BackgroundSpermacoce princeae(K. Schum) has been used in the treatment of bacterial skin infections in Uganda. Pharmacological studies revealed that extracts ofS. princeaeexhibited antibacterial, antioxidant, and sun protection potential. This study aimed at isolating and identifying pure compounds from the extracts based on comprehensive analytical characterization by multiple analytical techniques.MethodsThe plant samples were extracted by sequential maceration usingn-hexane, ethyl acetate, methanol, and distilled water. The compounds were isolated using a combination of chromatographic techniques and their structures were elucidated by multiple spectroscopic techniques. The antibacterial and antifungal activity determination of the isolated compounds was carried out using an agar well diffusion and potato dextrose assay againstPseudomonas aeruginosa,Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Candida albicans, and Aspergillus flavuswhile the antioxidant activity was screened with the 2,2-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay. The sun protection factor was determined using a Shimadzu Ultra Violet-visible (UV–VIS) double beam spectrophotometer between 290 to 320 nm.ResultsEleven compounds; quercetin (1), kaempferol-3-O-rutinoside (2), rutin (3,12),myo-inositol (4), asperulosidic acid (5), hexadecanoic acid (6), β-sitosterol (7), stigmasterol (8), campesterol (9), ursolic acid (10), and β-sitosterol glucoside (11) were identified in theS. princeaeextracts. Compound2had good antifungal activity againstC. albicans(zone of inhibition, 23.0 ± 0.1 mm). Compound10showed antibacterial and antifungal activity againstS.aureus, P. aeruginosa, C. albicans, and A. flavus. Compound2had a good percentage radical scavenging effect (IC50= 64.81 µg/ml) and a good sun protection factor (SPF = 26.83).ConclusionThis study reports the first-time isolation and identification of compounds1to11fromS. princeae,which contribute to its antimicrobial, antioxidant, and sun protection potential.