A hybrid conductometric/spectrophotometric method for determining ionic strength of dilute aqueous solutions.

This work describes a novel conductometric/spectrophotometric method to determine the ionic strength (I) of dilute aqueous solutions (e.g., natural waters from rivers and lakes). Because I ≤ 0.01 mol kg-1 in such waters, precision as well as accuracy is of paramount importance. In current practice the ionic strength of natural waters is determined almost exclusively with conductometric measurements. We used solutions of artificial freshwater to assess the performance of two commonly used types of conductometric instruments and found that a conductivity probe systematically overestimated I while a salinometer systematically underestimated I. We therefore recommend here an empirical correction that can be easily implemented to improve the accuracy of both types of conductivity measurements. Additional improvement in measurements of I can be achieved by using that high-quality conductometric measurement as input to a hybrid conductometric/spectrophotometric procedure that makes use of robust quantitative characterizations of the influence of ionic strength on the dissociation characteristics of phosphate pH buffers and sulfonephthalein pH indicators. This approach mitigates systematic conductometric errors associated with solution composition, thus yielding measurements of ionic strength with substantially improved accuracy and precision. The method was validated by testing on a broad suite of artificial freshwaters (n = 64) with compositions that include the major ions present in dilute natural waters (Na+, K+, Mg2+, Ca2+, Cl-, HCO3-, and SO42-). This new hybrid method is applicable to waters of 0 ≤ I ≤ 0.01 mol kg-1 (i.e., electrical conductivity of up to 900 µS cm-1 at 25 °C), with an accuracy of ±0.0003 and a precision of ±0.0003.

Reef Metabolism Monitoring Methods and Potential Applications for Coral Restoration.

Coral reef metabolism measurements have been used by scientists for decades to track reef responses to the globe's changing carbon budget and project shifts in reef function. Here, we propose that metabolism measurement tools and methods could also be used to monitor reef ecosystem change in response to coral restoration. This review paper provides a general introduction to net ecosystem metabolism and carbon chemistry for coral reef ecosystems, followed by a review of five metabolism monitoring methods with potential for application to coral reef restoration monitoring. Selected methodologies included those with measurement scales appropriate to assess outplant arrays and whole reef ecosystem outcomes associated with restoration interventions. Subsequently we discuss how water column and CO2 chemistry could be used to address coral restoration monitoring research gaps and scale up from biological, colony-level metrics to ecosystem-scale function and performance assessments. Such function-based measurements could potentially be used to inform several goal-based monitoring objectives highlighted in the Coral Reef Restoration Monitoring Guide. Lastly, this review discusses important methodological factors, such as scale, reef type, and flow environment, that should be considered when determining which metabolism monitoring technique would be most appropriate for a reef restoration project.

Purification and Physical-Chemical Characterization of Bromocresol Purple for Carbon System Measurements in Freshwaters, Estuaries, and Oceans.

This work provides an algorithm to describe the salinity (S P) and temperature (T) dependence of the equilibrium and molar absorptivity characteristics of purified bromocresol purple (BCP, a pH indicator) over a river-to-sea range of salinity (0 ≤ S P ≤ 40). Based on the data obtained in this study, the pH of water samples can be calculated on the seawater pH scale as follows: pHSW = -log(K 2 e 2) + log((R - e 1)/(1 - Re 4)) where -log(K 2 e 2) = 4.981 - 0.1710S P 0.5 + 0.09428S P + 0.3794S P 1.5 + 0.0009129S P 2 + 310.2/T - 17.33S 1.5/T - 0.05895S P 1.5 ln T - 0.0005730S P 0.5 T, e 1 = 0.00049 ± 0.00029, and e 4 = -7.101 × 10-3 + 7.674 × 10-5 T + 1.361 × 10-5 S P. The term pHSW is the negative log of the hydrogen ion concentration determined on the seawater pH scale; R is the ratio of BCP absorbances (A) at 432 and 589 nm; K 2 is the equilibrium constant for the second BCP dissociation step; and e 1, e 2, and e 4 are BCP molar absorptivity ratios. A log(K 2 e 2) equation is also presented on the total pH scale. The e 4 value determined for purified BCP in this study can be used with previously published procedures to correct BCP absorbance measurements obtained using off-the-shelf (unpurified) BCP. This work provides a method for purifying BCP, fills a critical gap in the suite of available purified sulfonephthalein indicators, enables high-quality spectrophotometric measurements of total alkalinity, and facilitates pH measurements in freshwater, estuarine, and ocean environments within the range 4.0 ≤ pH ≤ 7.5.

