Colorimetric determination of nanomolar concentrations of ammonium in seawater using solvent extraction

A solvent extraction method for measuring nanomolar concentrations of ammonium in seawater is described. The procedure is based on formation of indophenol in alkaline solution by reaction of phenol, hypochlorite and ammonia using sodium aquopentacyanoferrate as a coupling agent. Indophenol is then concentrated by extraction into n-hexanol at low pH and re-extraction into aqueous alkaline buffer. The concentration of indophenol blue is determined colorimetrically. The molar absorbance is 2.08 × 10⁵ absorbance units per molar NH₄⁺ in seawater with a precision of ± 1.9 nM NH₄⁺ (95% Cl) for concentrations ≤ 50 nanomolar. This represents a 12-fold improvement in sensitivity and a 26-fold improvement in precision over standard aqueous analyses. Calibration curves are linear to at least 2 μM NH₄⁺. Sensitivity in seawater is 97% of that found in deionized distilled water due to a slight salt effect.     

Determination of ammonium regeneration rates in the oligotrophic ocean by gas chromatography/mass spectrometry

A method has been developed for the determination of ammonium concentration and isotopic enrichment in seawater samples at the low nanomolar range (10–100 nmol/kg). It is based on the reaction of phenol/hypochlorite with ammonium to form indophenol, with subsequent solid phase extraction, derivatisation and analysis by Gas Chromatography Mass Spectrometry. The precision of the method was maximised by incorporating a deuterated indophenol internal standard. A system was developed which generated seawater with extremely low ammonium concentrations thus matching sample and standard matrices for quantitative analysis. Data are presented from a study of ammonium regeneration rates at three stations in the oligotrophic North–East Atlantic where ambient ammonium concentrations were < 21 nmol/kg. Results suggested that ammonium availability for phytoplankton was limited by the rate of ammonium regeneration. Efficient ammonium assimilation contributed to the very low ambient ammonium concentrations measured at these stations. The study highlights the need for the accurate determination of ammonium regeneration rates in studies of new production, particularly in extreme oligotrophic conditions. If not corrected for isotope dilution, f-ratio estimates may be overestimated by 10.7–13.7%.     

A solvent extraction method for the colorimetric determination of nanomolar concentrations of silicic acid in seawater

A solvent extraction method for measuring nanomolar concentrations of silicic acid in seawater is described. The procedure is based on the formation of beta silicomolybdic acid by reaction of silicic and molybdic acids at low pH, extraction of the combined acid into n-butanol and reduction with a mixture of p-methylaminophenol sulfate and sulfite. The concentration of the resulting blue silicomolybdous acid in the extract is determined colorimetrically. The method has 30 times the sensitivity and 14 times the precision of standard aqueous analyses. Molar absorbance is 2.29 × 10⁵ in seawater with a precision of ± 2.5 nM Si for concentrations <- 50 nanomolar. Sensitivity in seawater is 70% of that in deionized distilled water owing to a significant salt effect. Natural concentrations of arsenate, arsenite and germanic acid cause negligible interference; however, phosphate interference is equivalent to 11 ± 1 nM Si over a broad range of phosphate concentrations, resulting in an error of ± 1 nM in the corrected silicic acid concentration measurement.     

海水油荧光测量方法实验研究

根据荧光测量方法的原理,采和双光路,双通道及单片微机控制,数据自动采集的技术,于１９９７年研制成功用于海洋现场探测的单波段水中油荧光计。应用该水中油荧光计进行了海水中微量油份的测量实验研究。     

Liquid waveguide spectrophotometric measurements of arsenate and particulate arsenic,as well as phosphate and particulate phosphorus,in seawater

