Comparison of Unit Resolution Srm And Tof-Ms At 12,000 Mass Resolution For Quantitative Bioanalysis of 11 Steroids From Human Plasma

Rousu, Timo; Tolonen, Ari

doi:10.4155/bio.11.289

Cited by 16 publications

(7 citation statements)

References 20 publications

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“…The LOD for the individual analyte was determined at the lowest concentration, showing a signal to noise ratio (S/N) superior to 3, and ranged from 0.1 to 0.5 ng/mL (Table 2). The LOQ for the individual analyte was determined at the lowest concentration, showing a signal to noise ratio (S/N) superior to 10, and ranged from 0.3 to 1.7 ng/mL (Table 2), similar to the respective values reported recently [41]. 1.…”

Section: Calibration and Methods Validationsupporting

confidence: 82%

“…In contrast with QQQ MS systems, the TOF-MS systems have been traditionally used for screening-type analyses rather than fully quantitative work. Recently, TOF-MS technology has gained the advantage of increased applicability for quantitative work, by the increase in the linear (dynamic) range and mass resolution of the instruments [41]. The most used Q-TOF or Orbitrap mass system possess a dynamic range of ≥4 orders of magnitude, a mass resolution of ≥30,000 (full width at half maximum), and a mass accuracy of <5 ppm.…”

Section: Methods Developmentmentioning

confidence: 99%

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UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

Zhi

et al. 2021

Molecules

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The quantification of steroid hormones of individual zebrafish (Danio rerio) provides perspective to understand endogenous hormone function. A UPLC–TOF–MS method was developed to provide a reproducible, sensitive, and efficient assay to determine the concentration of steroid hormones, including cortisol, testosterone, androstenedione, 11-deoxycortisol, 11-deoxycorticosterone, and 17-hydroxyprogesterone in whole-body homogenates of each zebrafish. Solid-phase extraction was used to sample matrix clean-up and acquired a recovery from 89.7% to 107.9%. The analytes were separated on an Aquity BEH C18 column using gradient elution. Mass spectrometric analysis was performed by single reaction monitoring (SRM) using positive electrospray ionization mode. The total running time was 6 min, which was greatly shortened compared with a previously reported method. The developed method exhibited excellent linearity for all the analytes, with regression coefficients higher than 0.99. The limit of detection varied between 0.1 and 0.5 ng/L and the limit of quantification was 0.5–1.7 ng/L for all analytes. The precision of the method was assessed on replicate measurements and was found to be in the ranges of 1.9 % to 6.6% and 4.3% to 8.6%, for intra- and inter-day analysis, respectively. This method was validated according to FDA guidance and applied to determine steroid hormone levels in the tissue homogenate of zebrafish acutely treated with caffeine and ethanol.

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Section: Calibration and Methods Validationsupporting

confidence: 82%

Section: Methods Developmentmentioning

confidence: 99%

UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

Zhi

et al. 2021

Molecules

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“…The LLE method is appreciated for its speed and simplicity, while the SPE method can be semi-automated using 96well plates for instance [106]. The quality of the extraction technique used is evaluated by the calculation of the percentage of steroid recovery (% of recovery), precision, and accuracy as well as the matrix effect (%) [108].…”

Section: Methodologiesmentioning

confidence: 99%

Six Decades of Research on Human Fetal Gonadal Steroids

Connan-Perrot

Léger

Lelandais

et al. 2021

IJMS

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Human fetal gonads acquire endocrine steroidogenic capabilities early during their differentiation. Genetic studies show that this endocrine function plays a central role in the sexually dimorphic development of the external genitalia during fetal development. When this endocrine function is dysregulated, congenital malformations and pathologies are the result. In this review, we explain how the current knowledge of steroidogenesis in human fetal gonads has benefited from both the technological advances in steroid measurements and the assembly of detailed knowledge of steroidogenesis machinery and its expression in human fetal gonads. We summarise how the conversion of radiolabelled steroid precursors, antibody-based assays, mass spectrometry, ultrastructural studies, and the in situ labelling of proteins and mRNA have all provided complementary information. In this review, our discussion goes beyond the debate on recommendations concerning the best choice between the different available technologies, and their degrees of reproducibility and sensitivity. The available technologies and techniques can be used for different purposes and, as long as all quality controls are rigorously employed, the question is how to maximise the generation of robust, reproducible data on steroid hormones and their crucial roles in human fetal development and subsequent functions.

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“…For most analytes, the ULOQ was 400–1000 nM, as indicated by detector saturation in the calibration runs when exceeding this concentration. This low linear range is a typical limitation of time-of-flight analyzers [ 59 ]. The IM stage results in a further accumulation of analytes due to the trapping of ions upstream of the drift tube, which leads to even more pronounced saturation effects.…”

Section: Discussionmentioning

confidence: 99%

Non-targeted and targeted analysis of oxylipins in combination with charge-switch derivatization by ion mobility high-resolution mass spectrometry

et al. 2020

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Eicosanoids and other oxylipins play an important role in mediating inflammation as well as other biological processes. For the investigation of their biological role(s), comprehensive analytical methods are necessary, which are able to provide reliable identification and quantification of these compounds in biological matrices. Using charge-switch derivatization with AMPP ( N -(4-aminomethylphenyl)pyridinium chloride) in combination with liquid chromatography ion mobility quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS), we developed a non-target approach to analyze oxylipins in plasma, serum, and cells. The developed workflow makes use of an ion mobility resolved fragmentation to pinpoint derivatized molecules based on the cleavage of AMPP, which yields two specific fragment ions. This allows a reliable identification of known and unknown eicosanoids and other oxylipins. We characterized the workflow using 52 different oxylipins and investigated their fragmentation patterns and ion mobilities. Limits of detection ranged between 0.2 and 10.0 nM (1.0–50 pg on column), which is comparable with other state-of-the-art methods using LC triple quadrupole (QqQ) MS. Moreover, we applied this strategy to analyze oxylipins in different biologically relevant matrices, as cultured cells, human plasma, and serum. Graphical abstract Electronic supplementary material The online version of this article (10.1007/s00216-020-02795-2) contains supplementary material, which is available to authorized users.

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Comparison of Unit Resolution Srm And Tof-Ms At 12,000 Mass Resolution For Quantitative Bioanalysis of 11 Steroids From Human Plasma

Cited by 16 publications

References 20 publications

UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

Six Decades of Research on Human Fetal Gonadal Steroids

Non-targeted and targeted analysis of oxylipins in combination with charge-switch derivatization by ion mobility high-resolution mass spectrometry

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