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Marine Chemistry 109 (2008) 18 28 www.elsevier.com/locate/marchem

A sensor for in situ indicator-based measurements of seawater pH

Matthew P. Seidel a,1 , Michael D. DeGrandpre a,, Andrew G. Dickson b
b a Department of Chemistry, The University of Montana, 32 Campus Drive, Missoula, Montana 59812, United States Scripps Institution of Oceanography, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, United States

Received 12 June 2007; received in revised form 6 November 2007; accepted 28 November 2007 Available online 15 December 2007

Abstract Indicator-based spectrophotometric pH methods are now proven and commonly used for analysis of ocean samples; however, no autonomous system for long-term in situ applications has been developed based on this method. We describe herein an autonomous indicator-based pH sensor for seawater applications adapted from a design originally developed for freshwater pH measurements (SAMI-pH). The new SAMI-pH uses a different pH indicator, flow cell design, detection system, and mixing configuration to improve upon the freshwater performance. A new method was also tested that utilizes an indicator concentration gradient in the sample to correct for the pH perturbation caused by the indicator. With these design changes, laboratory tests found the precision improved from 0.004 to 0.0007 and the accuracy improved from 0.0030 to + 0.0017 based on comparisons with benchtop UV/Vis measurements. In situ testing of two SAMI-pH instruments was completed off the pier at Scripps Institution of Oceanography. The average pH offset between the two instruments over the 22 d deployment period was 0.0042 0.0126 (n = 883), with the precision primarily regulated by large spatial and temporal variability at the site. The results demonstrate that the SAMIpH can provide drift-free and precise pH measurements in adverse measurement conditions (extensive fouling and large tidal variability). With the current battery power (18 alkaline D-cells), the system can be deployed for periods up to 2 months with a 0.5 h measurement frequency. 2008 Elsevier B.V. All rights reserved.
Keywords: pH sensor; in situ; autonomous; spectrophotometry; inorganic carbon; marine chemistry

1. Introduction Quantifying and understanding the impacts of oceanic uptake of anthropogenic CO2 is an important contemporary problem that has brought intense focus on the ocean inorganic carbon system. The rapid rise in atmospheric CO2 has increased the concentration of CO2 in the oceans
Corresponding author. Tel.: +1 406 243 4118; fax: +1 406 243 4227. E-mail address: michael.degrandpre@umontana.edu (M.D. DeGrandpre). 1 Tel.: +1 406 243 4118; fax: +1 406 243 4227. 0304-4203/$ - see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.marchem.2007.11.013

(Peng et al., 2003). Surface water pH has already decreased by 0.1 units and the continued uptake of CO2 is expected to decrease surface ocean pH by 0.20.3 units over the next century (Orr et al., 2005). A decrease in pH of this magnitude could have large-scale impacts on marine ecosystems (Orr et al., 2005), will result in decreased uptake of CO2 by the ocean (Sabine et al., 2004), and will affect burial rates of organic and inorganic carbon (Feely et al., 2004). Understanding the inorganic carbon cycle is also key to understanding and predicting net community metabolism, airsea gas exchange, and export of carbon from the surface to depth. New and