Sensitive methods for the determination of arsenate and particulate arsenic (PAs), as well as phosphate and particulate phosphorus (PP), in seawater are described. The method for arsenate and phosphate was established by applying automated liquid waveguide spectrophotometry. Because the reaction time for the formation of the arsenate-molybdate complex is longer than that for the phosphate-molybdate complex, a long Teflon tube submerged in a heating bath was installed in the conventional phosphate flow system. The arsenate was quantified as the difference between absorbances of molybdenum blue dyes with (only phosphate) and without (phosphate + arsenate) arsenate reduction treatment. Contamination was observed in the reagent for arsenate reduction and must be corrected. Linear dynamic ranges up to 1000 nM were confirmed for arsenate and phosphate. The detection limits for arsenate and phosphate were 5 and 4 nM, respectively. Freezing was a reliable sample preservation technique for both arsenate and phosphate. The method for PAs and PP was established by combining conventional persulfate oxidation of PP and the automated liquid waveguide spectrophotometry of arsenate and phosphate. The digestion efficiencies of organic As analogs were >93%. Contamination in the glass fiber filter was negligible. Field tests confirmed that the coefficients of variation (CVs) of 10–19 nM arsenate and 4–151 nM phosphate were 7–20 and 1–25%, respectively, while the CVs of 0.9 nM PAs and 10.2 nM PP were 11 and 4%, respectively.     

Photochemical formation of glyoxylic and pyruvic acids in seawater

Glyoxylic and pyruvic acids were formed when filter-sterilized seawater was exposed to solar radiation. Production rates varied from samples collected from distinctly different regions of the sea. Humic-rich seawater from the Florida Bay exhibited net photochemical production rates (glyoxylate, 27.5 nM/W-h m⁻²; pyruvate, 12.9 nM/W-h m⁻²) that were significantly greater than net production rates for humic-poor water (glyoxylate, 3.1 nM/W-h m⁻²; pyruvate, 1.8 nM/W-h m⁻²) collected in the Gulf Stream. When seawater was not filtered, the concentrations of glyoxylate and pyruvate were found to undergo diurnal variations resulting from an imbalance between biological and photochemical processes.A depth profile of the glyoxylate concentration from several oceanic stations showed a pronounced daytime maximum in the upper 10 m; this finding is consistent with laboratory results that demonstrated that glyoxylate is formed photochemically in seawater. Pyruvate, in contrast, showed no clear trend with depth; its distribution in the water column may be primarily controlled by biological processes rather than by photochemical processes.Biological processes are generally thought to control the spatial and temporal distribution of simple organic metabolites in seawater. Our results show that photochemical processes may also be important in the marine cycling of some biochemical compounds.     

Impact of environmental factors on in situ determination of iron in seawater by flow injection analysis

A sensitive method for iron determination in seawater has been adapted on a submersible chemical analyser for in situ measurements. The technique is based on flow injection analysis (FIA) coupled with spectrophotometric detection. When direct injection of seawater was used, the detection limit was 1.6 nM, and the precision 7%, for a triplicate injection of a 4 nM standard. At low iron concentrations, on line preconcentration using a column filled with 8-hydroxyquinoline (8HQ) resin was used. The detection limit was 0.15 nM (time of preconcentration = 240 s), and the precision 6%, for a triplicate determination of a 1 nM standard, allowing the determination of Fe in most of the oceanic regimes, except the most depleted surface waters. The effect of temperature, pressure, salinity, copper, manganese, and iron speciation on the response of the analyser was investigated. The slope of the calibration curves followed a linear relation as a function of pressure (C_p = 2.8 × 10^{− 5}P + 3.4 × 10^{− 2} s nmol^{− 1}, R² = 0.997, for Θ = 13 °C) and an exponential relation as a function of temperature (C_Θ = 0.009e^0.103Θ, R² = 0.832, for P = 3 bar). No statistical difference at 95% confidence level was observed for samples of different salinities (S = 0, 20, 35). Only very high concentration of copper (1000 × [Fe]) produced a detectable interference. The chemical analyser was deployed in the coastal environment of the Bay of Brest to investigate the effect of iron speciation on the response of the analyser. Direct injection was used and seawater samples were acidified on line for 80 s. Dissolved iron (DFe, filtered seawater (0.4 μm), acidified and stored at pH 1.8) corresponded to 29 ± 4% of Fe_a (unfiltered seawater, acidified in line at pH 1.8 for 80 s). Most of Fe_a (71 ± 4%) was probably a fraction of total dissolvable iron (TDFe, unfiltered seawater, acidified and stored at pH 1.8).     

