CH2 Cl2 -MeOH (50:50) on a Sephadex LH-20 to remove 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 making use of MeCN-H2 O (75:25) because the mobile phase at a flow rate of 8.0 mL/min to afford compounds ten (7.five mg, tR = 18.3 min) and six (24.4 mg, tR = 21.6 min), respectively. The purity of all compounds was assessed by HPLC as becoming more than 95 . three.four. Compound Characterization Data Pamaiosides A (1): white amorphous strong, []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]- ; constructive HR-ESI-MS m/z 995.4824.
