Interference of the Schlieren effect on sea water analysis by spectrophotometry is caused by the flow of solutions of different ionic strengths through a flow cell. A flow injection assay protocol programmed in a flow-batch format removes this interference and allows the use of a calibration line, obtained in deionized water, for determination of analytes in sea water samples of different salinity. This Single Line Calibration (SLC) technique is validated on the most frequently performed nutrient assays.
View Article and Find Full Text PDFThe sensitivity of a spectrophotometric assay is enhanced either by increasing the concentration of the target molecules within the flow cell or by extending the length of the light path of the flow cell. Determination of nutrients at nanomolar concentrations in sea water has therefore been based either on the preconcentration of the analyte on a microcolumn from a large volume of sample followed by its elution into a conventional 1-2 cm flow cell, or by the use of Liquid Core Waveguide (LCW) with a light path as long as several meters. In order to evaluate the relative improvements of these different approaches to increasing sensitivity we have developed a preconcentration technique for the determination of nitrite in seawater using the Gries reaction and compare its sensitivity and precision to that of non preconcentration techniques using both LCW and Linear Light Path (LLP) cells of different lengths.
View Article and Find Full Text PDFThe ultimate goal of flow-based analytical techniques is to automate serial assays of a target analyte. However, when developing any reagent-based assay, the underlying chemistry has to be investigated and understood a step, which is almost always the most challenging component of the optimization effort. The difficulty lies in that almost all reagent-based assays were initially developed and optimized in a batch mode, with the aim to perform assays manually, within a time frame of up to 15 min, while flow injection techniques are designed to monitor concentration gradients at times prior to reaching chemical equilibria and while performing up to two assays per minute.
View Article and Find Full Text PDFAn assay protocol, based on programmable Flow Injection (pFI), is optimized by tailoring flowrates appropriately to the individual steps of an assay, thus allowing sample and reagent metering, mixing, incubation, monitoring and washout to be carried out more efficiently and in different time frames. This novel approach to flow based methods is applied here to optimize the determination of orthophosphate at nanomolar levels. Programmable Flow Injection was also used to facilitate an investigation of the properties of the phosphomolybdenum blue (PMoB) formed during this assay, by using the stop flow technique - an approach that revealed for the first time the influence of surfactants on the kinetics of formation of PMoB and its spectral characteristics.
View Article and Find Full Text PDFAutomation of reagent based assays by Flow Injection is based on sample processing, in which a sample flows continuously towards and through a detector for monitoring of its components. There are three drawbacks to using this approach. The constant continuous forward flow: continually consumes reagents and generates chemical waste and necessitates a compromise when optimizing the performance of the reagent based assay.
View Article and Find Full Text PDFBy virtue of their compactness, long-term stability, minimal reagent consumption and robustness, miniaturized sequential injection instruments are well suited for automation of assays onboard research ships. However, in order to reach the sensitivity and limit of detection required for open-ocean determinations of trace elements, it is necessary to preconcentrate the analyte prior its derivatization and subsequent detection by fluorescence. In this work, a novel method for the determination of dissolved zinc (Zn) at subnanomolar levels in seawater is described.
View Article and Find Full Text PDFMicro-sequential injection lab-on-valve (µSI-LOV) is a well-established analytical platform for absorbance and fluorescence based assays but its applicability to chemiluminescence detection remains largely unexplored. In this work, we describe a novel fluidic protocol and two distinct strategies for photon collection that enable chemiluminescence detection using µSI-LOV for the first time. To illustrate this proof of concept, we selected the reaction between Fe(II) and luminol and developed a preliminary protocol for Fe(II) determinations in acidified seawater.
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