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Separation of Carotenoids by HPLC.

Reference Number: AA-01755 Created: 08/26/2013 02:45 PM Last Updated: 09/17/2020 03:33 PM

Separation of Carotenoids

                    

                        

Peak: 
1. Astaxanthin
         2. Capsanthin
         3. Lutein
         4. Zeaxanthin
         5. Canthaxanthin
         6. ß-Kryptoxanthin
         7. Echinenone
         8. 15-cis ß-Carotene
         9. 13-cis ß-Carotene
        10. a-Carotene
        11. trans ß-Carotene
        12. 9-cis  ß-Carotene
        13. d-Carotene
        14. Lycopene


Method Conditions
      Column: Cogent  C30, 5µm, 200Å 
      Catalog No.: 71030-25 
      Dimensions: 4.6 x 250 mm
      Solvents: A:81/15/4 methanol / MTBE / DI H2O (v/v
                      B: 6/90/4 methanol / MTBE / DI H2O (v/v)
      Gradient:          

              time (min.)     %B                                  
                                           0      
                                        
90      100 
            Temperature: 20°C
            Flow rate: 1.0 ml/min 
            Detection: UV 450 nm
            Samples: Reference standards of each analyte in a mixture.

     

Discussion: Compounds of the carotenoid family are very  lipophilic and often have subtle differences in structure. Resolution of these compounds can be difficult with a typical C8 or C18 column because  of the similarities. The Cogent C30 stationary phase on the other hand can further differentiate by analyte shape in addition to reversed phase interactions. At lower temperatures, the long  alkyl chains become more  rigid and steric effects become significant, leading to greater selectivity.
Notes: Carotenoids are a broad class of more  than 600 compounds. They can be divided into two types: xanthophylls and carotenes. Xanthophylls contain oxygen in their structure and get their name from the Greek words xanthos (yellow) and phyllon (leaf). Carotenes are unsaturated hydrocarbons which do not contain other elements in their structure.

    

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