Miniaturization of a chloride ion assay for use in a microtiter format

Merchant, Mark

doi:10.1016/j.microc.2009.01.002

Cited by 7 publications

(10 citation statements)

References 26 publications

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“…The analysis was conducted under ISO 9001:2000 certification. Chloride concentrations were determined spectrophotometrically at 480 nm using a microtitre plate format with ferric nitrate and mercuric thiocyanate, and using sodium chloride as a standard solution (Merchant 2009). …”

Section: Methodsmentioning

confidence: 99%

Effects of hypercapnia on acid–base balance and osmo-/iono-regulation in prawns (Decapoda: Palaemonidae)

Dissanayake

Clough²,

Spicer³

et al. 2010

Aquat. Biol.

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Anthropogenic carbon dioxide-induced 'ocean acidification' is predicted to have major implications for marine organisms. As the oceans becomes increasingly hypercapnic (elevated CO 2 ) and seawater pH decreases, the ability of organisms to maintain extracellular pH homeostasis (acidbase balance) may be compromised. Acid -base regulation occurs by ionic transport, where hydrogen and bicarbonate ions (HCO 3 -) are exchanged for sodium and chloride, respectively (H + /Na + ; HCO 3 -/Cl -), as exemplified by decapod crustaceans. Palaemonid prawns, in particular, are efficient hypo-ionic/osmotic regulators in seawater. We demonstrate that hypercapnic exposure (0.3 kPa) results in short-term (5 to 14 d) extracellular acidosis in 2 efficient ionic/osmo-regulators (thus, acid -base regulators), i.e. Palaemon elegans and P. serratus. Complete hypercapnic compensation was observed in both species after 30 d exposure with no effect on osmotic capacity, but at the expense of extracellular acid -base alteration (alkalosis). Furthermore, the predominantly subtidal species P. serratus was observed to be as tolerant as the intertidal species P. elegans, although 2 differing mechanisms of ionic regulation may be at work, with P. elegans and P. serratus displaying lower and elevated haemolymph ion concentrations (i.e. sodium, chloride and calcium), respectively. KEY WORDS: Hypercapnia · 'Ocean acidification' · Acid -base balance · Osmotic and ionic regulation · PalaemonResale or republication not permitted without written consent of the publisher Aquat Biol 11: 27-36, 2010 The ocean's natural capacity to buffer changes in seawater chemistry caused by CO 2 has been compromised and surface ocean pH has decreased by 0.1 units (since pre-industrial times) as a result of a 30% increase in H + ions (Caldeira & Wickett 2003, Blackford & Gilbert 2007. The reduction in ocean pH is termed 'ocean acidification'. Responsible for this rapid rise in ocean acidity is increasing hypercapnia (elevated levels of dissolved CO 2 ) (which drives the equilibria of Eq. 1 towards the right-hand side). This may have profound effects upon the biota, the critical limits and long-term effects of which are currently unknown (Pörtner et al. 1998, Seibel & Walsh 2003, Widdicombe & Spicer 2008.It is likely that changes in seawater chemistry will affect the internal physiological functioning of marine organisms, such as the acid -base balance (Raven et al. 2005). Organisms that live in aquatic environments face the problem of maintaining a constant internal environment (i.e. acid -base balance in extracellular fluid) that enables cells to function efficiently and is independent of the external environment (Rankin & Davenport 1981, Seibel & Walsh 2003 (Cameron & Mangum 1983, Mantel & Farmer 1983. Ionic and osmotic regulation is defined as 'the maintenance in a body fluid of concentrations of ions (ionic) and total particle concentration (osmotic) differing to that the external medium' (Robertson 1949), and it is generally regarded that efficient ioni...

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Section: Methodsmentioning

confidence: 99%

Effects of hypercapnia on acid–base balance and osmo-/iono-regulation in prawns (Decapoda: Palaemonidae)

Dissanayake

Clough²,

Spicer³

et al. 2010

Aquat. Biol.

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“…Probably the most popular halide assay is the “Iwasaki assay” which has been adapted several times since its introduction in 1952 and has been miniaturised to the microtiter plate format . It is based on the displacement of thiocyanate from mercuric thiocyanate by chloride, followed by the formation of an orange‐red‐coloured ferric thiocyanate complex that can be quantified spectrophotometrically at 460 nm.…”

Section: Resultsmentioning

confidence: 99%

“…For this reason, the Iwasaki assay for halide ions has remained popular in dehalogenase assays, despite the involvement of extremely toxic mercury derivatives . In microtiter plate format, variations of the Iwasaki assay were reported to have detection limits for halide ions between 20 and 270 μM …”

Section: Introductionmentioning

confidence: 99%

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

et al. 2020

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Halide assays are important for the study of enzymatic dehalogenation, a topic of great industrial and scientific importance. Here we describe the development of a very sensitive halide assay that can detect less than a picomole of bromide ions, making it very useful for quantifying enzymatic dehalogenation products. Halides are oxidised under mild conditions using the vanadium‐dependent chloroperoxidase from Curvularia inaequalis, forming hypohalous acids that are detected using aminophenyl fluorescein. The assay is up to three orders of magnitude more sensitive than currently available alternatives, with detection limits of 20 nM for bromide and 1 μM for chloride and iodide. We demonstrate that the assay can be used to determine specific activities of dehalogenases and validate this by comparison to a well‐established GC‐MS method. This new assay will facilitate the identification and characterisation of novel dehalogenases and may also be of interest to those studying other halide‐producing enzymes.

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“…For bromine [54] and chlorine [50] determination few applications using spectrophotometry could be found. A phenol red solution combined with Oxone ® was used by Baso-Cejas et al [53] for the determination of bromide in vegetables after an alkaline ashing procedure.…”

Section: Spectrophotometric Methodsmentioning

confidence: 99%

Analytical methods for the determination of halogens in bioanalytical sciences: a review

et al. 2013

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Fluorine, chlorine, bromine, and iodine have been studied in biological samples and other related matrices owing to the need to understand the biochemical effects in living organisms. In this review, the works published in last 20 years are covered, and the main topics related to sample preparation methods and analytical techniques commonly used for fluorine, chlorine, bromine, and iodine determination in biological samples, food, drugs, and plants used as food or with medical applications are discussed. The commonest sample preparation methods, as extraction and decomposition using combustion and pyrohydrolysis, are reviewed, as well as spectrometric and electroanalytical techniques, spectrophotometry, total reflection X-ray fluorescence, neutron activation analysis, and separation systems using chromatography and electrophoresis. On this aspect, the main analytical challenges and drawbacks are highlighted. A discussion related to the availability of certified reference materials for evaluation of accuracy is also included, as well as a discussion of the official methods used as references for the determination of halogens in the samples covered in this review.

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Miniaturization of a chloride ion assay for use in a microtiter format

Cited by 7 publications

References 26 publications

Effects of hypercapnia on acid–base balance and osmo-/iono-regulation in prawns (Decapoda: Palaemonidae)

Effects of hypercapnia on acid–base balance and osmo-/iono-regulation in prawns (Decapoda: Palaemonidae)

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

Analytical methods for the determination of halogens in bioanalytical sciences: a review

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