R 210 rotavapor evaporator

The extraction method was carried out according to Mandal et al. [20 ]. Briefly, the leaves of each cultivar were air-dried and crushed. One hundred grams of the resulting powder was macerated in methanol at room temperature for 48 h, with repeated maceration until exhaustion. The extracts were then filtered using Whatman No. 1 filter paper and labeled as follows: LRE (extract from leaves of the cultivar with red bracts), LWE (extract from leaves of the cultivar with white bracts), LOE (extract from leaves of the cultivar with orange bracts), LPE (extract from leaves of the cultivar with pink bracts), and LME (extract from leaves of the cultivar with magenta bracts). The extracts were concentrated via vacuum evaporation using a Buchi^® R-210 Rotavapor^® Evaporator (manufactured in Switzerland) at 45 °C.

To extract coral metabolites, ice-cold methanol was added to aliquots of frozen tissue homogenates collected during the 2014 diel study in a ratio of 4:1 (v/v). After sonicating the coral extracts on ice for 2 min, they were freeze-thawed in liquid N₂ three times, and centrifuged (13,000 g, 15 min, 4 °C). The pellet, which contains insoluble materials and proteins, was discarded, and the supernatant was transferred to another container and evaporated to dryness using a Rotary Evaporator (Buchi R-210 Rotavapor Evaporator). To further remove insoluble debris, the dried extracts were reconstituted in MeOH:H₂O (1:1, v/v), vortexed, and centrifuged (13,000 g, 15 min, 4 °C). The supernatants were stored at −80 °C until LC–MS analyses were performed. Additional A. yongei branches processed in an identical manner were used in assays designed to confirm the identity of the corals’ strombine-like compound.