Purification and characterization of thymol blue for spectrophotometric pH measurements in rivers, estuaries, and oceans.

Thymol blue (TB) is one of a suite of indicator dyes appropriate for spectrophotometric determinations of the pH of aqueous solutions. For measurements of seawater pH, meta-cresol purple (mCP) is most often used, but TB is especially well suited for measurements in surface or shallow waters where the pH may exceed the optimal indicating range of mCP (e.g., due to photosynthesis). This work presents flash chromatography procedures for purifying commercially available TB and describes physical-chemical characteristics of the purified dye, thus enabling the acquisition of spectrophotometric pH measurements over a wide range of practical salinities (SP) and temperatures (T). The essential TB characteristics for 0 ≤ SP ≤ 40 and 278.15 ≤ T ≤ 308.15 K are described by: pHT=-TB(log(K2Te2))+log((R-e1)/(1-Re4)) TB(logK2Te2)=6.6942-0.001129Sp0.5T-0.5926T-0.5Sp+619.40/T+0.1441SP-0.02591Sp1.5+0.0034Sp2-0.0001754Sp2.5 e1 = -0.00132 + 0.00001600T e4 = -0.005042 + 0.0002094T + 0.01916SP0.5/T where pHT is pH determined on the total hydrogen ion concentration scale; R is the ratio of TB absorbances (A) at 435 and 596 nm (596A/435A); K2T is the equilibrium constant for the second TB dissociation step on the total scale; and e1, e2, and e4 are TB molar absorptivity ratios. This characterization was developed in a manner that ensures consistency with the primary TRIS buffer standards used in previously published characterizations of mCP. With this characterization, TB joins mCP as a sulfonephthalein indicator that has been characterized over the ranges of salinity and temperature required to make high-quality pH measurements in rivers, estuaries, and the open ocean. The full characterization of purified TB reported here extends the upper range of pHT that can be accessed with precise spectrophotometric measurements by approximately 0.50 pH units.

Spectrophotometric determination of pH and carbonate ion concentrations in seawater: Choices, constraints and consequences.

Accurate and precise marine CO2 system measurements are important for marine carbon cycle research and investigations of ocean acidification. Seawater pH is important because it can be used to characterize a wide range of chemical and biogeochemical processes. Saturation states of calcium carbonate minerals, which are directly proportional to carbonate ion concentration ([CO32-]), influence biogenic calcification and rates of carbonate dissolution. Spectrophotometric pH and carbonate ion measurements can both benefit greatly from the high sensitivity, stability, consistency and processing speed made possible through automation. Spectrophotometric methods are well-suited for shipboard, underway and in situ deployments under harsh conditions. Spectrophotometric pH measurements typically have a reproducibility of 0.0004-0.001 for shipboard and laboratory measurements and 0.0014-0.004 for in situ measurements. Shipboard spectrophotometric measurements of [CO32-] are becoming common on research expeditions. This review highlights the development of methods and instrumentation for spectrophotometric pH and [CO32-] measurements, and discusses the pros and cons of current technology. A comprehensive summary of the analytical merits of different flow analysis instruments is given. Aspects of measurement protocols that bear on the quality of pH and [CO32-] measurements, such as indicator purification, sample pretreatment, etc., are also described. Based on three decades of experience with seawater analysis, this review includes method recommendations and perspectives directly applicable or potentially applicable to pH and [CO32-] analysis of seawater.

Avoiding timescale bias in assessments of coastal wetland vertical change.

