An instructor in an applied biotechnology laboratory provides oral instructions regarding the extraction and concentration of secondary metabolites from plant tissues:
"Before introducing the dried botanical sample into the extraction vessel, you must thoroughly homogenize the dried leaves using a sterile mortar and pestle. However, prior to performing any grinding, ensure that the ethanol-water solvent mixture has already been chilled to 4°C. Once the pulverized material is submerged in the chilled solvent and macerated for thirty minutes, centrifuge the suspension to separate the solid biomass from the liquid. Finally, only after the liquid supernatant has been completely passed through a fine membrane filter should you transfer it to the rotary evaporator flask for concentration."
Based on the transitional markers and conditional timing in the audio instructions, arrange the following actions in the exact chronological order in which they must be performed.
- 1Verify that the ethanol-water solvent mixture is chilled to 4°C.
- 2Homogenize the dried botanical leaves using a sterile mortar and pestle.
- 3Centrifuge the suspension to separate the solid biomass from the liquid.
- 4Pass the liquid supernatant completely through a fine membrane filter.