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improved autonomous techniques are needed that can collect high quality, high temporal resolution CO2 data for characterizing both long and short term changes in the marine inorganic carbon system. Seawater pH has become a more appealing inorganic carbon parameter because improvements in the spectrophotometric method have made it consistent with the other CO2 system parameters (e.g., Byrne and Breland, 1989) and it is readily measurable on ships with commercially available instrumentation. From our standpoint, another advantage is that spectrophotometric pH methods are, ostensibly, more easily adapted for in situ measurements. If pH is used in inorganic carbon calculations, it is highly desirable that the calculated dissolved inorganic carbon (DIC), total alkalinity (AT) or pCO2 be comparable in precision and accuracy to measured values, i.e., 1.5 mol kg 1 (Byrne et al., 1999), 2.0 mol kg 1 (Dickson et al., 2003), and 1 atm (Murphy et al., 1998), respectively. To illustrate the sensitivity of calculated DIC and pCO2 to pH uncertainty, we used the CO2 equilibrium model CO2SYS (Lewis and Wallace, 1998). The carbonic acid constants in Mehrbach et al. (1973) refit by Dickson and Millero (1987) were used because these constants give the most accurate calculations of inorganic carbon parameters from spectrophotometric pH measurements (Clayton et al., 1995; Byrne et al., 1999). Calculations were made using a constant alkalinity of 2250 mol kg 1 over the pH range 7.78.3. For a pH uncertainty of 0.005, the error in DIC ranges from 2.03.5 mol kg 1, lowest at the lower pHs typical of deep water. If the AT precision stated above is included in the calculation, the pH accuracy needs to be 0.002 to estimate DIC within 1.5 mol kg 1. This level of uncertainty is adequate for most applications. Similar calculations for pCO2 reveal that the pH error is more critical at lower pH values where pCO2 is higher. A pH uncertainty of 0.002 gives a pCO2 error of 5.0 atm at pH 7.7 (pCO2 = 980 atm) and an error of 1.1 atm at pH 8.3 (pCO2 = 182atm). Previous shipboard studies have obtained pH accuracy (which typically limits the pH uncertainty) from 0.0010.005 and precision from 0.0010.0004 (Byrne and Breland, 1989; Clayton et al., 1995; Tapp et al., 2000; Bellerby et al., 2002). Some work on in situ spectrophotometric pH sensors has been undertaken. An instrument for obtaining vertical depth profiles of pH and pCO2 from a ship has been developed (Nakano et al., 2006). The instrument collected data to 1000 m with accuracy of 0.002 0.005 pH units when compared to discrete samples. The SEAS pH instrument, designed for in situ spectrophotometric pH and other inorganic carbon measurements, was tested in seawater and freshwater (Liu et al., 2006). Shipboard

depth profiles in the Equatorial Pacific and Gulf of Mexico were collected. The in situ precision was 0.0014 while accuracy was not reported. Both the Nakano et al. (2006) and Liu et al. (2006) designs require high power (67 W) light sources, which limit the duration they can be deployed in the field. We have also previously presented the design and performance of an in situ spectrophotometric pH system, optimized for freshwater measurements (Martz et al., 2003). The Submersible Autonomous Moored Instrument for pH, or SAMI-pH, accuracy and precision was 0.003 and 0.004 pH units (n = 16), respectively, when deployed in a river over an 8 d period. No in situ indicator-based pH sensors, however, have been deployed for extended periods (weeks to months) in a marine environment. In this paper, we describe the optimization of the Martz et al. (2003) design for seawater applications. The new SAMIpH uses a different pH indicator, flow cell design, detection system, and mixing configuration to improve upon the freshwater performance. A 22 d in situ seawater pH time-series is presented and demonstrates that the SAMI-pH sensor is capable of collecting accurate and precise long-term in situ data on ocean moorings and other unmanned platforms. 2. Principle of operation A theoretical basis for the spectrophotometric determination of pH is given in Clayton and Byrne (1993) and readers should refer to that article and other articles by Byrne et al. for a more in depth discussion. The method is based upon the equilibrium reaction of a pH indicator, usually a diprotic sulfonephthalein indicator, HL H L2
pK a

where HL and L2 are the protonated and deprotonated forms of the indicator and pKa is the second apparent dissociation constant. The acidic H2L form is not present at seawater pHs. The sulfonephthalein indicator m-cresol purple (mCP) was selected because it has been extensively used in past studies (Clayton and Byrne, 1993; Clayton et al., 1995; Byrne et al., 1999; DOE, 1994). Combining the log form of the indicator equilibrium expression and Beer's Law yields:   R e1 pH pK 2 a log e2 Re3 where R is the absorbance ratio A2/A1 corresponding to the peak absorbance wavelengths 2 and 1 of the L2 and HL forms, respectively, and the eis are the temperature-

