Jiang, Yunnan Province, China, in September 2018 and identified by the corresponding author Haifeng Tang. The voucher sample (No. 20180903) was deposited within the Division of Chinese Materia Medica and All-natural Medicines, School of Pharmacy, Air Force Medical University, Xi’an, China. three.three. Extraction and Isolation The dried rhizomes of Paris mairei (1.0 kg) have been chopped and refluxed with 70 ethanol (ten.0 L) thrice (every single two h). The ethanol remedy was mixed and condensed having a vacuum rotary evaporator to receive a FAUC 365 Biological Activity 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, 6:1:0.1, 8:two:0.two, 7:2.five:0.1, and six.5:3.five:0.1) to give 13 fractions (Fr.13) depending on the TLC analysis. Fr.13 was separated by silica gel column chromatography and eluted by a gradient eluent of CH2 Cl2 -MeOH-H2 O (eight:1:0.1, 8:2:0.two, 7:2.five:0.1, and six:three:0.1) to get 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 have been isolated by semi-preparative HPLC working with MeCN-H2 O (35:65, 40:60) as the mobile phase at a flow rate of 8.0 mL/min to afford compound 1 (9.1 mg, tR = 24.three min) and 4 (8.8 mg, tR = 48.6 min), respectively. Fr.11 was eluted by MeOH on a Sephadex LH-20 to get rid of pigmentum to acquire Fr.11-1 (4.2 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 rate of 8.0 mL/min to afford compound three (5.7 mg, tR = 44.1 min) and compound 7 (7.6 mg, tR = 40.two min). Fr.12 was eluted by CH2 Cl2 -MeOH (20:80) on a Sephadex LH-20 to eliminate pigmentum and subjected to ODS silica gel to receive Fr.12-1 (125 mg) and Fr.12-2 (670 mg). Then, compound 2 (26.7 mg, tR = 21.0 min) was offered 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 receive Fr.9-1 (231 mg), Fr.9-2 (102 mg), and Fr.9-3 (193 mg). The 3 collections have been successively purified by semi-preparative HPLC working with MeCN-H2 O (50:50, 40:60, 40:60) as the mobile phase at a flow rate of 8.0 mL/min to obtain compounds five (11.five mg, tR = 35.three min), 8 (5.5 mg, tR = 38.4 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 eliminate pigmentum and subjected to ODS silica gel to receive Fr.10-1 (75 mg) and Fr.10-2 (100 mg). Fr.10-1 and Fr.10-2 have been isolated by semi-preparative HPLC working with 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.four mg, tR = 21.6 min), respectively. The purity of all compounds was assessed by HPLC as getting much more than 95 . 3.4. 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]- ; optimistic HR-ESI-MS m/z 995.4824.
