Repeatability of the Submitochondrial Particle Assay

Doherty, Francis G.; Gustavson, Karl E.

doi:10.1006/eesa.2002.2245

Cited by 5 publications

(6 citation statements)

References 19 publications

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“…Doherty and Gustavson [16] assessed the toxicities of sodium dodecylsulfate, pentachlorophenol, and zinc sulfate in both the RET and ETr variants to define the repeatability or within-laboratory level of variability associated with repeated trials similar to that required in the generation of quality control charts. Efforts to further develop the fundamental design of the assays in order to standardize their conductance have been neglected.…”

Section: Discussionmentioning

confidence: 99%

“…The test endpoint is the loss of nicotinamide adenine dinucleotide (NADH) sodium salt, reduced form, monitored at 340 nm in a standard spectrophotometer using 1 cm path length disposable methyl acrylate cuvettes. Generalized descriptions of the conductance of both assays are provided elsewhere [16]. In this test, electrons donated by succinate at Complex II are unable to progress down the electron transfer chain because antimycin A blocks electron flow through Complex III.…”

Section: Electron Transfer and Reverse Electron Transfer Assaysmentioning

confidence: 99%

“…Energy derived from the hydrolysis of adenosine-5-triphosphate drives electrons in the reverse direction, up the electron transport chain to Complex I, where NAD ϩ is reduced to NADH, again monitored at 340 nm on a standard spectrophotometer [15]. Generalized descriptions of the conductance of both assays are provided elsewhere [16].…”

Section: Electron Transfer and Reverse Electron Transfer Assaysmentioning

confidence: 99%

“…Demonstration of assay reproducibility is limited to a single study assessing within-laboratory repeatability [16]. Manual test protocols only include an approach for generating test endpoints using serial dilution of test solutions.…”

Section: Introductionmentioning

confidence: 99%

“…The data analysis approach currently in use involves a partially subjective selection of data points. Demonstration of assay reproducibility is limited to a single study assessing within-laboratory repeatability [16]. In addition, SMPs are a manufactured test system.…”

Section: Introductionmentioning

confidence: 99%

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Effect of storage temperature on the activity of submitochondrial particles

Doherty¹

2008

Enviro Toxic and Chemistry

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The submitochondrial particle (SMP) assay employs processed mammalian mitchondria to assess the toxicity of chemical contaminants in aqueous solutions. Particles and associated reagents are commercially available to support two individual procedures, the electron transfer (ETr) and reverse electron transfer (RET) assays. The objective of the present study was to assess the effect of storage temperature on SMP activity. One RET and one ETr assay were conducted with sodium dodecylsulfate on each of two vials of particles stored at -20 and -80 degrees C at periodic intervals over a six-month span. Results demonstrated that SMP could remain active in either assay through six months of storage at either temperature. However, there were isolated vials of particles stored at -20 degrees C that exhibited unacceptable reductions in activity for both the ETr and the RET assays that were not related to storage duration. These results were used to develop guidance in assessing the acceptability of particle activity in SMP assays.

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Section: Discussionmentioning

confidence: 99%

Section: Electron Transfer and Reverse Electron Transfer Assaysmentioning

confidence: 99%

Section: Electron Transfer and Reverse Electron Transfer Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effect of storage temperature on the activity of submitochondrial particles

Doherty¹

2008

Enviro Toxic and Chemistry

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The Submitochondrial Particle Assay as a Biological Monitoring Tool

Doherty¹

2004

Water Encyclopedia

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While studies assessing the toxicity of environmental contaminants to aquatic organisms date back to the early 1900s, the use of toxicity testing as a tool for monitoring the environmental acceptability of discharges from wastewater treatment and manufacturing facilities to surface waters (i.e., rivers, lakes, and oceans) did not begin to evolve until the 1970s. One of the primary motivating factors behind the development of biological monitoring, or biomonitoring, was the manufacture of exponentially increasing numbers of new chemicals for which analytical procedures were generally lacking. Biomonitoring was viewed as an approach to detect the presence of potentially toxic chemicals in facility discharges without necessarily having to identify the contaminants at that stage of the assessment. Additional advantages included the reduced expense associated with biologically testing significantly greater numbers of samples in comparison with analytical procedures. This approach also focused on only biologically available forms of the chemicals in solution reflecting any interactive effects (synergistic or antagonistic) that might be occurring among contaminants within a complex sample.

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