May 26, 2020 · After vortexing, the sample was placed in an ultrasonic bath for 30 minutes. Once removed from the bath, the sample was centrifuged for 5 minutes at 5,000 rpm. Once removed from the centrifuge 50 uL of the ethanol extract is diluted with 950 uL of methanol in a standard HPLC vial. The sample is then injected on the HPLC.