Reactions which remove dissolved alumina from seawater

Dissolved alumina can coprecipitate with dissolved silica from seawater enriched with both compounds. This coprecipitation is almost complete within 1h and maintains the concentration of dissolved alumina near naturally occurring oceanic concentrations, well below the alumina concentration of 0.50 ppm A1 found to be stable in filtered Sargasso seawater at 2°C. Only 0.5 ppm Si is necessary to initiate this coprecipitation, which indicates that the concentrations of dissolved alumina that occur in seawater and in interstitial water are a function of the concentration of dissolved silica as well as of the alumina solubility.Dissolved alumina is also quickly removed by solid amorphous silica from solutions of seawater enriched in dissolved alumina and also by several marine sediments in contact with the solution. This process may be an important factor in authigenic mineral formation in marine sediments.     

Simple determination of dimethylsulfide (DMS) and dimethylsulfoniopropionate (DMSP) using solid-phase microextraction and gas chromatography-mass spectrometry

Solid-phase microextraction (SPME) is a simple, sensitive and less destructive method for the determination of dimethylsulfide (DMS) in seawater. Combined with detection by gas chromatography-mass spectrometry (GC-MS), the method had sufficient sensitivity (minimum detectable concentration of DMS was 0.05 nM), and practical levels of reproducibility (relative standard deviation ≤7%) and linearity (r ² > 0.995) over a wide concentration range (0.5 to 910 nM). The protocol developed was applied to a Sagami Bay water sample to determine concentrations of DMS and DMSP, and in situ DMSP-lyase activity.     

Mercury concentration in the ocean

Seventy percent of 342 seawater samples collected in the Bering Sea, North and South Pacific, Japan Sea, East and South China Seas, and Indian Ocean had concentrations of “total” mercury ranging from 3 to 6 ng Hg l^?1 with an arithmetic mean of 5.3 ng l^?1 and a geometric mean of 5.0 ng l^?1. In some cases, a higher concentration was observed at the surface, at the halocline or thermocline, or in the bottom water. But in general, there was no consistent correlation between mercury concentration and depth, except for a statistical tendency for mercury concentration to be slightly higher in the surface water. This tendency suggests that mercury in the ocean is supplied from the atmosphere by rain washout. The latitudinal variation of surface mercury concentrations showed that the maximum concentration at each latitude decreased from 40°N to 30°S. This variation provides evidence that atmospheric mercury is emitted mainly from continental areas naturally or anthropogenically.     

Dimethylsulphide and dimethylsulphoniopropionate in Antarctic sea ice and their release during sea ice melting

This study presents concentrations of dimethylsulphide (DMS) and its precursor compound dimethylsulphoniopropionate (DMSP) in a variety of sea ice and seawater habitats in the Antarctic Sea Ice Zone (ASIZ) during spring and summer. Sixty-two sea ice cores of pack and fast ice were collected from twenty-seven sites across an area of the eastern ASIZ (64°E to 110°E; and the Antarctic coastline north to 62°S). Concentrations of DMS in 81 sections of sea ice ranged from < 0.3 to 75 nM, with an average of 12 nM. DMSP in 60 whole sea ice cores ranged from 25 to 796 nM and showed a negative relationship with ice thickness (y = 125x^− 0.8). Extremely high DMSP concentrations were found in 2 cores of rafted sea ice (2910 and 1110 nM). The relationship of DMSP with ice thickness (excluding rafted ice) suggests that the release of large amounts of DMSP during sea ice melting may occur in discrete areas defined by ice thickness distribution, and may produce ‘hot spots’ of elevated seawater DMS concentration of the order of 100 nM. During early summer across a 500 km transect through melting pack ice, elevated DMS concentrations (range 21–37 nM, mean 31 nM, n = 15) were found in surface seawater. This band of elevated DMS concentration appeared to have been associated with the release of sea ice DMS and DMSP rather than in situ production by an ice edge algal bloom, as chlorophyll a concentrations were relatively low (0.09–0.42 μg l^− 1). During fast ice melting in the area of Davis station, Prydz Bay, sea ice DMSP was released mostly as extracellular DMSP, since intracellular DMSP was negligible in both hyposaline brine (5 ppt) and in a melt water lens (4–5 ppt), while extracellular DMSP concentrations were as high as 149 and 54 nM, respectively in these habitats. DMS in a melt water lens was relatively high at 11 nM. During the ice-free summer in the coastal Davis area, DMS concentrations in surface seawater were highest immediately following breakout of the fast ice cover in late December (range 5–14 nM), and then remained at relatively low concentrations through to late February (< 0.3–6 nM). These measurements support the view that the melting of Antarctic sea ice produces elevated seawater DMS due to release of sea ice DMS and DMSP.     