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M.P. Seidel et al. / Marine Chemistry 109 (2008) 1828

dependent ratios of the molar absorptivities () of the two species at both 1 and 2, e1
HL e578 HL e434

e2

HL e434

L e578

e3

HL e434

L e434

For mCP, the s are measured at the peak absorbance wavelengths for L2 (578 nm) and HL (434 nm). The adaptation of this methodology for autonomous in situ applications requires special considerations to retain the accuracy and precision of the benchtop method. Most lab measurements are performed at a fixed temperature, typically 25 C (DOE, 1994). For in situ applications, temperature is not controlled and the temperature dependence of pKa and eis must be accurately known. The eis also depend upon the detection system optical bandpass and must be determined for that bandpass and literature values cannot be used. In situ instruments, in particular, may use a larger bandpass to increase light throughput to reduce the light source power requirements. Moreover, when using a short pathlength optical cell (for reasons described below), the pH indicator solution can significantly alter the sample pH thus requiring some method to correct for the pH perturbation. Most benchtop methods use 10 cm pathlength cuvets with 5 10 6 mol kg 1 indicator (e.g., Clayton and Byrne, 1993). At this concentration, the magnitude of the perturbation is typically b 0.001 pH units (Chierici et al., 1999). Even so, various methods have been used to account for the pH perturbation (Zhang and Byrne, 1996; Bellerby et al., 2002). In the design presented here, we use a custom-

made 1.1 cm pathlength flow cell to maximize light throughput and minimize the light source power requirements and flush volume. Light throughput with a 0.5 W tungsten lamp gives sufficient signal-to-noise to achieve the desired level of absorbance precision. From our observations, throughput drops off roughly proportionally to pathlength, so a 1 W lamp is required to achieve the same signal-to-noise for a 2 cm pathlength cell. The short pathlength also reduces the pumping required to flush the system completely for blank measurements. These power savings allow for longer deployments, critical for an autonomous system. However, because the 1.1 cm cell requires indicator concentrations 10 times greater than 10 cm cells, the pH perturbation becomes significant. To address this problem, a perturbationcorrection methodology suggested by Martz et al. (2003) has been thoroughly evaluated here, as described below.
3. Methods 3.1. Instrument modifications Two instruments, SAMI-47 and SAMI-51, with 47 and 51 corresponding to their serial numbers, were manufactured by Sunburst Sensors, LLC with modifications to the Martz et al. (2003) design. The new design uses mCP instead of cresol red indicator because the mCP response better matches the full pH range of seawater. A commercially available static mixer (described below) replaced a coiled tube mixer to more effectively mix the indicator and sample. A custom made z-configuration flow cell (Fig. 1) replaced the 90o flow path optical cell used in the original SAMI design (DeGrandpre et al., 1995; Martz et al.,

Fig. 1. Instrument design for the seawater SAMI-pH modified from the SAMI-pH for freshwater (Martz et al., 2003). NO and NC refer to the normally open and closed valve settings. Dotted lines represent fiber optics.

