*2.4. Analysis of Carotenoids by HPLC*

Frozen pericarp powder (200 mg) was mixed with 100 μL of 30% NaCl (w:v) solution and 200 μL of dichloromethane to extract carotenoids. Hexane:ether (1:1; 500 μL) was added to the mixture and centrifuged at 13,000× *g* at 4 ◦C for 5 min. This protocol was repeated thrice, and the organic phases were pooled [19]. The upper phase was dried under nitrogen gas and dissolved in ethyl acetate. Samples were analyzed by HPLC (Analytical HPLC, 1260 Infinity II LC System; Agilent Technologies, Santa Clara, CA, USA), coupled to a diode array detector (DAD), and equipped with a YMC Carotenoid HPLC C30 (5 μm × 250 mm × 4.6 mm) column [20]. Lycopene, β-carotene, and lutein from Sigma-Aldrich were used as external standards.