There is concern that accelerating sea-level rise will exceed the vertical growth capacity of coastal-wetland substrates in many regions by the end of this century. Vertical vulnerability estimates rely on measurements of accretion and/or surface-elevation-change derived from soil cores and/or surface elevation tables (SETs). To date there has not been a broad examination of whether the multiple timescales represented by the processes of accretion and elevation change are equally well-suited for quantifying the trajectories of wetland vertical change in coming decades and centuries. To examine the potential for timescale bias in assessments of vertical change, we compared rates of accretion and surface elevation change using data derived from a review of the literature. In the first approach, average rates of elevation change were compared with timescale-averaged accretion rates from six regions around the world where sub-decadal, decadal, centennial, and millennial timescales were represented. Second, to isolate spatial variability, temporal comparisons were made for regionally unique environmental categories within each region. Last, comparisons were made of records from sites where SET-MH stations and radiometric measurements were co-located in close proximity. We find that rates vary significantly as a function of measurement timescale and that the pattern and magnitude of variation between timescales are location-specific. Failure to identify and account for temporal variability in rates will produce biased assessments of the vertical change capacity of coastal wetlands. Robust vulnerability assessments should combine accretion rates from multiple timescales with the longest available SET record to provide long-term context for ongoing monitoring observations and projections.

Spectrophotometric calibration procedures to enable calibration-free measurements of seawater calcium carbonate saturation states.

A simple protocol was developed to measure seawater calcium carbonate saturation states (Ωspec) spectrophotometrically. Saturation states are typically derived from the separate measurement of two other carbon system parameters, with each requiring unique instrumentation and often complex measurement protocols. Using the new protocol, the only required equipment is a thermostatted laboratory spectrophotometer. For each seawater sample, spectrophotometric measurements of pH (visible absorbance) are made in paired optical cells, one with and one without added nitric acid. Ultraviolet absorbance is measured to determine the amount of added acid based on the direct proportionality between nitrate concentration and UV absorbance. Coupled measurements of pH and the alkalinity change that accompanies the nitric acid addition allow calculation of a seawater sample's original carbonate ion concentration and saturation state. These paired absorbance measurements yield Ωspec (and other carbonate system parameters), with each sample requiring about 12 min processing time. Initially, an instrument-specific nitrate molar absorptivity coefficient must be determined (due to small but significant discrepancies in instrumental wavelength calibrations), but thereafter no further calibration is needed. In this work, the 1σ precision of replicate measurements of aragonite saturation state was found to be 0.020, and the average difference between Ωspec and Ω calculated conventionally from measured total alkalinity and pH (Ωcalc) was -0.11% ±â€¯0.96% (a level of accuracy comparable to that obtained from spectrophotometric measurements of carbonate ion concentration). Over the entire range of experimental conditions, 0.97 < Ω < 3.17 (n = 125), all measurements attained the Global Ocean Acidification Observing Network's "weather level" goal for accuracy and 90% attained the more stringent "climate level" goal. When Ωspec was calculated from averages of duplicate samples (n = 56), the precision improved to 0.014 and the average difference between Ωspec and Ωcalc improved to -0.11% ±â€¯0.73%. Additionally, 97% of the duplicate-based Ωspec measurements attained the "climate level" accuracy goal. These results indicate that the simple measurement protocol developed in this work should be widely applicable for monitoring fundamental seawater changes associated with ocean acidification.

Spectrophotometric Determination of Carbonate Ion Concentrations: Elimination of Instrument-Dependent Offsets and Calculation of In Situ Saturation States.

This work describes an improved algorithm for spectrophotometric determinations of seawater carbonate ion concentrations ([CO32-]spec) derived from observations of ultraviolet absorbance spectra in lead-enriched seawater. Quality-control assessments of [CO32-]spec data obtained on two NOAA research cruises (2012 and 2016) revealed a substantial intercruise difference in average Δ[CO32-] (the difference between a sample's [CO32-]spec value and the corresponding [CO32-] value calculated from paired measurements of pH and dissolved inorganic carbon). Follow-up investigation determined that this discordance was due to the use of two different spectrophotometers, even though both had been properly calibrated. Here we present an essential methodological refinement to correct [CO32-]spec absorbance data for small but significant instrumental differences. After applying the correction (which, notably, is not necessary for pH determinations from sulfonephthalein dye absorbances) to the shipboard absorbance data, we fit the combined-cruise data set to produce empirically updated parameters for use in processing future (and historical) [CO32-]spec absorbance measurements. With the new procedure, the average Δ[CO32-] offset between the two aforementioned cruises was reduced from 3.7 µmol kg-1 to 0.7 µmol kg-1, which is well within the standard deviation of the measurements (1.9 µmol kg-1). We also introduce an empirical model to calculate in situ carbonate ion concentrations from [CO32-]spec. We demonstrate that these in situ values can be used to determine calcium carbonate saturation states that are in good agreement with those determined by more laborious and expensive conventional methods.

