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.3. Extraction and Isolation The dried rhizomes of Paris mairei (1.0 kg) have been chopped and refluxed with 70 ethanol (10.0 L) thrice (every two h). The ethanol answer was mixed and condensed with a vacuum rotary evaporator to receive a syrupy residue (584.0 g). The extraction was suspended in water (three.0 L) and extracted with exact same volume of petroleum ether and water saturated n-BuOH 3 times, successively. The water saturated within the n-BuOH layer was vacuum evaporated to provide a gummy residue (132.0 g). The crude extraction was separated by 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, 8:1:0.1, six:1:0.1, eight:two:0.2, 7:2.5:0.1, and six.five:3.5:0.1) to offer 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 (8:1:0.1, eight:2:0.2, 7:two.5:0.1, and six:three:0.1) to acquire 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 utilizing MeCN-H2 O (35:65, 40:60) as the Aztreonam medchemexpress mobile phase at a flow price of 8.0 mL/min to afford compound 1 (9.1 mg, tR = 24.3 min) and four (eight.eight mg, tR = 48.6 min), respectively. Fr.11 was eluted by MeOH on a Sephadex LH-20 to take away 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 ofPolmacoxib site subjected to ODS silica gel and purified by a semi-preparative HPLC employing MeCN-H2 O (50:50) because the mobile phase at a flow rate of eight.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 obtain Fr.12-1 (125 mg) and Fr.12-2 (670 mg). Then, compound two (26.7 mg, tR = 21.0 min) was offered by semi-preparative HPLC using MeCN-H2 O (60:40) because the mobile phase at a flow rate 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 three collections have been successively purified by semi-preparative HPLC making use of MeCN-H2 O (50:50, 40:60, 40:60) because the mobile phase at a flow price of eight.0 mL/min to receive compounds 5 (11.five mg, tR = 35.3 min), eight (five.5 mg, tR = 38.4 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 get Fr.10-1 (75 mg) and Fr.10-2 (100 mg). Fr.10-1 and Fr.10-2 were isolated by semi-preparative HPLC using MeCN-H2 O (75:25) because the mobile phase at a flow rate of 8.0 mL/min to afford compounds 10 (7.5 mg, tR = 18.three min) and six (24.four mg, tR = 21.six min), respectively. The purity of all compounds was assessed by HPLC as getting a lot 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; optimistic ESI-MS m/z 995.13 [M Na] , adverse ESI-MS m/z 971.28 [M – H]- ; constructive HR-ESI-MS m/z 995.4824.