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2003) to reduce bubble trapping that occasionally occurred in the 90 flow cell. Because of the orientation of the inlet relative to the optical channel, bubbles are more easily swept through the zconfiguration. To improve light throughput, the spectrograph detection system was replaced with a fiber optic splitter connected to three silicon photodiode detectors with discrete interference filters (Yuan and DeGrandpre, 2006). 3.2. Flow design The sample and indicator solutions are connected to the normally open and normally closed ports, respectively, of a three-way solenoid valve (161K031, Neptune Research Inc.) (Fig. 1). A low power 50 l per pulse solenoid pump (LPLA1210050L, The Lee Co.) draws sample or indicator through the valve and dispenses it into the downstream plumbing. The pump and valve are enclosed in a silicone oilfilled housing with a diaphragm for pressure equilibration. The indicator solution, described below, is contained in a 150 ml gasimpermeable bag (Pollution Measurement Corp.). The static mixer, developed for HPLC, is a 350 l internal volume convoluted flow path encased in a stainless steel housing (4210350B, Analytical Scientific Instruments). The connecting and exit tubing consists of 1.6 mm o.d., 0.76 mm i.d. PEEK. 3.3. Optical design Light from a 2.3 mm diameter 0.5 W tungsten lamp (4115-B, Gilway Technical Lamp) is collected by butt coupling to a 0.37 numerical aperture 1000 m fused silica fiber (ThorLabs Inc.). The fiber is fitted into the custom made PEEK z-cell (Fig. 1) using a nut (P-331, Upchurch Scientific Inc.) and ferrules (P-359, Upchurch Scientific Inc.). The cell pathlength is 1.1 cm with a 17.7 l internal volume. The fiber input and output ferrule ends create a watertight seal by compressing Teflon gaskets onto fused silica windows. The transmitted light is collected by another 1000 m fiber optic coupled to a 1 3 fiber optic bundle splitter (FiberTech Optica Inc.) (Yuan and DeGrandpre, 2006). The light from each bundle channel passes through a series of optical filters, which isolate the three wavelengths of interest (434, 578, and 780 nm). Measurements at 780 nm, where mCP has no absorbance, serve as a correction for light throughput changes between blank measurements. Additional filters are used to minimize out of band (stray) light. For detection at the 780 nm reference channel, two 780 nm bandpass filters (780-10-50; Intor, Inc.) are used. At the two analytical wavelengths, optical interference filters (435.8-10-45 and 577.7-10-50; Intor, Inc.) are combined with heat absorbing filter glass (040FG11 at 434 nm and 005FG13 at 578 nm, Andover Corp.). The combined filters reduced stray light to b 0.15% (Yuan and DeGrandpre, 2006) which is critical for good absorbance and pH accuracy. All interference filters have a 10 nm bandpass at full width half maximum; however, the center wavelength can vary slightly between filters with the same part number, requiring that eis (Eq. (3)) be measured for each SAMI. The filtered light is detected by silicon photodiodes (S2386-45K, Hamamatsu Corp). The

detector photocurrent is converted to voltage in an amplifying circuit, digitized into 12 bits and stored using a data logger (TFX11, Onset Computer Corp.). 3.4. Indicator solution The SAMI-pH and benchtop UV/Vis indicator solutions are comprised of 1.0 10 3 mol kg-soln 1 mCP (21176110G, 90% dye content, Aldrich) and 0.70 mol kg-soln 1 NaCl (S671-3, Fisher Chemicals) adjusted to a pH of 7.5 with 1.0 M HCl. The indicator concentration was tested over the range 0.020.0005 mol kg-soln 1 and it was found that 1.0 10 3 mol kg-soln 1 mCP gave the best balance between pH precision and flush times (results not shown). The influence of indicator pH is discussed below. 3.5. Measurement sequence Each measurement begins by flushing 3.75 ml of sample (75 pump pulses) through the normally open (NO) port on the valve (Fig. 1). This flushing is done to completely clear the system of indicator from the previous measurement so a blank measurement can be taken. Next, a 50 l pulse of indicator is injected into the seawater sample through the normally closed (NC) port on the valve (Fig. 1) followed by an additional 25 pump pulses (1.25 ml). Light intensity is recorded for each of the 25 sample pump pulses as the indicator peak flushes through the optical cell. The first four of these measurements are prior to arrival of the indicator and are averaged to obtain a blank value, i.e., 100% transmittance. Optical blanks are therefore recorded for every pH analysis rather than every 12 h as done in Martz et al. (2003). The standard deviation of the four blank measurements is typically b 0.0005 absorbance units. The full measurement sequence requires four min to complete. The SAMI 18 D-cell battery pack provides sufficient power for 2448 measurements or 50 d for a 0.5 h measurement frequency, 100 d for a 1 h measurement frequency, etc. 3.6. Molar absorptivity ratios (eis) To calculate the eis (Eq. (3)), molar absorptivities for the acidic and basic forms of mCP were determined at both 434 nm and 578 nm for the SAMI-pHs and the reference spectrophotometer (Varian Cary 300 Bio UV/Vis). A 0.68 mol kg 1 NaCl solution with 0.02 mol kg 1 NaOH was used for the determination of the basic L2 form of mCP. In this solution, greater than 99.995% is in the L2 form. A solution with pH of 5.5 was prepared for the HL form consisting of 0.018 mol kg 1 sodium acetate, 1.0 mol kg 1 HCl, and sufficient NaCl to set the ionic strength at 0.70 mol kg 1 (Clayton and Byrne, 1993). At pH 5.5, the presence of H2L, which has a poorly documented pKa and cannot be accurately quantified, is minimized. L2, which is present at pH 5.5, was accounted for by calculating its concentration and correcting for its molar absorptivity contribution to the final absorbance value (Martz, 2005). The correction was approximately 53% for HL578 due to the much larger L578. However, because