Measuring ocean acidification: new technology for a new era of ocean chemistry.

Human additions of carbon dioxide to the atmosphere are creating a cascade of chemical consequences that will eventually extend to the bottom of all the world's oceans. Among the best-documented seawater effects are a worldwide increase in open-ocean acidity and large-scale declines in calcium carbonate saturation states. The susceptibility of some young, fast-growing calcareous organisms to adverse impacts highlights the potential for biological and economic consequences. Many important aspects of seawater CO2 chemistry can be only indirectly observed at present, and important but difficult-to-observe changes can include shifts in the speciation and possibly bioavailability of some life-essential elements. Innovation and invention are urgently needed to develop the in situ instrumentation required to document this era of rapid ocean evolution.

Flow injection analysis of trace chromium (VI) in drinking water with a liquid waveguide capillary cell and spectrophotometric detection.

Hexavalent chromium (Cr(VI)) is an acknowledged hazardous material in drinking waters. As such, effective monitoring and assessment of the risks posed by Cr(VI) are important analytical objectives for both human health and environmental science. However, because of the lack of highly sensitive, rapid, and simple procedures, a relatively limited number of studies have been carried out in this field. Here we report a simple and sensitive analytical procedure of flow injection analysis (FIA) for sub-nanomolar Cr(VI) in drinking water samples with a liquid core waveguide capillary cell (LWCC). The procedure is based on a highly selective reaction between 1, 5-diphenylcarbazide and Cr(VI) under acidic conditions. The optimized experimental parameters included reagent concentrations, injection volume, length of mixing coil, and flow rate. Measurements at 540 nm, and a 650-nm reference wavelength, produced a 0.12-nM detection limit. Relative standard deviations for 1, 2, and 10 nM samples were 5.6, 3.6, and 0.72 % (n = 9), and the analysis time was <2 min sample(-1). The effects of salinity and interfering ions, especially Fe(III), were evaluated. Using the FIA-LWCC method, different sources of bottled waters and tap waters were examined. The Cr(VI) concentrations of the bottled waters ranged from the detection limit to ∼20 nM, and tap waters collected from the same community supply had Cr(VI) concentration around 14 nM.

In situ spectrophotometric measurement of dissolved inorganic carbon in seawater.

Autonomous in situ sensors are needed to document the effects of today's rapid ocean uptake of atmospheric carbon dioxide (e.g., ocean acidification). General environmental conditions (e.g., biofouling, turbidity) and carbon-specific conditions (e.g., wide diel variations) present significant challenges to acquiring long-term measurements of dissolved inorganic carbon (DIC) with satisfactory accuracy and resolution. SEAS-DIC is a new in situ instrument designed to provide calibrated, high-frequency, long-term measurements of DIC in marine and fresh waters. Sample water is first acidified to convert all DIC to carbon dioxide (CO2). The sample and a known reagent solution are then equilibrated across a gas-permeable membrane. Spectrophotometric measurement of reagent pH can thereby determine the sample DIC over a wide dynamic range, with inherent calibration provided by the pH indicator's molecular characteristics. Field trials indicate that SEAS-DIC performs well in biofouling and turbid waters, with a DIC accuracy and precision of ∼2 µmol kg(-1) and a measurement rate of approximately once per minute. The acidic reagent protects the sensor cell from biofouling, and the gas-permeable membrane excludes particulates from the optical path. This instrument, the first spectrophotometric system capable of automated in situ DIC measurements, positions DIC to become a key parameter for in situ CO2-system characterizations.

Baseline monitoring of the western Arctic Ocean estimates 20% of Canadian basin surface waters are undersaturated with respect to aragonite.

