Sex hormone and steroid precursor measurement by simple and rapid liquid chromatography-tandem mass spectrometry (Lc-Ms/Ms) method: comparison with current routine immunoassays

Fanelli, Flaminia; Mezzullo, Marco; Fazzini, Alessia; Baccini, Margherita; Gambineri, Alessandra; Vicennati, Valentina; Pelusi, Carla; Pasquali, Renato; Pagotto, Uberto

doi:10.1530/endoabs.41.ep658

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“…The serum steroid profile was assayed by two liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. The first method included cortisone (E), F, 11-deoxycortisol (11 S), 17αOH-progesterone (17αOH-P4), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A), and T [23], while the second included estrone (E1), E2, DHT, and 17αOH-pregnenolone (17αOH-P5) [24]. Four serum samples were analyzed for each of the 21 eligible subjects, and all samples were measured in two consecutive runs.…”

Section: Steroid Measurementsmentioning

confidence: 99%

“…Four serum samples were analyzed for each of the 21 eligible subjects, and all samples were measured in two consecutive runs. Due to analytical specificity requirements, as previously detailed [23,24], data were discarded relating to 17αOH-P5 in five samples from four subjects, to P4 in three samples from one subject of E2 in three samples from three subjects, and to DHT in one sample. In addition, a very small serum volume was available for one sample, resulting in the sensitive measurement of only the T value.…”

Section: Steroid Measurementsmentioning

confidence: 99%

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Impact of Clomiphene Citrate on the Steroid Profile in Dysmetabolic Men with Low Testosterone Levels

et al. 2021

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Clomiphene citrate (CC) in male hypogonadism increases testosterone (T) and estrogen levels by stimulating pituitary gonadotropin release. Our group confirmed these hormonal changes in a randomized, cross-over, double-blind trial of CC versus placebo in addition to metformin, conducted in 21 obese dysmetabolic men with low T levels. However, we hypothesize that based on its mechanism of action, CC may directly or indirectly affect adrenal steroidogenesis. The aim of this sub-study was to better understand the changes in steroid levels and metabolism induced by CC treatment. We assessed 17α-hydroxypregnelone (17αOH-P5), dehydroepiandrosterone (DHEA), progesterone (P4), 17α-hydroxyprogesterone (17αOH-P4), androstenedione (A), T, dihydrotestosterone (DHT), estrone (E1), 17β-estradiol (E2), 11-deoxycortisol (11 S), cortisol (F), and cortisone (E) by LC-MS/MS, and corticosteroid binding globulin (CBG) by ELISA, before and after each treatment. In addition, free-F and steroid product/precursor ratios were calculated. We observed a significant change in serum levels induced by CC compared with placebo for 17αOH-P4, DHT, T, E2, E1, F, E, and CBG, but not free-F. In addition, compared to placebo, CC induced higher 17αOH-P4/P4, E2/E1, 17αOH-P4/17αOH-P5, A/17αOH-P4, T/A, E1/A, F/11 S, and F/E ratios. Therefore, besides the CC stimulating effect on testis steroidogenesis, our study showed increased F, E, but not free-F, levels, indicating changes in steroid metabolism rather than adrenal secretion stimulation. The steroid profiling also revealed the CC stimulation of the Δ5 rather than the Δ4 pathway, thus indicating considerable testicular involvement in the increased androgen secretion.

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Section: Steroid Measurementsmentioning

confidence: 99%

Section: Steroid Measurementsmentioning

confidence: 99%