Combination of Analytical and Statistical Methods in Order to Optimize Antibacterial Activity of Clary Sage Supercritical Fluid Extracts

Molecules. 2021 Oct 26;26(21):6449. doi: 10.3390/molecules26216449.


The extraction of clary sage (Salvia sclarea L.) using supercritical carbon dioxide (SC-CO2) was systematically studied by using thin layer chromatography-direct bioautography (TLC-DB) and response surface methodology (RSM). The three parameters temperature, pressure, and cosolvent ratio were optimized for the maximum antibacterial activity of clary sage extracts against Pseudomonas aeruginosa (P. aeruginosa) and methicillin-resistant Staphylococcus aureus (MRSA). The highest inhibition zone was 7.51 mm for P. aeruginosa and 7.57 mm for MRSA. According to RSM analysis, the predicted optimum extraction parameters are 18.6 MPa pressure, 40 °C temperature, and 2% ethanol (EtOH) ratio. The combination of this analytical and statistical method allows saving time, money, and instrument runtime in the optimization of essential oil composition, which is tailored to a specific task and could be useful on any kind of herbs in a wide range of use from perfume manufacturing to the food industry.

Keywords: clary sage; essential oils; response surface method; supercritical fluid extraction; thin layer chromatography-direct bioautography.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification
  • Anti-Bacterial Agents / pharmacology*
  • Carbon Dioxide / chemistry*
  • Chromatography, Supercritical Fluid*
  • Methicillin-Resistant Staphylococcus aureus / drug effects*
  • Microbial Sensitivity Tests
  • Pseudomonas aeruginosa / drug effects*
  • Salvia / chemistry*


  • Anti-Bacterial Agents
  • Carbon Dioxide