Direct detection and quantification of 19-norandrosterone sulfate in human urine by liquid chromatography-linear ion trap mass spectrometry

Strahm, Emmanuel; Saudan, C; Sottas, Pierre-Edouard; Mangin, Patrice; Saugy, Martial

doi:10.1016/j.jchromb.2007.02.009

Cited by 17 publications

(10 citation statements)

References 27 publications

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“…Similarly to previous studies [9,14], [15][16][17][18]. The ion at m/z 97 corresponding to HSO 4 − is observed when the sulfate group is attached to an alicyclic ring and appears to originate from hydrogen abstraction at position 3 of the steroid moiety [17,19].…”

Section: Chromatography and Mass Spectrometrysupporting

confidence: 77%

“…Referenced protocols authorizing the separation of glucuro and sulfoconjugates of steroids/corticosteroids are time consuming, and in addition the use of three SPE cartridges to isolate sulfate species for each urine specimen is required [9,25]. The sample preparation protocol used in this work was performed on a single WAX SPE cartridge for selective extraction of 19-NE and 19-NA conjugates.…”

Section: Ms/ms Spectrum Of the Deuterated Internal Standard D 4 -mentioning

confidence: 99%

“…For comparison purpose, it is noteworthy that referenced protocols based on the indirect quantification of 19-NAS and 19-NES levels presented lower quantitative performance in terms of sensitivity and range of quantification [12]. Regarding the use of LC/MS/MS for direct quantification of sulfoconjugates, it has been shown that assessment of 19-NAS at subnanogram mL −1 level could be performed using a SPE multi-step approach [9]. Although the matrix effect in LC/MS/MS was significantly reduced after intensive purification, the procedure required the sequential use of three SPE cartridges to isolate sulfate species for each urine specimen (1 day-extraction).…”

Section: Assay Validationmentioning

confidence: 99%

“…In the case of sulfoconjugates, this analytical tool is of great interest as it does not require hydrolysis and derivatisation of the analytes [8]. A first method for direct quantification of 19-norandrosterone sulfate in human urine has been recently published [9]. In the work present herein, a fast method for a selective separation of glucuronide and sulfate using weak anion exchange solid phase extraction support was developed.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Profiling of 19-norsteroid sulfoconjugates in human urine by liquid chromatography mass spectrometry

Strahm¹,

Rudaz

Veuthey

et al. 2008

Analytica Chimica Acta

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Section: Chromatography and Mass Spectrometrysupporting

confidence: 77%

Section: Ms/ms Spectrum Of the Deuterated Internal Standard D 4 -mentioning

confidence: 99%

Section: Assay Validationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Profiling of 19-norsteroid sulfoconjugates in human urine by liquid chromatography mass spectrometry

Strahm¹,

Rudaz

Veuthey

et al. 2008

Analytica Chimica Acta

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“…In this study, we followed the method described by Strahm et al [26], with significant modifications. Briefly, 10 mL of urine sample was centrifuged at 3500 r/min for 5 min, and a volume of 10 mL acetate buffer (pH 5.2, 0.2 mol/L) was added into the supernatant.…”

Section: Lc-ms/ms Determinationmentioning

confidence: 99%

Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue

et al. 2011

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has been illegally used in horse racing to boost physical performance, and in animal husbandry to accelerate weight gain. To monitor the abuse of NT, our goal was to develop a commercial enzyme linked immunosorbent assay (ELISA) kit. For this purpose, hybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mouse. Noncompetitive and competitive indirect ELISA were used to screen positive cell clones. To optimize the indirect competitive ELISA (icELISA) method, various methanol concentrations in assay buffer were evaluated. Matrix effects in urine and spiking test were also investigated. Finally, five hybridoma cell lines named NT-1, NT-2, NT-3, NT-4 and NT-5 were screened out. The corresponding monoclonal antibodies (mAbs) were of the IgG 1 isotype with a k light chain, and the antibody affinity of all mAbs were between 2.6×10 9 and 4.7×10 9 L/mol. The titer and IC 50 values of purified ascites were in the range of 0.64×10 5 −2.56×10 5 and 0.55-1.0 ng/mL, respectively. Based on the NT-1 hybridoma, a heterologous icELISA method was developed for the quantitative detection of NT in cattle urine. The dynamic range was from 0.004 to 85.8 ng/mL, with a detection limit for the assay and IC 50 values of 0.002 and 0.55 ng/mL, respectively. Except for a high cross-reactivity (62%) to α-NT, negligible cross-reactivity to other compounds was observed. After optimization, 10% of methanol was used in the assay buffer, and a 20-fold dilution in cattle urine gave an inhibition curve almost the same as that in phosphate buffered saline. The correlation coefficient between the established icELISA and LC-MS/MS method was 0.9871. The results showed that the established heterologous icELISA method provides an excellent alternative for the detection of NT residues in food producing animals. 19-nortestosterone

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Simultaneous quantitation and identification of intact Nandrolone phase II oxo‐metabolites based on derivatization and inject LC–MS/(HRMS) methodology

Sakellariou,

Kiousi,

Petrou

et al. 2024

Drug Testing and Analysis

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AbstractΑ sensitive and selective derivatization and inject method for the quantification of intact nandrolone phase II oxo‐metabolites was developed and validated using liquid chromatography ‐ (tandem high resolution) mass spectrometry (LC–MS/(HRMS)). For the derivatization, Girard's reagent T (GRT) was used directly in natural urine samples and the analysis of the metabolites of interest was performed by direct injection into LC–MS/(HRMS) system operating in positive ionization mode. Derivatization enabled the efficient detection of nandrolone oxo‐metabolites, while at the same time producing intense product ions under collision‐induced dissociation (CID) conditions that are related to metabolites of the steroid backbone and not to the conjugated moieties. Glucuronide and sulfate metabolites of nandrolone were chromatographically resolved and quantified in the same run in the range of 1–100 ng mL−1, while at the same time structure identification could be performed for each metabolite. Full validation of the method was performed according to the World Anti‐Doping Agency (WADA) International Standard for Laboratories (ISL). Nandrolone oxo‐metabolites were quantified in two sets of urine samples, the first set consisted of real urine samples previously detected as negative and the second set consisted of urine samples collected from two excretion studies after nandrolone decanoate administration. The results for 19‐norandrosterone glucuronide (19‐NAG) and 19‐noretiocholanolone glucuronide (19‐NEG) were compared with those obtained by traditional gas chromatography ‐ (tandem) mass spectrometry (GC–MS/[MS]) method.

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Direct detection and quantification of 19-norandrosterone sulfate in human urine by liquid chromatography-linear ion trap mass spectrometry

Cited by 17 publications

References 27 publications

Profiling of 19-norsteroid sulfoconjugates in human urine by liquid chromatography mass spectrometry

Profiling of 19-norsteroid sulfoconjugates in human urine by liquid chromatography mass spectrometry

Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue

Simultaneous quantitation and identification of intact Nandrolone phase II oxo‐metabolites based on derivatization and inject LC–MS/(HRMS) methodology

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