Table 2.

Methodology of biological properties of Sceletium sp.

Aim of the Study	Methodology Used			Ref.
	Plant Preparation	Biological Techniques	Phytochemical Screening
To determine psychological effects of Sceletium tortuosum in an in vivo model	S. tortuosum parts were ground into powder form.	In vivo model, using Male Wistar rats in a double placebo study. Oral administration of extract for 17 days and stress was induced. Behavior was monitored.	N/A	[10]
To investigate anti-HIV and free radical scavenging activity of crude extracts of S. tortuosum.	S. tortuosum parts were ground into powder form. Crude extracts of S. tortuosum prepared using ethanol and ethyl acetate.	Anti-HIV assays FRET-based Sensolyte HIV-1 kit used. Fluorogenic substrate mixed with extracts in a 96 well plate. Fluorescence intensity measured. DPPH assay Extract mixed in ethanol and the absorbance measured.	Total phenolic content Total flavonoids content Total tannin content	[36]
To determine the acute effects of Sceletium tortuosum (Zembrin) in human amygdala	Sceletium (Zembrin)	Sceletium (Zembrin) administrated to humans in a placebo double-blind, cross-over study.	N/A	[48]
To screen for anxiolytic activity of Sceletium tortuosum, using an in vivo zebrafish model	S. tortuosum parts ground into powder form. Crude extracts of S. tortuosum prepared in various solvents.	MTC assay used in different concentrations in the zebrafish model study. Larvae placed in a 48-well plate with the control. MTC plates incubated for 18 h at 28.5 °C.	N/A	[60]
To determine the effects of Sceletium tortuosum in the Chick Anxiety-Depression Model	S. tortuosum parts ground into powder form and water used as a solvent.	Preclinical model assay on Male Silver Laced Wyandotte chick administrated S. tortuosum in various concentrations and induced with stress. Distress vocalizations captured the data.	N/A	[68]