Carotenoids were analyzed as previously described and each run lasted 21.5 min (27 (link)). Briefly, dried extracts were redissolved in 1 mL of MtBE–MeOH (1:1), filtered with a 0.22 μm nylon filter (CellTreat, Shirley, MA), and 20 μL was injected into a Waters Alliance 2695 HPLC (Waters Corp., Milford, MA) fitted with a 996 DAD. Carotenoids were separated on a 4.6 × 250 mm, 3 μm particle size, C30 column (YMC Inc., Wilmington, NC) maintained at 35°C. A gradient using solvent A: 60% MeOH, 35% MtBE, 3% water, and 2% (w/v) aqueous ammonium acetate and B: 78% MtBE, 20% MeOH, and 2% (w/v) aqueous ammonium acetate at a flow of 1.3 mL/min was used as follows: 100% A to 64.4% A over 9 min, 64.4% A to 0% A over 5.5 min, a hold at 0% A for an additional 3.5 min, and a switch to 100% A for the remaining 3.5 min to recondition the column. Quantification was achieved using a six-point external calibration curve of lycopene and β-carotene. Adjusted slopes were calculated for other carotenoids based on ratios of their molar extinction coefficient to lycopene, as done previously (35 (link)).