PuSH - Publikationsserver des Helmholtz Zentrums München: Fast separation and quantification of steroid hormones Δ4- and Δ7-dafachronic acid in <em>Caenorhabditis elegans</em>.

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Witting, M. ; Rudloff, H.-C. ; Thondamal, M.* ; Aguilaniu, H.* ; Schmitt-Kopplin, P.

Fast separation and quantification of steroid hormones Δ4- and Δ7-dafachronic acid in Caenorhabditis elegans.

J. Chromatogr. B 978-979, 118-121 (2014)

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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.

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Separation of isomeric molecular species, e.g. double bond position isomers, is a challenging task for liquid chromatography. The two steroid hormones Δ4- and Δ7-dafachronic acid (DA) represent such an isomeric pair. DAs are 3-ketosteroids found in the nematode Caenorhabditis elegans and generated from cholesterol. Δ4- and Δ7-DA have important biological activities and are produced by two different biological pathways in C. elegans. Here we have described a fast separation method for these two isomers using a 1.3μm core-shell particle in less than 10min together with a simple MeOH extraction. Using this method we were able to independently quantify Δ4- and Δ7-DA in C. elegans independently from each other and limits of detection of about 5ng/ml for each isomer were achieved with a good day-to-day reproducibility. As proof-of-principle the method has been applied to the quantification of DAs in worms fed ad libitum or under bacterial deprivation.

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