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Use of LC–MS–MS as an alternative to currently available immunoassay methods to quantitate corticosterone in egg yolk and albumen

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Abstract
Corticosterone (CORT) is the dominant plasma glucocorticoid in birds. There has been increasing interest in the function of CORT in avian egg yolk and in the potential to use CORT concentrations in eggs to quantify stress and to assess the effect of maternal stress on offspring. The concentration of CORT in egg yolk is most frequently assessed using enzyme or radioimmunoassays, alone or in combination with high-performance liquid chromatography. However, the quantification of CORT is frequently hampered by the presence of high concentrations of other steroid hormones which crossreact with the CORT antibody. As an alternative, we developed a sensitive and specific LC–MS–MS method. The sample-preparation procedure consisted of a protein–lipid precipitation step, followed by defatting and clean-up using a C18 SPE column. Chromatography was performed on an Acquity C18 BEH column (50 mm×2.1 mm i.d., dp: 1.7 μm, run-time: 6 min), using 0.1 % formic acid in both water (A) and acetonitrile (B) as mobile phases. The MS–MS instrument was operated in the positive-electrospray-ionization mode. The method was validated in-house according to European Guidelines (linearity, accuracy and precision, limits of quantification and detection, specificity, stability) and the results fell within the accepted ranges. The method was successfully used for the analysis of CORT in yolk and albumen of eggs collected from eight breeding lesser black-backed gulls at a Flemish coastal colony. CORT concentrations were in the range 42.4–166.3 pg g−1 in albumen and<LOQ (75 pg g−1)–762.5 pg g−1 in yolk.
Keywords
Steroid hormones, TANDEM MASS-SPECTROMETRY, Yolk, Albumen, Eggs, LC–MS–MS, Corticosterone, STEROID-HORMONES, LAYING HENS, VALIDATION, ANDROGENS

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Chicago
De Baere, Siegrid, Tom Rosendahl Larsen, Mathias Devreese, Patrick De Backer, Liesbeth De Neve, Graham Fairhurst, Luc Lens, and Siska Croubels. 2015. “Use of LC–MS–MS as an Alternative to Currently Available Immunoassay Methods to Quantitate Corticosterone in Egg Yolk and Albumen.” Ed. Siska Croubels, Els Daeseleire, Sarah De Saeger, Peter Van Eenoo, and Lynn Vanhaecke. Analytical and Bioanalytical Chemistry 407 (15): 4351–4362.
APA
De Baere, Siegrid, Larsen, T. R., Devreese, M., De Backer, P., De Neve, L., Fairhurst, G., Lens, L., et al. (2015). Use of LC–MS–MS as an alternative to currently available immunoassay methods to quantitate corticosterone in egg yolk and albumen. (S. Croubels, E. Daeseleire, S. De Saeger, P. Van Eenoo, & L. Vanhaecke, Eds.)ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 407(15), 4351–4362.
Vancouver
1.
De Baere S, Larsen TR, Devreese M, De Backer P, De Neve L, Fairhurst G, et al. Use of LC–MS–MS as an alternative to currently available immunoassay methods to quantitate corticosterone in egg yolk and albumen. Croubels S, Daeseleire E, De Saeger S, Van Eenoo P, Vanhaecke L, editors. ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 2015;407(15):4351–62.
MLA
De Baere, Siegrid et al. “Use of LC–MS–MS as an Alternative to Currently Available Immunoassay Methods to Quantitate Corticosterone in Egg Yolk and Albumen.” Ed. Siska Croubels et al. ANALYTICAL AND BIOANALYTICAL CHEMISTRY 407.15 (2015): 4351–4362. Print.
@article{5944597,
  abstract     = {Corticosterone (CORT) is the dominant plasma glucocorticoid in birds. There has been increasing interest in the function of CORT in avian egg yolk and in the potential to use CORT concentrations in eggs to quantify stress and to assess the effect of maternal stress on offspring. The concentration of CORT in egg yolk is most frequently assessed using enzyme or radioimmunoassays, alone or in combination with high-performance liquid chromatography. However, the quantification of CORT is frequently hampered by the presence of high concentrations of other steroid hormones which crossreact with the CORT antibody. As an alternative, we developed a sensitive and specific LC–MS–MS method. The sample-preparation procedure consisted of a protein–lipid precipitation step, followed by defatting and clean-up using a C18 SPE column. Chromatography was performed on an Acquity C18 BEH column (50 mm×2.1 mm i.d., dp: 1.7 μm, run-time: 6 min), using 0.1 % formic acid in both water (A) and acetonitrile (B) as mobile phases. The MS–MS instrument was operated in the positive-electrospray-ionization mode. The method was validated in-house according to European Guidelines (linearity, accuracy and precision, limits of quantification and detection, specificity, stability) and the results fell within the accepted ranges. The method was successfully used for the analysis of CORT in yolk and albumen of eggs collected from eight breeding lesser black-backed gulls at a Flemish coastal colony. CORT concentrations were in the range 42.4–166.3 pg g−1 in albumen and<LOQ (75 pg g−1)–762.5 pg g−1 in yolk.},
  author       = {De Baere, Siegrid and Larsen, Tom Rosendahl and Devreese, Mathias and De Backer, Patrick and De Neve, Liesbeth and Fairhurst, Graham and Lens, Luc and Croubels, Siska},
  editor       = {Croubels, Siska and Daeseleire, Els and De Saeger, Sarah and Van Eenoo, Peter and Vanhaecke, Lynn},
  issn         = {1618-2642},
  journal      = {ANALYTICAL AND BIOANALYTICAL CHEMISTRY},
  keywords     = {Steroid hormones,TANDEM MASS-SPECTROMETRY,Yolk,Albumen,Eggs,LC–MS–MS,Corticosterone,STEROID-HORMONES,LAYING HENS,VALIDATION,ANDROGENS},
  language     = {eng},
  number       = {15},
  pages        = {4351--4362},
  title        = {Use of LC–MS–MS as an alternative to currently available immunoassay methods to quantitate corticosterone in egg yolk and albumen},
  url          = {http://dx.doi.org/10.1007/s00216-014-8269-7},
  volume       = {407},
  year         = {2015},
}

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