Marine surface waters are being acidified due to uptake of anthropogenic carbon dioxide, resulting in surface ocean areas of undersaturation with respect to carbonate minerals, including aragonite. In the Arctic Ocean, acidification is expected to occur at an accelerated rate with respect to the global oceans, but a paucity of baseline data has limited our understanding of the extent of Arctic undersaturation and of regional variations in rates and causes. The lack of data has also hindered refinement of models aimed at projecting future trends of ocean acidification. Here, based on more than 34,000 data records collected in 2010 and 2011, we establish a baseline of inorganic carbon data (pH, total alkalinity, dissolved inorganic carbon, partial pressure of carbon dioxide, and aragonite saturation index) for the western Arctic Ocean. This data set documents aragonite undersaturation in ≈ 20% of the surface waters of the combined Canada and Makarov basins, an area characterized by recent acceleration of sea ice loss. Conservative tracer studies using stable oxygen isotopic data from 307 sites show that while the entire surface of this area receives abundant freshwater from meteoric sources, freshwater from sea ice melt is most closely linked to the areas of carbonate mineral undersaturation. These data link the Arctic Ocean's largest area of aragonite undersaturation to sea ice melt and atmospheric CO2 absorption in areas of low buffering capacity. Some relatively supersaturated areas can be linked to localized biological activity. Collectively, these observations can be used to project trends of ocean acidification in higher latitude marine surface waters where inorganic carbon chemistry is largely influenced by sea ice meltwater.

Evaluation of reagentless pH modification for in situ ocean analysis: determination of dissolved inorganic carbon using mass spectrometry.

RATIONALE: In situ analytical techniques that require the storage and delivery of reagents (e.g., acidic or basic solutions) have inherent durability limitations. The reagentless electrolytic technique for pH modification presented here was developed primarily to ease and to extend the longevity of dissolved inorganic carbon (DIC) determinations in seawater, but can also be used for other analytical methods. DIC, a primary carbon dioxide (CO(2)) system variable along with alkalinity, controls seawater pH, carbonate saturation state, and CO(2) fugacity. Determinations of these parameters are central to an understanding of ocean acidification and global climate change. METHODS: Electrodes fabricated with electroactive materials, including manganese(III) oxide (Mn(2)O(3)) and palladium (Pd), were examined for potential use in electrolytic acidification. In-line acidification techniques were evaluated using a bench-top membrane introduction mass spectrometry (MIMS) setup to determine the DIC content of artificial seawater. Linear least-squares (LLSQ) calibrations for DIC concentration determinations over a range between 1650 and 2400 µmol kg(-1) were obtained, using both the novel electrolytic and conventional acid addition techniques. RESULTS: At sample rates of 4.5 mL min(-1), electrodes clad with Mn(2)O(3) and Pd were able to change seawater pH from 7.6 to 2.8 with a power consumption of less than 3 W. Although calibration curves were influenced by sampling rates at a flow of 4.5 mL min(-1), the 1σ measurement precision for DIC was of the order of ±20 µmol kg(-1). CONCLUSIONS: Calibrations obtained with the novel reagentless technique and the in-line addition of strong acid showed similar capabilities for DIC quantification. However, calculations of power savings for the reagentless technique relative to the mechanical delivery of stored acid demonstrated substantial advantages of the electrolytic technique for long-term deployments (>1 year).

Spectrophotometric measurement of calcium carbonate saturation states in seawater.

