Steroid hormone profiling in human breast adipose tissue using semi-automated purification and highly sensitive determination of estrogens by GC-APCI-MS/MS

Kristin Hennig, Jean Philippe Antignac, Emmanuelle Bichon, Marie Line Morvan, Isabelle Miran, Suzette Delaloge, Jean Feunteun, Bruno Le Bizec

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26 Citations (Scopus)

Abstract

Body mass index is a known breast cancer risk factor due to, among other mechanisms, adipose-derived hormones. We developed a method for steroid hormone profiling in adipose tissue to evaluate healthy tissue around the tumor and define new biomarkers for cancer development. A semi-automated sample preparation method based on gel permeation chromatography and subsequent derivatization with trimethylsilyl (TMS) is presented. Progestagens and androgens were determined by GC-EI-MS/MS (LOQ 0.5 to 10 ng/g lipids). For estrogen measurement, a highly sensitive GC-APCI-MS/MS method was developed to reach the required lower limits of detection (0.05 to 0.1 ng/g lipids in matrix, 100–200 fg on column for pure standards). The combination of the two methods allows the screening of 27 androgens and progestagens and 4 estrogens from a single sample. Good accuracies and repeatabilities were achieved for each compound class at their respective limit of detection. The method was applied to determine steroid hormone profiles in adipose tissue of 51 patients, collected both at proximity and distant to the tumor. Out of the 31 tested steroid hormones, 14 compounds were detected in human samples. Pregnenolone, 17-hydroxypregnenolone, dehydroepiandrosterone (DHEA), and androstendione accounted together for 80% of the observed steroid hormone profiles, whereas the estrogens accounted for only 1%. These profiles did not differ based on sampling location, except for ß-estradiol; steroid hormone conversions from androgens to estrogens that potentially take place in adipose or tumoral tissue might not be detectable due a factor 100 difference in concentration of for example DHEA and ß-estradiol. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)259-275
Number of pages17
JournalAnalytical and Bioanalytical Chemistry
Volume410
Issue number1
DOIs
Publication statusPublished - 1 Jan 2018
Externally publishedYes

Keywords

  • Adipose tissue
  • Atmospheric pressure chemical ionization
  • Breast cancer
  • GC-APCI-MS/MS
  • Gas chromatography
  • Mass spectrometry
  • Steroid hormones

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