Materials and Reagents

1. Fresh Capsicum fruit (dissected into pericarp tissue, frozen at -80 °C or lyophilized pericarp tissue)
   
2. N₂ gas
   
3. CHCl₃ (HPLC grade) (Sigma-Aldrich, catalog number: 472476 )
   
4. 2-propanol (HPLC grade) (Sigma-Aldrich, catalog number: 34863 )
   
5. Methanol (HPLC grade) (Sigma-Aldrich, catalog number: 179337 )
   
6. KOH (Sigma-Aldrich, catalog number: P-1767 )
   
7. Methyl-t-butyl ether (MTBE) (HPLC grade) (Sigma-Aldrich, catalog number: 34875 )
   
8. β-carotene (Sigma-Aldrich, catalog number: C4582 )
   
9. Lutein (Sigma-Aldrich, catalog number: 07168 )
   
10. Lycopene (Sigma-Aldrich, catalog number: 75051 )
    
11. Antheraxanthin (CaroteNature, catalog number: 0231 )
    
12. Capsanthin (CaroteNature, catalog number: 0335 )
    
13. Capsorubin (CaroteNature, catalog number: 0413 )
    
14. Zeaxanthin (CaroteNature, catalog number: 0119 )
    
15. Violaxanthin (CaroteNature, catalog number: 0259 )
    
16. β-cryptoxanthin (CaroteNature, catalog number: 0055 )
    
17. Methanolic KOH (add solid KOH crystals to methanol until the solution is saturated)
    

Equipment

1. Farberware soft grips (Food Chopper, model: 83427-93 )
   
2. Polytron generator (Polytron Technologies, model: PT 10-35 )
   
3. Temperature block (with 1.8 ml microfuge tube rack) (Thermolyne Dri-Bath, model: DB-17615 )
   
4. HPLC system equipped with a photodiode array detector and YMC carotenoid column (4.6 x 250 mm) (Waters)
   
5. Bath sonicator (Branson Ultrasonic Cleaner, model: 2510 )
   
6. Centrifuge (capable of 1,000 x g, 5 min at 4 °C)
   
7. Microcentrifuge
   
8. UV/Vis spectrophotometer
   
9. Vortex mixer
   
10. Fume hood
    
11. Nitrogen evaporator (Organomation Associates, model: N-EVAP-112 )
    

Procedure