Serum steroid profiling by isotopic dilution-liquid chromatography–mass spectrometry: Comparison with current immunoassays and reference intervals in healthy adults

Fanelli, Flaminia; Belluomo, Ilaria; Lallo, Valentina Diana Di; Cuomo, Gaia; Iasio, Rosaria De; Baccini, Margherita; Casadio, Elena; Casetta, Bruno; Vicennati, Valentina; Gambineri, Alessandra; Grossi, G.; Pasquali, Renato; Pagotto, Uberto

doi:10.1016/j.steroids.2010.11.005

Cited by 174 publications

(173 citation statements)

References 35 publications

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“…Immunoassays tend to overestimate the serum concentration of Adione when compared to LC-MS/MS based assays [6], which was also the case in this study (see Fig. 4B).…”

Section: Sample Preparation Referencessupporting

confidence: 76%

TurboFlow-LC-MS/MS method for quantification of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children

Søeborg¹,

Frederiksen²,

Johannsen³

et al. 2013

EJEA

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“…Immunoassays tend to overestimate the serum concentration of Adione when compared to LC-MS/MS based assays [6], which was also the case in this study (see Fig. 4B).…”

Section: Sample Preparation Referencessupporting

confidence: 76%

TurboFlow-LC-MS/MS method for quantification of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children

Søeborg¹,

Frederiksen²,

Johannsen³

et al. 2013

EJEA

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“…Measurement of DHEAS, a hormone of predominantly adrenal origin in women, is not routinely required, although when a neoplastic source of androgen excess is suspected a very high DHEAS concentration may be predictive of an androgen-secreting adrenal cancer (19). The evaluation of steroid hormone profiles, by using liquid chromatography combined with tandem mass spectrometry, a recent technique displaying good precision, sensitivity, and high accuracy, would greatly improve the assessment of androgen excess in patients with PCOS (20,21,22,23).…”

Section: European Journal Of Endocrinologymentioning

confidence: 99%

European survey of diagnosis and management of the polycystic ovary syndrome: results of the ESE PCOS Special Interest Group's Questionnaire

Conway¹,

Dewailly²,

Diamanti-Kandarakis³

et al. 2014

European Journal of Endocrinology

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Background: There is evidence for differences between endocrinologists and other specialists in their approach to diagnosis and management of the polycystic ovary syndrome (PCOS). Objective: A mailed survey consisting of a simple questionnaire aiming to understand current practice for diagnosis and management of the PCOS by specialists across Europe. Methods: The questionnaire consisted of 23 questions grouped to achieve information on i) the general characteristics of the respondents, ii) patients with PCOS seen by endocrinologists, iii) the main diagnostic criteria, iv) biochemical parameters used in the differential diagnosis of hyperandrogenism, v) long-term concerns, and, finally vi) treatment choices. A total of 357 questionnaires representing 13.3% of the members of European Society of Endocrinology (ESE) were available for final analysis; 93% of the respondents were endocrinologists Results: In relation to the diagnostic criteria, respondents were most likely to select menstrual irregularity as the most frequent criteria used for the diagnosis of PCOS although very high rates were achieved for the use of hirsutism and biochemical hyperandrogenism. It therefore appears that the NIH criteria were followed by the majority of respondents. The most frequent biochemical parameters in the differential diagnosis of hyperandrogenism were total testosterone or free androgen index. Obesity and type 2 diabetes were regarded as the principal long-term concerns for PCOS. The most common treatments for patients with PCOS were metformin (33%), lifestyle modification (25%), and oral contraceptives (22%). More direct treatments of infertility include clomiphene citrate alone or in combination with metformin, prescribed by 9 and 23%, respectively, whereas only 6% used other methods for induction of ovulation.