A modified analytical method for the shipboard determination of nanomolar concentrations of orthophosphate in seawater

A method for the determination of nanomolar concentrations of orthophosphate in oligotrophic seawater developed by Liang et al. (2007) has been modified to make it fully feasible for shipboard application and for faster sample throughput with minimized sample volume. The technique is based on the flow injection method with solid phase extraction on a Sep-Pak C18 cartridge and colorimetric detector. The Schlieren effect was minimized by rinsing the cartridge sequentially with 5 mL water and 2 mL 95% ethanol solution. With three micro pumps in parallel, savings of up to 80% in amount of reagents and 25% volume of seawater samples could be achieved in comparison to the previous method. Variation of stopped flow time and sample loading time gave 3 different standard curves, which corresponded to 3 linear ranges within 3.4 and 515 nM. The modified method permits the analysis of samples over a wide range of concentrations, and has been successfully applied to shipboard determination of trace orthophosphate in more than 200 seawater samples during a one-month cruise in the South China Sea. For seawater at concentrations of 20.6, 82.5, 206.2 nM orthophosphate, the relative standard deviations (RSD) (n = 6), determined daily for 6 days on board ship were 4.45%, 4.73% and 6.75%, respectively. Five seawater samples collected in the Station SEATS (South East Asia Time Series Station at 18°N, 116°E) were analyzed using the present method both on board and in a land-based laboratory, as well as with the magnesium hydroxide-induced coprecipitation (MAGIC) method, and showed no significant difference according to the statistical t-test.     

The distribution and preferential biological uptake of cadmium at 6°W in the Southern Ocean

The spatial and temporal distribution of cadmium (Cd) and phosphate in the Southern Ocean are related to biology and hydrography. During a period of 18 days between transects 5/6 and 11, a phytoplankton spring bloom developed in the Polar Frontal region. Upper water Cd concentrations were not depleted and ranged from 0.2 to 0.8 nM at about 10 m depth. These relatively high Cd concentrations are attributed to upwelling of Upper Circumpolar Deep Water (0.5–1.2 nM in the core) in combination with low biological productivity (0.2 to 0.3 mg m⁻³ chlorophyll-a, 0.3 g C m⁻² d⁻¹). Total particulate Cd concentrations at 40 m depth were between 0.02 and 0.14 nM with the maximum in concentration in the Polar Frontal region. Most of the particulate Cd at this depth (85–94%) was detected in the first phase of a sequential chemical leaching treatment which includes adsorbed Cd as well as Cd incorporated in algae. The Polar Frontal region was characterized by minima in Cd concentration and Cd/phosphate ratio of seawater at both transects; values were the lowest at transect 11 after development of the spring bloom which was dominated by diatoms. This decreasing Cd/phosphate ratio in seawater during spring bloom development was attributed to preferential Cd gross uptake which more than compensated the process of preferential Cd recycling. Within the Upper Circumpolar Deep Water, Cd showed a maximum in concentration similar to that of the major nutrients. Both the Cd concentration and the Cd/phosphate ratio of the deeper water increased in southern direction, from 0.4 to 0.7 nM and from 0.2 to 0.3 nM/μM, respectively. Antarctic Intermediate Water has a Cd concentration of 0.21 nM with a Cd/phosphate ratio of 0.10 nM/μM. In Antarctic Bottom Water, Cd concentrations ranged from 0.60 to 0.82 nM.     

