I was looking for methods for the carotenoids analysis in the urchins' gonads, however, I found that all methods lacked important information such as the initial sample weight, the ratio of sample to solvent, the time for the sample to settle after adding solvents such as n-hexane or Petroleum Ether. Moreover, there is a wide variety of solvents or mixture of solvents used for the same species and each give different results. Also, as mentioned in Chandrika Liyana-Pathirana et al. (2002), after separation of the carotenoid extract in the hexane layer, the reference says to transfer the hexane to 50 ml volumetric flask and make up the volume with hexane, but my extract was already 65 ml, so when I calculate the total carotenoid with the equation, should I use 50 ml or 65 ml?
I have another question, when I have a treatment of triplicate, to measure carotenoid, should I mix samples from all triplicates or should I measure one individual from each replica and then calculate the mean?
My feeding experiment yielded many variation in the color of gonads within the same treatment. So should I discard the very odd color, for example the red, as it occurred in 2 individuals only who appeared diseased and loosing spines, while the remaining samples fluctuated between medium orange roe to dull orange?
I would much appreciate your help with the above specific details in order to be able to run an accurate analysis.
Thank you in advance.
Dear Dr. Elzahrae Elmasry
This article (2012) should help you
Article Extraction and analysis of carotenoids from the New Zealand ...
Garama D, Bremer P, Carne A. Extraction and analysis of carotenoids from the New Zealand sea urchin Evechinus chloroticus gonads. Acta Biochim Pol. 2012;59(1):83-5.
Garama D
Department of Biochemistry, University of Otago, Dunedin, New Zealand. [email protected]
An in case you have many missing questions that need answers, do not hesitate to send a personal message via inbox to any of the corresponding authors or co-authors through RG or even by email.
Best wishes,
Thank you for your kind help. I have this reference, however, it misses the details I am asking for.
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