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HL578

M.P. Seidel et al. / Marine Chemistry 109 (2008) 1828

is small, pH is relatively insensitive to errors in this parameter (Table 1). Corrections for the other three molar absorptivities were all less than 0.3% of the measured value. It is critical to account for the ei temperature dependence (Zhang and Byrne, 1996; Gabriel et al., 2005). For example, using constant eis for a 10 C temperature change creates a pH error of 0.006 pH units. The molar absorptivity temperature dependence from 525 C was determined for each SAMI and the Cary UV/Vis. Replicate measurements were used to determine the precision and resulting uncertainty in calculated pH (Table 1). As can be seen in Table 1, uncertainty of the limits the pH accuracy to 0.0010.002 pH units. The measurement uncertainty is primarily controlled by indicator solution preparation. 3.7. Indicator pKa The pKa for mCP, determined on the total hydrogen ion concentration scale (Clayton and Byrne 1993; DOE, 1994) is: pK a 1245:69 3:8275 0:0021135 S ; T 4

where T is temperature in Kelvin and S is salinity. The pKa in Eq. (4) was determined for temperatures ranging from 293 K T 303 K and salinities from 30 S 37. It should be noted that in situ deployments may have temperatures outside of this range. We used the van't Hoff equation to extrapolate the pKa temperature dependence. The change in entropy and enthalpy were calculated using the constants A, B, and C reported by Millero (1995). 3.8. pH perturbation correction A gradient of indicator passing through the flow cell is used to correct for the indicator pH perturbation. The indicator and sample mix as they move toward the flow cell, generating a dilution curve (Fig. 2). The gradient of indicator absorbances is recorded and the pH is calculated at each point in the dilution
Table 1 The temperature dependence of the mCP molar absorptivities Species
HL434 HL578 L434 L578

SAMI-47 17104 26(t 25) 85 + (t 25) 2301 + 12(t 25) 38040 71(t 25)

SAMI-51 16911 26(t 25) 110 + (t 25) 2238 + 11.7(t 25) 37480 71(t 25)

UV/Vis 17340 26(t 25) 79 + (t 25) 2151 + 12(t 25) 37973 71(t 25)

SAMI precision 52 11 27 170

SAMI-pH error 0.0012 0.0010 0.0009 0.0020

Fig. 2. (Top) Absorbances calculated from intensity measurements taken for 25 pump pulses as the indicator slug passes through the flow cell (1 absorbance measurement obtained per pump pulse). Absorbances between 1.5 and 0.2 absorbance units are used to calculate the pH (symbols). Filled circles are A434 and open circles are A578. (Bottom) pH versus total indicator concentration [mCP]T for the SAMI data from the top figure (filled circles) and theoretical equilibrium model calculations (open circles). A linear extrapolation through each data set gives an R2 value of 0.9998 for the SAMI data and N 0.9999 for the pH perturbation model. The y-intercept (7.9169) represents the perturbation-free pH.

curve using Eq. (2). The total indicator concentration [HL]T, which is the sum of the two indicator species (Eq. (5)), is calculated from absorbances and molar absorptivities using Eqs. (6) and (7), HLT HL L2 5 HL A434 L e578 b A578 L e434 b 2 2 e HL 434 L e578 b HL e578 L e434 b 6

Molar absorptivity values are given in kg mol 1 cm 1 and temperature (t) is in C. These values are only applicable to mCP dye lot 11517KC from Sigma Aldrich. The average SAMI precision is calculated as the standard deviation of replicate measurements (n = 9) from both instruments. The SAMI-pH error is estimated by adding or subtracting the SAMI precision from the molar absorptivity value and recalculating the pH value.