Determination of copper complexation with natural organic ligands in seawater by equilibration with MnO2 II. Experimental procedures and application to surface seawater

The MnO₂ adsorption method combined with voltammetry is proposed for the direct determination of metal complexation in seawater of various salinities as a more satisfactory alternative to direct voltammetric measurements and bioassay methods. A small quantity of MnO₂ is equilibrated with copper ions in filtered seawater. Natural organic ligands in the seawater compete for copper with the MnO₂. Total dissolved copper is measured by differential pulse anodic stripping voltammetry after filtration and acidification of the sample. Preconcentration of natural water samples is unnecessary and measurement is performed at the natural equilibrium pH of the aerated sample. The analytical limit of detection of the method depends on contamination from the filtration step, and for copper complexation a ligand concentration of 5 × 10^?8 M was obtained. The sensitivity can be increased by use of radioisotopes as tracers. The method is very versatile in that complexation of various metals may be determined by any analytical method that measures total dissolved metal concentrations. Neither organic ligands nor their complexes with copper adsorb on the MnO₂ at pH8, but at pH 1.8 MnO₂ is an efficient scavenger for electroactive organic material.Samples of surface water from the Irish Sea and the Atlantic Ocean were found to contain ligand concentrations of 1.7 × 10^?7 and 1.1 × 10^?7 M, with conditional stability constants (log values) of 9.84 ± 0.13 and 9.86 ± 0.23, respectively, at pH 8.0.     

Long lifetimes of β-glucosidase,leucine aminopeptidase,and phosphatase in Arctic seawater

The active lifetime of extracellular enzymes is a critical determinant of the effectiveness of enzyme production as a means for heterotrophic marine microbes to obtain organic substrates. Here, we report lifetimes of three classes of extracellular enzyme in Arctic seawater. We also investigated the relative importance of photochemical processes and particle-associated processes in inactivating extracellular enzymes. Enzyme inactivation in filtered seawater was slow, with apparent half-lives of enzyme activities on the order of hundreds of hours. The presence of particles (including cells) did not significantly change inactivation rates, suggesting that the long half-lives observed in filtered seawater were realistic for enzymes in unfiltered seawater. Phosphatase and leucine aminopeptidase were susceptible to photoinactivation, but only under high intensity UV-B and UV-C illumination; there was no evidence for increased inactivation rates under natural illumination at our study site in Ny Ålesund, Svalbard. Comparison of inactivation rates of commercially-obtained enzymes from non-marine sources with the extracellular enzymes naturally present in Arctic seawater suggests that the natural enzymes contain structural features that confer longer lifetimes, consistent with observations reported by others from a range of field sites that cell-free enzymes can contribute a substantial fraction of total hydrolytic activity in the water column.     

Ammonium input to the sea via large sewage outfalls—part 1: Tracing sewage in Southern California waters

Ammonium concentrations were found to be elevated near Southern California sewage outfalls; concentrations exceeding 3 μg-atom litre⁻¹ were measured 5 km from the discharge area. In stratified water, high values were found below 15 m, but in well-mixed water, high levels were detected at the surface. Subsurface high concentrations were associated with turbid layers, coliform bacteria and reduced oxygen levels. The distribution of ammonium correlated well with measured subsurface currents. The maximum concentration at the Whites Point outfall was 155 μg-atom litre⁻¹ at 27 m, about 2 km from the diffuser. Measurements of ammonium in sewage, compared with that in seawater at Whites Point, suggested that sewage was diluted up to 400-fold. Ammonium may be a useful tracer of the discharge of sewage in seawater.     