Measurements of ocean pH and carbonate ion concentrations in the North Pacific and Arctic Oceans were used to determine calcium carbonate saturation states (Ω(CaCO(3))) from spectrophotometric methods alone. Total carbonate ion concentrations, [CO(3)(2-)](T), were for the first time at sea directly measured using Pb(II) UV absorbance spectra. The basis of the method is given by the following: [formula see text] where (CO(3))ß(1) is the PbCO(3)(0) formation constant, e(i) are molar absorptivity ratios, and R = (250)A/(234)A (ratio of absorbances measured at 250 and 234 nm). On the basis of shipboard and laboratory Pb(II) data and complementary carbon-system measurements, the experimental parameters were determined to be (25 °C) the following: [formula see text]. The resulting mean difference between the shipboard spectrophotometric and conventional determinations of [CO(3)(2-)](T) was ±2.03 µmol kg(-1). The shipboard analytical precision of the Pb(II) method was ∼1.71 µmol kg(-1) (2.28%). Spectrophotometric [CO(3)(2-)](T) and pH(T) were then combined to calculate Ω(CaCO(3)). For the case of aragonite, 95% of the spectrophotometric aragonite saturation states (Ω(Aspec)) were within ±0.06 of the conventionally calculated values (Ω(Acalc)) when 0.5 ≤ Ω(A) ≤ 2.0. When Ω(A) > 2.0, 95% of the Ω(Aspec) values were within ±0.18 of Ω(Acalc). Our shipboard experience indicates that spectrophotometric determinations of [CO(3)(2-)](T) and Ω(CaCO(3)) are straightforward, fast, and precise. The method yields high-quality measurements of two important, rapidly changing aspects of ocean chemistry and offers capabilities suitable for long-term automated in situ monitoring.

Determination of nanomolar chromate in drinking water with solid phase extraction and a portable spectrophotometer.

Determination of chromate at low concentration levels in drinking water is an important analytical objective for both human health and environmental science. Here we report the use of solid phase extraction (SPE) in combination with a custom-made portable light-emitting diode (LED) spectrophotometer to achieve detection of chromate in the field at nanomolar levels. The measurement chemistry is based on a highly selective reaction between 1,5-diphenylcarbazide (DPC) and chromate under acidic conditions. The Cr-DPC complex formed in the reaction can be extracted on a commercial C18 SPE cartridge. Concentrated Cr-DPC is subsequently eluted with methanol and detected by spectrophotometry. Optimization of analytical conditions involved investigation of reagent compositions and concentrations, eluent type, flow rate (sample loading), sample volume, and stability of the SPE cartridge. Under optimized conditions, detection limits are on the order of 3 nM. Only 50 mL of sample is required for an analysis, and total analysis time is around 10 min. The targeted analytical range of 0-500 nM can be easily extended by changing the sample volume. Compared to previous SPE-based spectrophotometric methods, this analytical procedure offers the benefits of improved sensitivity, reduced sample consumption, shorter analysis time, greater operational convenience, and lower cost.

Spectrophotometric calibration of pH electrodes in seawater using purified m-cresol purple.

This work examines the use of purified meta-cresol purple (mCP) for direct spectrophotometric calibration of glass pH electrodes in seawater. The procedures used in this investigation allow for simple, inexpensive electrode calibrations over salinities of 20-40 and temperatures of 278.15-308.15 K without preparation of synthetic Tris seawater buffers. The optimal pH range is ∼7.0-8.1. Spectrophotometric calibrations enable straightforward, quantitative distinctions between Nernstian and non-Nernstian electrode behavior. For the electrodes examined in this study, both types of behavior were observed. Furthermore, calibrations performed in natural seawater allow direct determination of the influence of salinity on electrode performance. The procedures developed in this study account for salinity-induced variations in liquid junction potentials that, if not taken into account, would create pH inconsistencies of 0.028 over a 10-unit change in salinity. Spectrophotometric calibration can also be used to expeditiously determine the intercept potential (i.e., the potential corresponding to pH 0) of an electrode that has reliably demonstrated Nernstian behavior. Titrations to ascertain Nernstian behavior and salinity effects can be undertaken relatively infrequently (∼weekly to monthly). One-point determinations of intercept potential should be undertaken frequently (∼daily) to monitor for stable electrode behavior and ensure accurate potentiometric pH determinations.

Flow injection analysis of nanomolar silicate using long pathlength absorbance spectroscopy.