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“…Briefly, samples were purified by solid-phase extraction and injected into a Serie 200 HPLC (PerkinElmer) configured in a two-dimensional LC consisting of a first purification on a perfusion column and a second separation on a Luna RP-C8 100 × 4.6 mm, 5 μm (Phenomenex, Torrance, CA, USA) (22). Afterward, analytes underwent atmospheric pressure chemical ionization and multiple reaction monitoring detection by an API4000 QTrap mass spectrometer (AB SCIEX, Toronto, ON, Canada) (22). Intra-and inter-assay coefficient of variation (CV) were <10 and <11%, respectively, and accuracy ranged between 83.7 and 106.2% for all the analytes (22).…”

Section: Laboratory Analysesmentioning

confidence: 99%

“…Afterward, analytes underwent atmospheric pressure chemical ionization and multiple reaction monitoring detection by an API4000 QTrap mass spectrometer (AB SCIEX, Toronto, ON, Canada) (22). Intra-and inter-assay coefficient of variation (CV) were <10 and <11%, respectively, and accuracy ranged between 83.7 and 106.2% for all the analytes (22). The sensitivity in serum matrix was 0.156 nmol/L (0.049 ng/mL) for P, 0.236 nmol/L (0.078 ng/mL) for 17OHP, 2.70 nmol/L (0.78 ng/mL) for DHEA, 0.136 nmol/L (0.039 ng/mL for A, and 0.066 nmol/L (0.019 ng/mL) for TS (22).…”

Section: Laboratory Analysesmentioning

confidence: 99%

“…A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been recently validated by us for the simultaneous measurement of nine serum steroids (22). In this study, we applied for the first time this validated technique to analyze basal and hCG-stimulated serum androgens and androgen precursors in patients with KS compared with control subjects.…”

Section: Introductionmentioning

confidence: 99%

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Human chorionic gonadotropin stimulation gives evidence of differences in testicular steroidogenesis in Klinefelter syndrome, as assessed by liquid chromatography–tandem mass spectrometry

Belli

Santi

Leoni

et al. 2016

European Journal of Endocrinology

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Background: Men with Klinefelter syndrome (KS) show hypergonadotropic hypogonadism, but the pathogenesis of hypotestosteronemia remains unclear. Testicular steroidogenesis in KS men was evaluated over three decades ago after human chorionic gonadotropin (hCG) stimulation, but inconclusive results were obtained. Intriguingly, some recent studies show increased intratesticular testosterone concentrations in men with KS. Objective: To analyze serum steroid profile, as a proxy of testicular steroidogenesis, after hCG stimulation in KS compared with control men. Design: A prospective, longitudinal, case-control, clinical trial. Methods: Thirteen KS patients (36 ± 9 years) not receiving testosterone (TS) replacement therapy and 12 eugonadic controls (32 ± 8 years) were enrolled. Serum steroids were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) at baseline and for five consecutive days after intramuscular injection of 5000 IU hCG. Results: Progesterone (P), 17-hydroxyprogesterone (17OHP), TS, and estradiol (E2) showed a significant increase (P < 0.001) after hCG stimulation in both groups. On the contrary, androstenedione (AS) and dehydroepiandrosterone did not increase after hCG stimulation. The 17OHP/P ratio increased in both groups (P < 0.001), the TS/AS ratio (17β-hydroxysteroid dehydrogenase type 3 (17βHSD3) activity) did not increase after hCG in any group, and the E2/TS ratio (aromatase activity) increased significantly in both groups (P = 0.009 in KS and P < 0.001 in controls). Luteinizing hormone decreased after hCG in both groups (P = 0.014 in KS and P < 0.001 in controls), whereas follicle-stimulating hormone decreased only in control men (P < 0.001). Conclusion: This study demonstrates for the first time using LC-MS/MS that Leydig cells of KS men are able to respond to hCG stimulation and that the first steps of steroidogenesis are fully functional. However, the TS production in KS men is impaired, possibly related to reduced hydroxysteroid deydrogenase activity due to an unfavorable intratesticular metabolic state.

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Serum steroid profiling by isotopic dilution-liquid chromatography–mass spectrometry: Comparison with current immunoassays and reference intervals in healthy adults

Abstract: Our ID-LC-MS/MS method proved to be reliable and sensitive in revealing steroid circulating concentrations in adults and in highlighting the limits of routine immunoassays at low concentrations.

Cited by 174 publications

References 35 publications

TurboFlow-LC-MS/MS method for quantification of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children

TurboFlow-LC-MS/MS method for quantification of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children

European survey of diagnosis and management of the polycystic ovary syndrome: results of the ESE PCOS Special Interest Group's Questionnaire

Human chorionic gonadotropin stimulation gives evidence of differences in testicular steroidogenesis in Klinefelter syndrome, as assessed by liquid chromatography–tandem mass spectrometry

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