Anti-inflammatory effects of supercritical carbon dioxide extract and its isolated carnosic acid from rosmarinus officinalis leaves

Chia Feng Kuo, Jeng De Su, Chun Hung Chiu, Chiung Chi Peng, Chi Huang Chang, Tzu Ying Sung, Shiau Huei Huang, Wen Chin Lee, Charng Cherng Chyau

Research output: Contribution to journalArticlepeer-review

68 Citations (Scopus)


Rosemary (Rosmarinus officinalis) leaves possess a variety of bioactivities. Previous studies have shown that the extract of rosemary leaves from supercritical fluid extraction inhibits the expression of inflammatory mediators with apparent dose-dependent responses. In this study, three different extraction conditions (5000 psi at 40, 60, and 80 °C) of supercritical carbon dioxide (SC-CO2) toward the extraction of antioxidants from rosemary were investigated. Furthermore, simultaneous comparison of the anti-inflammatory properties between rosemary extract prepared from SC-CO 2 under optimal conditions (5,000 psi and 80 °C) and its purified carnosic acid (CA) using lipopolysaccharide (LPS)-treated murine RAW 264.7 macrophage cells was also presented. Results showed that the yield of 3.92% and total phenolics of 213.5 mg/g extract obtained from the most effective extraction conditions showed a high inhibitory effect on lipid peroxidation (IC50 33.4 μg/mL). Both the SC-CO2 extract and CA markedly suppressed the LPS-induced production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), phosphorylated inhibitor-kappaB (P-IκB), and nuclear factor-kappaB (NF-κB)/p65 in a dose-dependent manner. The five major compounds of verbenone, cirsimaritin, salvigenin, carnosol, and CA existing in the SC-CO2 extract were isolated by semipreparative HPLC and identified by HPLC-MS/MS analysis. CA was the most abundant recorded compound and the most important photochemical with an anti-inflammatory effect with an IC50 of 22.5 μM or 7.47 μg/mL presented to the best inhibitory activity on NO production better than that of the 14.50 μg/mL dosage prepared from the SC-CO2 extract. Nevertheless, the effective inhibition of LPS-induced NF-κB signaling in RAW 264.7 cells from the SC-CO2 extract extends the potential application of nutraceutical formulation for the prevention of inflammatory diseases.

Original languageEnglish
Pages (from-to)3674-3685
Number of pages12
JournalJournal of Agricultural and Food Chemistry
Issue number8
Publication statusPublished - Apr 27 2011
Externally publishedYes


  • anti-inflammatory activity
  • carnosic acid
  • murine macrophage cell line RAW 264.7
  • Rosemary
  • supercritical carbon dioxide extraction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Chemistry(all)


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