Dried extracts were redissolved in 1 mL MtBE–MeOH (1:1), filtered with a 0.22 μm nylon filter (CellTreat) and 5 μL was injected into an 1290 Infinity II UHPLC-DAD (Agilent; Santa Clara, CA). Carotenoids were separated on a C18 Acquity BEH column (Waters Corp.) 2.1 × 150 mm, 1.7 μm particle size, maintained at 55°C. An isocratic flow using 42% solvent A [80% MeOH, 20% water, and 2% (w/v) aqueous ammonium acetate] and 58% solvent B [78% MtBE, 20% MeOH, and 2% (w/v) aqueous ammonium acetate] at a flow rate of 0.45 mL/min was used, and each run lasted 4.2 min. Quantification was achieved by six-point external calibration curves as described above. Carotenoid identities were confirmed by authentic standards, spectral characteristics, and tandem MS using a 6495 triple quadrupole MS (Agilent) with an atmospheric pressure chemical ionization source operated in positive mode. Source parameters and multiple reaction monitoring experiments were adapted from those previously reported (27 (link)) and were as follows: phytoene: 545.5>463.6, 421.6, 395.6, 327.4; phytofluene: 543.5>461.6, 393.6, 325.4; β-carotene: 537.5>455.3, 269.2, 69.0; and lycopene: 537.5>455.3, 269.2, 69.0.