Quantification of 2-hydrazinopyridine derivatized steroid hormones in fathead minnow (Pimephales promelas) blood plasma using LC-ESI+/MS/MS

Hala, David; Overturf, Matthew D.; Petersen, Lene H.; Huggett, Duane B.

doi:10.1016/j.jchromb.2011.01.020

Cited by 35 publications

(17 citation statements)

References 62 publications

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“…Norethindrone, ibuprofen, verapamil, and clozapine analytes were extracted from water samples using a liquid-liquid extraction method (1:1 mixture of hexane and ethyl acetate). Dexamethasone, and its associated internal standard, required a derivatization process using a mixture of hydrazinopyridine and trifluoroacetic acid for analysis (Hala et al 2011). All other compounds did not require an extraction process and were analyzed directly from the collected water sample.…”

Section: Methodsmentioning

confidence: 99%

Early Life-Stage Toxicity of Eight Pharmaceuticals to the Fathead Minnow, Pimephales promelas

Overturf

Baxter

et al. 2011

Arch Environ Contam Toxicol

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Human pharmaceuticals are routinely being detected in the environment, and there is growing concern about whether these drugs could elicit effects on aquatic organisms. Regulatory paradigms have shifted accordingly, with a greater emphasis on chronic toxicity data compared with acute data. The Organisation for Economic Co-operation and Development 210 Early Life Stage Test has been proposed as a good measure of the potential for pharmaceuticals to elicit chronic toxicity. To begin building a data set regarding the early life-stage toxicity of pharmaceuticals to fish, fathead minnows (FHM) were exposed to amiodarone, carbamazepine, clozapine, dexamethasone, fenofibrate, ibuprofen, norethindrone, or verapamil. Survival and growth were used to assess chronic toxicity in FHM at 28 days posthatch. Exposure of FHM to carbamazepine, fenofibrate, and ibuprofen resulted in no significant adverse effects at the concentrations tested. FHM survival was not impacted by verapamil exposure; however, growth was significantly decreased at 600 μg/L. Dexamethasone-exposed FHM showed a significant decrease in survival at a concentration of 577 μg/L; however, growth was not impacted at the concentration tested. Norethindrone exposure resulted in a significant decrease in survival and dry weight at 14.8 and 0.74 μg/L, respectively. Exposure to amiodarone and clozapine resulted in a significant decrease in survival and a significant increase in growth at concentrations of 1020 and 30.8 μg/L, respectively. Although the effect levels derived in this study are greater then concentrations observed in the environment, these data suggest that synthetic progestins may require additional research.

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Section: Methodsmentioning

confidence: 99%

Early Life-Stage Toxicity of Eight Pharmaceuticals to the Fathead Minnow, Pimephales promelas

Overturf

Baxter

et al. 2011

Arch Environ Contam Toxicol

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“…From this volume, a 50l aliquot was taken for estrogen quantification using a dansyl chloride derivatization method detailed by Nelson et al (Nelson et al, 2004). The remaining 50l sample was also dried under nitrogen with residue derivatized using 2-hydrazinopyridine as described in Hala et al (Hala et al, 2011). All chromatographic separations were conducted using a Waters 2695 separations module coupled to a Waters 2998 UV/VIS detector and a Waters Sunfire C 18 column (2.1ϫ50mm, 3.5m particle size; Milford, MA, USA).…”

Section: Steroid Hormone Quantification Using Lc-esi+/ms/msmentioning

confidence: 99%

“…However, in the present study, 11-ketotestosterone production was quantified from ovary tissue as it is stoichiometrically related to testosterone catalysis (V39 and 40; Table1), lending a catalytically feasible pathway for its production in female fish. Furthermore, detectable concentrations of 11-ketotestosterone have been quantified in the plasma of adult female fathead minnows (Hala et al, 2011). Moreover, to our knowledge, there are no reported studies investigating the effects of hypoxia on 11-ketotestosterone titers in female fish, whereas studies with male fish show decreased concentrations after long-term (30days) hypoxia exposure (Landry et al, 2007).…”

Section: Effects Of Hypoxia On Steroid Hormone Production -Ex Vivo Anmentioning

confidence: 99%

Constraints-based stoichiometric analysis of hypoxic stress on steroidogenesis in fathead minnows, Pimephales promelas

Hala

Petersen

Martinović

et al. 2012

Journal of Experimental Biology

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SUMMARYIn this study, an in silico genome-scale metabolic model of steroidogenesis was used to investigate the effects of hypoxic stress on steroid hormone productions in fish. Adult female fathead minnows (Pimephales promelas) were exposed to hypoxia for 7days with fish sub-sampled on days 1, 3 and 7 of exposure. At each time point, selected steroid enzyme gene expressions and steroid hormone productions were quantified in ovaries. Fold changes in steroid enzyme gene expressions were used to qualitatively scale transcript enzyme reaction constraints (akin to the range of an enzymeʼs catalytic activity) in the in silico model. Subsequently, in silico predicted steroid hormone productions were qualitatively compared with experimental results. Key findings were as follows. (1) In silico gene deletion analysis identified highly conserved ʻessentialʼ genes required for steroid hormone productions. These agreed well (75%) with literature-published genes downregulated in vertebrates (fish and mammal) exposed to hypoxia. (2) Quantification of steroid hormones produced ex vivo from ovaries showed a significant reduction for 17-estradiol and 17,20-dihydroxypregnenone production after 24h (day 1) of exposure. This lowered 17-estradiol production was concomitant with downregulation of cyp19a1a gene expression in ovaries. In silico predictions showed agreement with experimentation by predicting effects on estrogen (17-estradiol and estrone) production. (3) Stochastic sampling of in silico reactions indicated that cholesterol uptake and catalysis to pregnenolone along with estrogen methyltransferase and glucuronidation reactions were also impacted by hypoxia. Taken together, this in silico analysis introduces a powerful model for pathway analysis that can lend insights on the effects of various stressor scenarios on metabolic functions. Supplementary material available online at

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“…Levonorgestrel was extracted from water samples using a liquid-liquid extraction method (1:1 mixture of hexane and ethyl acetate). Both LNG and d6-NOR required a derivatization process using a mixture of hydrazinopyridine and trifluoroacetic acid (HP/TFA) for analysis (Hala et al, 2011).…”

Section: Liquid Chromatography-tandem Mass Spectrometry (Lc-ms/ms)mentioning

confidence: 99%

Responses to various exposure durations of levonorgestrel during early-life stages of fathead minnows (Pimephales promelas)

Overturf

Huggett

2015

Aquatic Toxicology

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Quantification of 2-hydrazinopyridine derivatized steroid hormones in fathead minnow (Pimephales promelas) blood plasma using LC-ESI+/MS/MS

Cited by 35 publications

References 62 publications

Early Life-Stage Toxicity of Eight Pharmaceuticals to the Fathead Minnow, Pimephales promelas

Early Life-Stage Toxicity of Eight Pharmaceuticals to the Fathead Minnow, Pimephales promelas

Constraints-based stoichiometric analysis of hypoxic stress on steroidogenesis in fathead minnows, Pimephales promelas

Responses to various exposure durations of levonorgestrel during early-life stages of fathead minnows (Pimephales promelas)

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