Determination of silicate at low concentrations (i.e., nanomolar levels) is an important analytical objective for both marine science and the semiconductor industry. Here we report the use of flow injection analysis (FIA) in combination with long pathlength liquid core waveguide (LCW) spectrometry to achieve detection limits for dissolved silica on the order of 10nM. Sample throughput for the simple, automated analytical apparatus used in this work is 12h(-1) at low levels of dissolved silica; this rate can be increased by a factor of three for higher (micromolar) levels of dissolved silica. The analytical protocol is based on the reaction of silicate with ammonium molybdate to form a yellow silicomolybdate complex, which is subsequently reduced to silicomolybdenum blue by ascorbic acid. Optimization of the FIA procedure included consideration of the compositions and concentrations of reagents, volume of the injection loop, flow rate conditions, and lengths of mixing coils. The interference by phosphate was examined and eliminated through addition of oxalic acid. The dissolved silica detection limit of 7.2nM in pure water is consistent with the strictest standard for the semiconductor industry, and the 9.0nM detection limit for seawater shows that this analytical method is also suitable for oligotrophic ocean waters. The targeted analytical range of 10nM to 5µM can be easily extended to higher concentrations without altering the experimental hardware-i.e., by simply changing flow rates or selecting alternative analytical wavelengths. Compared to previously published LCW-based spectrophotometric methods, this analytical system exhibits improved sensitivity, reduced sample consumption, and higher sample throughput.

Purification and characterization of meta-cresol purple for spectrophotometric seawater pH measurements.

Spectrophotometric procedures allow rapid and precise measurements of the pH of natural waters. However, impurities in the acid-base indicators used in these analyses can significantly affect measurement accuracy. This work describes HPLC procedures for purifying one such indicator, meta-cresol purple (mCP), and reports mCP physical-chemical characteristics (thermodynamic equilibrium constants and visible-light absorbances) over a range of temperature (T) and salinity (S). Using pure mCP, seawater pH on the total hydrogen ion concentration scale (pHT) can be expressed in terms of measured mCP absorbance ratios (R = λ2A/(λ1)A) as follows: [formula in text] where -log(K(2)Te2) = a + (b/T) + c ln T ­ dT; a = -246.64209 + 0.315971S + 2.8855 × 10(-4)S2; b = 7229.23864 ­ 7.098137S ­ 0.057034S2; c = 44.493382 ­ 0.052711S; d = 0.0781344; and mCP molar absorbance ratios (ei) are expressed as e1 = -0.007762 + 4.5174 × 10(-5)T and e3/e2 = -0.020813 + 2.60262 × 10(-4)T + 1.0436 × 10(-4) (S ­ 35). The mCP absorbances, λ1A and λ2A, used to calculate R are measured at wavelengths (λ) of 434 and 578 nm. This characterization is appropriate for 278.15 ≤ T ≤ 308.15 and 20 ≤ S ≤ 40.

Phytoplankton carbon fixation gene (RuBisCO) transcripts and air-sea CO(2) flux in the Mississippi River plume.

River plumes deliver large quantities of nutrients to oligotrophic oceans, often resulting in significant CO(2) drawdown. To determine the relationship between expression of the major gene in carbon fixation (large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBisCO) and CO(2) dynamics, we evaluated rbcL mRNA abundance using novel quantitative PCR assays, phytoplankton cell analyses, photophysiological parameters, and pCO(2) in and around the Mississippi River plume (MRP) in the Gulf of Mexico. Lower salinity (30-32) stations were dominated by rbcL mRNA concentrations from heterokonts, such as diatoms and pelagophytes, which were at least an order of magnitude greater than haptophytes, alpha-Synechococcus or high-light Prochlorococcus. However, rbcL transcript abundances were similar among these groups at oligotrophic stations (salinity 34-36). Diatom cell counts and heterokont rbcL RNA showed a strong negative correlation to seawater pCO(2). While Prochlorococcus cells did not exhibit a large difference between low and high pCO(2) water, Prochlorococcus rbcL RNA concentrations had a strong positive correlation to pCO(2), suggesting a very low level of RuBisCO RNA transcription among Prochlorococcus in the plume waters, possibly due to their relatively poor carbon concentrating mechanisms (CCMs). These results provide molecular evidence that diatom/pelagophyte productivity is largely responsible for the large CO(2) drawdown occurring in the MRP, based on the co-occurrence of elevated RuBisCO gene transcript concentrations from this group and reduced seawater pCO(2) levels. This may partly be due to efficient CCMs that enable heterokont eukaryotes such as diatoms to continue fixing CO(2) in the face of strong CO(2) drawdown. Our work represents the first attempt to relate in situ microbial gene expression to contemporaneous CO(2) flux measurements in the ocean.