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Objective To study the sesquiterpenes from the aerial parts of Hyssopus cuspidatus. Methods The compounds were isolated and purified by a combination of various chromatographic techniques including D101 macroporous adsorption resin, silica gel, ODS, Toyopearl HW-40C, Sephadex LH-20, and semi-preparative HPLC. Their structures were elucidated on the basis of physicochemical properties and spectroscopic data. Results Fifteen sesquiterpenes were isolated from the 95% EtOH extracts of H. cuspidatus and their structures were elucidated as 3-eudesmene-1β-11-diol (1), 4-eudesmene-1β,11-diol (2), 1β,11-dihydroxy-5- eudesmene (3), ent-4(15)-eudesmen-1α,11-diol (4), 7α,11-dihydroxy-cadin-10(14)-ene (5), eudesmane-1β,5α,11-triol (6), 5-epi- eudesma-4(15)-ene-1β,6β-diol (7), 4(15)-eudesmene-1β,6α-diol (8), (7R*)-opposit-4(15)-ene-1β,11-diol (9), cryptomeridiol (10), octahydro-4-hydroxy-3α-methyl-7-methylene-α-(1-methylethyl)-1H-indene-1-methanol (11), caryolane-1,9β-diol (12), (3S,5R,6S,7E)- 3,5,6-trihydroxy-7-megastigmen-9-one (13), spathulenol (14), and 4β,10α-aromadendranediol (15), respectively. Conclusion Compounds 1-15 are isolated from the plants of Hyssopus genus for the first time.
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Objective To investigate the optimal process conditions for the separation and purification of extract from Hyssopus cuspidatus Boriss. by polyamide resins. Methods The total flavonoids and rosmarinic acid were used as the indexes. The maximum amount of sample solution, elution volume, concentration of sample solution, adsorption time of resin, loading time of sample solution and the amount of eluting solvent, pH and elution rate in the resin purification process were screened by single factor method. Results The optimal purification parameters were as follows: 10 mg/mL of extract, 12 mL of sample amount, 2 BV of water to remove impurities, 40% ethanol to elute 9 BV; the concentration of rosmarinic acid in sample solution was 86.3 μg/mL, and the total flavonoid concentration was 117.8 μg/mL; the resin adsorption time was 14 h; the pH of sample solution was 6.5; the elution rate was 3.0 BV/h. Conclusion This method is simple and feasible, fit for separating and purifying of extract from Hyssopus cuspidatus Boriss.
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OBJECTIVE:To provide reference for clarifying the pharmacodynamic material basis of anti-inflammation effect of Uygur medicine Hyssopus cuspidatus. METHODS:HPLC was conducted to establish the fingerprint of 8 extracts with different po-larities. Using macrophage RAW264.7 as object,the inhibitory effect of extracts with different polarities on inflammatory factor ni-tric oxide(NO),TNF-α and IL-6 in supernatant of cell culture medium was detected,the differences of anti-inflammatory activity in vitro were compared. Grey correlation analysis method was used to analyze the spectrum-effect relationship of peak area of common peaks and anti-inflammation activity. RESULTS:The fingerprint of 8 extracts with different polarities showed obvious differences. Anti-inflammation in vitro results suggested that 30% ethanol extract had the strongest inhibitory effect on inflammatory cytokines in vitro. There were 14 common peaks in the established HPLC fingerprints,the 5 common peaks that was closely related to the in-hibitory effect of inflammatory factors in vitro were peak 8,6,5,10,13,respectively;peak 8 was rosmarinic acid. CONCLUSIONS:The established fingerprint and its spectrum-effect relationship with anti-inflammation activity in vitro can provide certain reference for its pharmacodynanic material basis study.
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OBJECTIVE:To investigate the effects of different elutions of Hyssopus cuspidatus on smooth muscle contraction of isolated tracheal in Guinea pigs. METHODS:Isolated tracheal rings were prepared and soaked in Krebs-Henseleit,using acetyl-choline(ACh,1×10-7 g/mL)or histamine(His,1×10-6 g/mL)to induce contraction of tracheal rings,then the effects of H. cuspi-datus water elution and 30%,50%,60%,70%,95%ethanol elutions with mass concentrations of 0.08,0.16,0.32,0.64,1.28, 2.56 mg/mL on contraction of tracheal rings were respectively investigated. Contraction curves were recorded and antispasmodic rates were calculated. Tests were treated with saline as blank control and aminophylline (0.08 mg/mL) as positive control. RE-SULTS:Compared with blank control,0.16-2.56 mg/mL 30%,50% ethanol elution,0.32-2.56 mg/mL 60% ethanol elution and 0.64-2.56 mg/mL 70%,95% ethanol elution can obviously inhibit ACh-induced contraction of tracheal rings,antispasmodic rates were obviously increased(P<0.05 or P<0.01);0.32-2.56 mg/mL water elution and 30%,50% ethanol elution,0.16-2.56 mg/mL 60%,70% ethanol elution and 1.28-2.56 mg/mL 95% ethanol elution can obviously inhibit His-induced contraction of tracheal rings,antispasmodic rates were obviously increased (P<0.05 or P<0.01). The effects of 2.56 mg/mL 60% ethanol elution ap-proach to aminophylline. CONCLUSIONS:Different elutions of H. cuspidatus has certain antagonistic effect on the ACh-induced or His-induced smooth muscle contraction of isolated tracheal in Guinea pigs;60% ethanol elution shows the strongest effect, which has similar effects with aminophylline at high mass concentration.