2 A578 HL e434 b A434 HL e578 b : L 2 2 HL e434 L e578 b HL e578 L e434 b

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The are in kg-soln mol 1 cm 1 and pathlength (b) in cm. A plot of pH versus [HL]T is extrapolated to zero indicator concentration to determine the perturbation-free pH (Fig. 2). Mixing occurs at both the leading edge and tail end of the indicator slug as shown in Fig. 2. The leading edge of the indicator pulse has only 23 absorbance values in the optimal absorbance range (0.21.5 a.u.) (Ingle and Crouch, 1988); however, the tail end of the dilution curve is more dispersed. Approximately 9 points in this portion of the dilution curve are used to create the perturbation correction plot (Fig. 2). Because in this case the indicator pH is 7.5, the pH decreases with increasing indicator concentration. It was proposed in Martz et al. (2003) that a linear extrapolation could be used to determine the pH with no indicator present. We examined the linearity of the pH perturbation using an equilibrium model. The model includes the CO2 and indicator equilibrium expressions, equilibrium constants, the original DIC and AT of the sample and indicator, and dilution factors (calculated from the total indicator concentration). The program calculates a pH value for a range of points with varying indicator concentration and dilution factors. An example of a theoretical pH perturbation curve is shown in Fig. 2. The R2 value for a linear fit to the theoretical data is N 0.9999 over the [HL]T range used. Linear relationships are also found over the entire pH range tested (7.58.5) (not shown). Therefore, a linear fit is used with the dilution curve data from each sample measurement (Fig. 2) to determine the pH perturbation. 3.9. Laboratory tests Laboratory pH measurements were made on certified reference materials (CRMs) (Dickson, 2001) or seawater collected in Hood Canal, Washington (USA). To prevent CO2 exchange during replicate analyses, samples were stored in gas-impermeable bags (Pollution Measurement Corp.) and the bags were connected directly to the input line of the sensor (Fig. 1). All laboratory measurements were made at 20 C. Temperature was controlled to 0.05 C using a circulating water bath (Thermo Neslab RTE 7) and a water chamber placed over the top of the SAMI-pH. The sample bags were also placed inside the chamber so that the sample and optical cell of the SAMI-pH were at the same temperature. All temperature probes and water baths were calibrated to a certified digital thermometer (4400, Eutechnics). Comparison measurements were obtained using the Cary double beam UV/Vis spectrophotometer. Dual 10-cm jacketed cell holders were used for temperature control. The 10 cm sample cuvet was mixed using a magnetic stirrer (Spectrocell, Inc.). Each pH measurement took sample directly from the same bag used for the SAMI-pH measurements. The outlet tube from the bag was placed inside the cuvet and sample was allowed to overflow the cuvet 10 volumes to insure the cuvet was thoroughly rinsed. The cuvet was then placed inside the UV/Vis and allowed to reach the desired temperature ( 5 min). Each measurement used three 40 l additions of 1 10 3 mol kg 1 indicator with absorbance measurements taken for each addition.