Calibration and evaluation of a carbonate microsensor for studies of the marine inorganic carbon system

Accurate and rapid determination of inorganic carbon constituents in ocean environments is important for understanding the carbon cycle, especially in the context of ocean-acidification research. A microsensor capable of directly measuring carbonate ion (CO₃ ^2–) concentrations would be desirable. In this study, a carbonate microsensor with a polymeric liquid membrane was fabricated, and two calibration methods were used to evaluate its performance. The first method was based on continuous titration. Small increments of HCl were added to seawater or Na₂CO₃ solution to adjust the total alkalinity and pH values and thus obtain a series of carbonate concentrations. The second method used a series of discrete standards. Varying amounts of HCl or NaOH were added to separate seawater aliquots, and the CO₃ ^2– concentration of each standard was calculated from the resulting total alkalinity and total dissolved inorganic carbon. Both methods were found to be adequate for achieving accurate calibration of the CO₃ ^2– sensor, and both are suitable for field work. The discrete standards method, however, is more convenient and may provide a better linear range at low CO₃ ^2– concentrations (detection range: 2–300 μmol/kg) than the continuous titration method in seawater (detection range: 10–250 μmol/kg). This CO₃ ^2– microsensor can be used for 5–7 d and detects changes in carbonate concentration as low as 2 μmol/kg in the inorganic carbon constituents of the environments where marine calcareous organisms grow. The CO₃ ^2– microelectrode was further assessed by applying it to the measurement of pore-water CO₃ ^2– concentration profiles in a marine sediment core.     

Using major nutrient concentrations to derive vertical movement of water masses in the coastal region of eastern Taiwan

Significant variations of phosphate and silicate concentrations have been observed in seawater from selected industrial pipelines at intake depths between 400 and 710 m off the Hualien coast, eastern Taiwan, revealing a strong vertical movement of water mass in this intermediate layer. An intensive monitoring experiment was carried out, in which pipeline seawater from three land-based pumping stations and seawater collected by a research vessel were obtained and analyzed in parallel during a 2-day observation period. The results showed clearly that the changes of nutrient concentrations in both pipeline and shipboard samples followed a semidiurnal cycle. The maximum vertical displacement occurred in the 300–800 m layer with a scale as surprisingly large as 100 m, when compared with that observed in other areas. The tidal cycles for different layers may not synchronize with surface tide or each other. Empirical equations have been implemented between nutrient concentrations and temperature for the Hualien off-shore area. The equation can be used to check temperature variation of the intermediate water by measuring either phosphate or silicate in pipeline seawater.     

Recovery of ammonium nitrogen by solvent extraction for the determination of relative 15N abundance in regeneration experiments

A new sample preparation method for ¹⁵N tracer measurement of ammonium regeneration in seawater is described. Ammonium nitrogen is incorporated into indophenol, extracted into dichloromethane and concentrated by evaporation. The isotopic ratio of the indophenol nitrogen is determined by emission spectroscopy. A small amount (0.2 μmol) of unlabeled nitrogen is introduced by reagents used during sample preparation. For samples of equal nitrogen content, the coefficient of variation is constant and the precision of the isotopic ratio determination is inversely related to the relative ¹⁵N abundance. Differences of 0.19 atom% ¹⁵N can be measured in 5.0 μM NH₄⁺ samples of 5.0 atom% ¹⁵N (SD = 0.07, n = 3, p = 0.05). Interface from forms of dissolved organic nitrogen prevalent in marine environments are negligible.     

海水悬浮物浓度原位测量技术进展

海水悬浮物指海水中的悬浮颗粒物和浮游微生物等，对海水悬浮物浓度进行准确的测量具有重要的意义。海水悬浮物浓度测量技术，无论是传统方法（即现场采水），还是现代方法（即间接测量方式，包括光学法和声学法等）都有一定的不足。本文主要综述了几种海水悬浮物浓度间接测量方法、国内外有关海水原位过滤测量的技术研发进展及深海原位微孔过滤技术。最后，对海水悬浮物原位过滤技术的发展方向进行了探讨。