Fully automated method for quantitative determination of steroids in serum: An approach to evaluate steroidogenesis

Luque-Córdoba, D.; Priego‐Capote, Feliciano

doi:10.1016/j.talanta.2020.121923

Cited by 10 publications

(7 citation statements)

References 56 publications

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“…On-line SPE has been published frequently for steroids in combination with tandem mass spectrometry [19] , [20] , [21] . This shows the increasing importance of this technique in the field of laboratory medicine where it is already used for endocrinological diagnostics [22] , [23] , [24] .…”

Section: Discussionmentioning

confidence: 99%

Semi-automated serum steroid profiling with tandem mass spectrometry

Rakete¹,

Schubert²,

Vogeser³

2023

Journal of Mass Spectrometry and Advances in the Clinical Lab

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Section: Discussionmentioning

confidence: 99%

Semi-automated serum steroid profiling with tandem mass spectrometry

Rakete¹,

Schubert²,

Vogeser³

2023

Journal of Mass Spectrometry and Advances in the Clinical Lab

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“…The sensitivity of the assay was 5 pg/tube for LH and 20 pg/tube for FSH. In addition, sex steroid levels were measured in serum samples using a previously validated, sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, adapted from protocols described in detail elsewhere (Luque-Cordoba and Priego-Capote 2021). Determinations of estradiol (E 2 ), testosterone (T), dihydro-testosterone (DHT) and progesterone (P) were conducted in control and TaKKO mice of both sexes, at 4-weeks and 2-months of age.…”

Section: Methodsmentioning

confidence: 99%

Dissecting the KNDy hypothesis: KNDy neuron-derived kisspeptins are dispensable for puberty but essential for preserved female fertility and gonadotropin pulsatility

Velasco

Franssen

Daza-Dueñas

et al. 2022

Preprint

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Kiss1 neurons in the hypothalamic arcuate-nucleus (ARC) play key roles in the control of GnRH pulsatility and fertility. A fraction of ARC Kiss1 neurons, termed KNDy, co-express neurokinin B (NKB; encoded by Tac2). Yet, NKB- and Kiss1-only neurons are also found in the ARC, while a second major Kiss1-neuronal population is present in the rostral hypothalamus. The specific contribution of different Kiss1 neuron sub-sets to reproductive control remains unfolded. To tease apart the physiological roles of KNDy-born kisspeptins, conditional ablation of Kiss1 in Tac2-expressing cells was implemented in vivo. Mice with Tac2 cell-specific Kiss1 KO (TaKKO) displayed reduced ARC kisspeptin content and Kiss1 expression, with greater suppression in females, which was detectable at infantile-pubertal age. In contrast, Tac2/NKB levels were fully preserved. Despite the drop of ARC Kiss1/kisspeptin, pubertal timing was normal in TaKKO mice of both sexes. However, young-adult TaKKO females displayed disturbed LH pulsatility and sex steroid levels, with suppressed basal LH and pre-ovulatory LH surges, early-onset subfertility and premature ovarian insufficiency. Conversely, testicular histology and fertility were grossly conserved in TaKKO males. Ablation of Kiss1 in Tac2-cells led also to sex-dependent alterations in body composition, glucose homeostasis and locomotor activity. Our data document that KNDy-born kisspeptins are dispensable/compensable for puberty in both sexes, but required for maintenance of female gonadotropin pulsatility and fertility, as well as adult metabolic homeostasis.Significance StatementNeurons in the hypothalamic arcuate nucleus (ARC) co-expressing kisspeptins and NKB, named KNDy, have been recently suggested to play a key role in pulsatile secretion of gonadotropins, and hence reproduction. However, the relative contribution of this Kiss1 neuronal-subset, vs. ARC Kiss1-only and NKB-only neurons, as well as other Kiss1 neuronal populations, has not been assessed in physiological settings. We report here findings in a novel mouse-model with elimination of KNDy-born kisspeptins, without altering other kisspeptin compartments. Our data highlights the heterogeneity of ARC Kiss1 populations and document that, while dispensable/compensable for puberty, KNDy-born kisspeptins are required for proper gonadotropin pulsatility and fertility, specifically in females. Characterization of this functional diversity is especially relevant, considering the potential of kisspeptin-based therapies for management of human reproductive disorders.Disclosure StatementThe authors have nothing to disclose in relation to the contents of this work.

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“…More importantly, estrogens can be only monitored under the negative ion mode during direct analysis. To achieve adequate sensitivity, ammonium fluoride should be introduced into the mobile phase in many cases. − However, even under neutral conditions, fluoride anions can degrade silica-based stationary phases cause continuous column bleeding and prevent buffer switching during daily usage. , Besides, during heating and ionizing in the ion source, ammonium fluoride will produce hydrofluoric acid, , which may induce erosion to the instrument . Recently, in a pair of progressive contributions, an unexplained decrease (about 50-fold) of sensitivity for estradiol detection has been rigorously reported by Häkkinen’s group after enrolling ammonium fluoride in the mobile phase for 1 year. , …”

Section: Introductionmentioning

confidence: 99%

Rapid Derivatization of Phenolic and Oxime Hydroxyl with Isonicotinoyl Chloride under Aqueous Conditions and Its Application in LC–MS/MS Profiling Multiclass Steroids

et al. 2022

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Quantification of steroids possesses a crucial clinical value in early diagnosis and prognosis evaluation of various endocrine diseases. However, it is still challenging to realize feasible analysis of estrogens, androgens, progestogens, and corticoids within one single workflow. In this study, two derivatization reactions were newly designed for improvement: (1) acylation of phenolic hydroxyl on estrogens with isonicotinoyl chloride (INC) under the catalysis of 4dimethylaminopyridine and (2) post-modification of oxime hydroxyl on hydroxylamine-pretreated ketosteroids with INC. Both reactions could conduct instantaneously at room temperature under aqueous conditions. Moreover, the resulting phenolic-INC and oxime-INC esters exhibited favorable MS responses. Through integrating these derivatization strategies with cold-induced phase separation technology, a feasible LC−MS/MS method was developed for simultaneous quantification of 15 multiclass steroids with proper sample consumption (50 μL serum), satisfying sensitivity (lower limit of quantitation at 0.01−5.00 ng/mL) and high throughput (40 min for samplepreparation). The practical applicability was tested by detecting 30 real samples from pregnant and non-pregnant women. The obtained results showed a good agreement with a previous validated methodology.

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Fully automated method for quantitative determination of steroids in serum: An approach to evaluate steroidogenesis

Cited by 10 publications

References 56 publications

Semi-automated serum steroid profiling with tandem mass spectrometry

Semi-automated serum steroid profiling with tandem mass spectrometry

Dissecting the KNDy hypothesis: KNDy neuron-derived kisspeptins are dispensable for puberty but essential for preserved female fertility and gonadotropin pulsatility

Rapid Derivatization of Phenolic and Oxime Hydroxyl with Isonicotinoyl Chloride under Aqueous Conditions and Its Application in LC–MS/MS Profiling Multiclass Steroids

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