TY - JOUR
T1 - A quantitative comparison of the commonly used methods for extracting carotenoids from avian plasma
AU - McGraw, Kevin
AU - Tourville, Elizabeth A.
AU - Butler, Michael W.
N1 - Funding Information:
Acknowledgments We thank S. Quinn for assistance in capturing ducklings as well as two anonymous referees for providing helpful comments on the manuscript. Financial support for this study was provided by the School of Life Sciences and College of Liberal Arts and Sciences at Arizona State University. Birds from both species were captured and sampled under university (protocol nos. 05-764R and 07-910R), state (SP797514), and federal (MB088806-0) permits.
PY - 2008/10
Y1 - 2008/10
N2 - Interest in animal carotenoids, especially in birds, has exploded in recent years, and so too have the methods employed to investigate the nature and function of these pigments. Perhaps the most easily and commonly performed procedure in this work has been the determination of carotenoid concentration from avian plasma. Over the past 20 years of research on avian carotenoids, numerous methods have been used to extract carotenoids from bird plasma, all of which have differed in several important parameters (e.g., number and types of solvents used, degree of mixing/centrifugation). However, to date, no study has systematically compared these methods to determine if any of them are more effective than others for recovering any or all types of carotenoids present. We undertook such an investigation on plasma samples from two bird species (house finch, Carpodacus mexicanus, and mallard, Anas platyrhynchos) using five of the most commonly employed methods for extracting carotenoids from avian plasma: (1) acetone-only, (2) methanol-only, (3) ethanol-only, (4) ethanol + hexane, and (5) ethanol + tert butyl methyl ether. We also manipulated the amount of time that extracts were centrifuged, which has varied tremendously in previous studies, to evaluate its importance on carotenoid recovery. We found that all methods equally recovered the polar xanthophylls (lutein and zeaxanthin), but that the methanol-only procedure poorly recovered non-polar carotenoids (less β-carotene in both species and less β-cryptoxanthin in house finches) compared to the other methods. These results suggest that the data accumulated to date on xanthophyll plasma carotenoids in birds should be comparable across studies and species despite the different chemical extraction methods used. However, care should be taken to use relatively strong organic solvents for fully recovering non-polar carotenoids. We also found no effect of centrifugation duration (1 vs. 10 min at 10,000 rpm) on carotenoid recoveries, demonstrating that researchers can save considerable time by centrifuging for a much shorter time period than is typically used.
AB - Interest in animal carotenoids, especially in birds, has exploded in recent years, and so too have the methods employed to investigate the nature and function of these pigments. Perhaps the most easily and commonly performed procedure in this work has been the determination of carotenoid concentration from avian plasma. Over the past 20 years of research on avian carotenoids, numerous methods have been used to extract carotenoids from bird plasma, all of which have differed in several important parameters (e.g., number and types of solvents used, degree of mixing/centrifugation). However, to date, no study has systematically compared these methods to determine if any of them are more effective than others for recovering any or all types of carotenoids present. We undertook such an investigation on plasma samples from two bird species (house finch, Carpodacus mexicanus, and mallard, Anas platyrhynchos) using five of the most commonly employed methods for extracting carotenoids from avian plasma: (1) acetone-only, (2) methanol-only, (3) ethanol-only, (4) ethanol + hexane, and (5) ethanol + tert butyl methyl ether. We also manipulated the amount of time that extracts were centrifuged, which has varied tremendously in previous studies, to evaluate its importance on carotenoid recovery. We found that all methods equally recovered the polar xanthophylls (lutein and zeaxanthin), but that the methanol-only procedure poorly recovered non-polar carotenoids (less β-carotene in both species and less β-cryptoxanthin in house finches) compared to the other methods. These results suggest that the data accumulated to date on xanthophyll plasma carotenoids in birds should be comparable across studies and species despite the different chemical extraction methods used. However, care should be taken to use relatively strong organic solvents for fully recovering non-polar carotenoids. We also found no effect of centrifugation duration (1 vs. 10 min at 10,000 rpm) on carotenoid recoveries, demonstrating that researchers can save considerable time by centrifuging for a much shorter time period than is typically used.
KW - Carotenoid pigments
KW - Ethanol
KW - HPLC
KW - House finch
KW - Lutein
KW - Mallard
KW - Methanol
KW - Zeaxanthin
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U2 - 10.1007/s00265-008-0622-4
DO - 10.1007/s00265-008-0622-4
M3 - Article
AN - SCOPUS:51849120016
VL - 62
SP - 1991
EP - 2002
JO - Behavioral Ecology and Sociobiology
JF - Behavioral Ecology and Sociobiology
SN - 0340-5443
IS - 12
ER -