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Clin Chim Acta. 2009 Mar;401(1-2):114-8. doi: 10.1016/j.cca.2008.11.022. Epub 2008 Nov 30.

Rapid quantification of steroid patterns in human serum by on-line solid phase extraction combined with liquid chromatography-triple quadrupole linear ion trap mass spectrometry.

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Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Leipzig, Germany.



The determination of steroids is important for the diagnosis and monitoring of endocrine diseases and infertility workup. We developed a rapid and reliable mass spectrometric method for the simultaneous quantification of steroid patterns in human serum.


An on-line solid phase extraction (SPE)-liquid chromatography-triple quadrupole linear ion trap (LC-QTrap) method utilizing atmospheric pressure chemical ionization was developed. Following protein precipitation of 100 microL serum, on-line SPE and chromatographic separation was performed for 13 steroids in 1.8 min. Analytes were confirmed by the characteristic fragment patterns.


The total run time of the method was 4 min. Detection limits ranged between 0.02 microg/L (testosterone) and 9 microg/L (dehydroepiandrosterone sulfate). The method was linear up to 7000 microg/L for dehydroepiandrosterone sulfate, 500 microg/L for cortisol, 125 microg/L for 11-deoxycortisol, and 25 microg/L for aldosterone, 17-hydroxyprogesterone, progesterone, testosterone, androstenedione and beta-estradiol, respectively. Accuracy ranged between 80 and 114%. Between-day variance at three different concentration levels was <15%. Excellent correlations with immunoassays were observed for testosterone, cortisol and beta-estradiol with Pearson's correlation coefficient r=0.967, 0.963, and 0.998, respectively.


The novel on-line SPE-LC-MS/QTrap platform offers a very fast, reliable, and sensitive quantification of steroid patterns and fulfils the quality criteria for routine laboratory application.

[Indexed for MEDLINE]

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