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<br>Display Omitted<br>• Response surface methodology was applied to optimize supercritical fluid extraction conditions of alantolactone and isoalantolactone. • The optimized extraction conditions were pressure 20 MPa, extraction temperature 50 °C, carbon dioxide flow rate of 17 × 10−5 kg/s and extraction time of 40 min. • Freezing purification greatly improve the content of the target compounds by 30.7 wt.%.<br><b>Inula racemose</b> Hook.f. (<b>I</b>. <b>racemosa</b>) is a traditional herbal medicine with strong anti-fungal and anti-inflammation activity while alantolactone (AL) and isoalantolactone (IAL) are the major active compounds of this plant. The aim of this paper was obtaining AL and IAL from the radix of <b>I</b>. <b>racemosa</b> by supercritical fluid extraction (SFE) which was optimized by response surface methodology (RSM). Our results showed that, the optimal conditions for maximum extraction efficiency were as follows: pressure 20 MPa, extraction temperature 50 °C, carbon dioxide (CO2) flow rate of 17.0 × 10−5 kg/s and extraction time of 40 min. Subsequently, an efficient freezing method was developed for purification of AL and IAL in the crude extract obtained from SFE. After purification by freezing, the total content of the target compounds was greatly improved by 30.7%. The results demonstrate that SFE coupled with freezing would be a powerful technique for extraction and purification of AL and IAL from the radix of <b>I</b>. <b>racemosa</b>.

Five phenylethanoid glycosides (PhGs), forsythoside B, verbascoside, alyssonoside, isoverbascoside, and leucosceptoside B, were isolated and purified from Lamiophlomis rotata (Benth.) Kudo by high-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column separation. In the present study, the two-phase solvent system composed of ethyl acetate/n-butanol/water (13:3:10, v/v/v) was used for HSCCC separation. A total of 27 mg of forsythoside B, 41 mg of verbascoside, 29 mg of alyssonoside, 23 mg of isoverbascoside, and 13 mg of leucosceptoside B with purities of 97.7, 99.2, 99.5, 99.3, and 97.3%, respectively, were obtained in a one-step separation within 4 h from 150 mg of crude extract. The recoveries of the five PhGs after MR-HSCCC separation were 74.5, 76.5, 72.5, 76.4, and 77.0%, respectively. The chemical structures of all five compounds were identified by (1) H and (13) C NMR spectroscopy.

Four iridoid glucosides, shanzhiside methyl ester, phloyoside II, chlorotuberside, and penstemonoside, were isolated and purified from an herbal medicinal plant for the first time by high-speed counter-current chromatography (HSCCC) using a two-phase solvent system composed of ethyl acetate-n-butanol-water (5:14:12, v/v/v). A total of 37mg of shanzhiside methyl ester, 29mg of phloyoside II, 27mg of chlorotuberside, and 21mg of penstemonoside with the purity of 99.2%, 98.5%, 97.3%, and 99.3%, respectively, were obtained in one-step separation within 4h from 150mg of crude extract. To the best of our knowledge, this is the first report of separation and purification of iridoid glucosides from natural sources by HSCCC. The chemical structures of all the four compounds were identified by ESI-MS, (1)H NMR, and (13)C NMR.