Profiling Compounds in CareVid™ Tea Powder

The nine compounds (1–9) were determined using HPLC-MS to be present in CareVid^TM as described previously [1 (link)]. Extracts were prepared initially from 20 g each of CareVid^TM Tea powder. The extracts were methylene chloride, ethanol, methanol, water, acidified water (0.1 M HCl), and steeping in hot water (>90 °C). Metabolic profiling of the extracts was performed by UHPLC-UV-HRMS/MS on a Vanquish UHPLC system (Thermo Scientific, Waltham, MA, USA) coupled to a 100 Hz photodiode array detector (PDA) and an Orbitrap Fusion Tribrid (Thermo Scientific) high-resolution tandem mass spectrometer (Thermo Scientific, Waltham, MA, USA). A detailed protocol that was followed is described in Rotich et al., 2021 (Figure 1). For the assay, pure standards of pellitorine (1), oleuropein (2) and ellagic acid (9), urolithin A (10), and urolithin B (11) were bought from Sigma Aldrich, whereas pure standards of magnoflorine (3) earlier isolated from the seed cake of Croton megalocarpus, crotocorylifuran (6) earlier isolated from Croton megalocarpoides, crotepoxide (4) earlier isolated from Croton alienus, ent-kaurane-16β,17-diol (5) earlier isolated from Croton haumanianus, and lupeol (7) and betulin (8) isolated in substantial quantities from CareVid^TM were obtained in the Jodrell Laboratory, Kew, UK.