As in the SAMI-pH, the resulting plot of pH versus total indicator concentration was used to linearly extrapolate to a zero indicator concentration. 3.10. Field tests In situ testing of SAMI-47 and SAMI-51 was performed in a seawater tank and off the SIO pier. Comparison measurements were made on a Cary 1E UV/Vis spectrophotometer. Samples were taken from the tank or pier in glass bottles with minimal head space. Samples were first placed in a water bath for 20 min set to the in situ water temperature and then a 10 cm cuvet at the same temperature was filled with sample, minimizing the head space. The cuvet was inserted into the Cary 1E for a blank measurement and indicator was added as described above but the cell containing sample and indicator was shaken by hand. All discrete pH samples were measured immediately after collection using the procedure described above. Preliminary tests were made in a 100-gallon tank filled with seawater collected from a pipeline that extends 400 m into the Pacific Ocean. A re-circulating pump was used to keep the water uniformly mixed. The SAMI-pH instruments were programmed to take measurements every 15 min. Discrete samples were collected at 1 h intervals. Immediately after these tests, the SAMI-pH instruments were deployed from the SIO pier for a three week period. Salinity, temperature, and depth were measured using a CTD (Microcat, Seabird, Inc.). Discrete samples were collected for DIC, AT, pH, and salinity. The SAMI-pHs were programmed to measure pH every 30 min. The SAMI-pH instruments went through a number of preparation steps in order to safely package them for deployment off the pier. A copper cage was placed around the exposed SAMI flow cell, fiber optics, temperature probe, and pump-valve housing in order to protect the SAMI and provide fouling resistance. The reagent bag was placed in a PVC box clamped to the SAMI housing. The SAMI was then placed in a stainless steel cage for protection and to provide solid anchor points. A 100foot serial communication cable was attached to the SAMI to monitor the instrument performance. The deployment took place from April 27, 2006 through May 25, 2006. Water depth at the end of the pier is 8 m and the instruments were placed at 4 m depth on opposite sides of the same piling (2 m apart), with a north and south orientation to obtain the same exposure to east west tidal and wave motion. 4. Results and discussion 4.1. Indicator pH The pH perturbation is strongly dependent upon the difference between the sample and indicator pH. To determine the performance sensitivity to indicator pH, a 5.0 10 3 mol kg 1 indicator solution was adjusted to pHs of 8.2, 8.0, 7.8, and 7.5 (0.1) using 1 M HCl or NaOH. Over this range, indicator pH did not significantly influence accuracy and precision using

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M.P. Seidel et al. / Marine Chemistry 109 (2008) 1828 Table 3 The pH precision for measurements made over the typical seawater range pH 7.6862 7.7833 7.9159 7.9193 8.0260 8.2029 Precision 0.0005 0.0009 0.0005 0.0006 0.0007 0.0009 95% confidence 0.0002 0.0003 0.0002 0.0002 0.0003 0.0004 Measurements 23 50 25 25 17 25

the linear extrapolation method; however, the best fits (highest R2) were found with the lowest pH solution (Table 2). Based on these results, we continued to use an indicator solution pH of 7.5, the pH chosen during the early stages of testing. Although it makes intuitive sense to use a pH close to the sample pH to minimize the perturbation, the results here suggest relative insensitivity to this parameter. 4.2. Flow cell gradient Initially, we were concerned that the gradient of indicator concentration in the SAMI-pH optical flow cell could lead to systematic errors because Beer's Law assumes a homogeneous concentration. The validity of this assumption with the 17 l optical cell was examined. The absorbance gradient between two consecutive 50 l pump pulses is estimated from data in Fig. 2. We split this absorbance gradient into 30 separate slices along the flow cell and converted the absorbances to transmittance. A corrected absorbance and pH were then calculated from the average of these thirty transmittance values. This procedure estimates a +0.0004 pH unit error in the actual pH of the sample. Therefore, the absorbance gradient is not a significant problem in the SAMI-pH; however, it could be more important for systems that use long (10 cm) pathlength cells such as liquid core waveguides (Liu et al., 2006) if the sample and indicator are not well mixed. 4.3. Laboratory precision and accuracy In the Martz et al. (2003) design, precision was found to be 0.004 pH units and was controlled by blank drift, lamp stability, and microscopic bubbles. The new design precision was quantified over a wide range of sample pH (Table 3). The average precision is 0.0007 pH units (n = 165). We attribute

Samples are CRMs and Hood Canal seawater adjusted to varying pH values using concentrated HCl or NaOH. Measurement temperature was 25 C.

the improved precision to reduced bubble trapping in the zcell, a broader mixing curve due to the static mixer, and implementation of frequent blank measurements. The accuracy for the two SAMI-pH instruments was determined by analyzing Hood Canal seawater (S =31, t =25 C) every 20 min for an 8-hour period followed by analysis of the remaining sample on the Cary UV/Vis (n =7). The average pH measured by SAMI-47 and SAMI-51 were 7.9193 0.0005 and 7.9159 0.0006, respectively. The average pH measured on the Cary spectrophotometer was 7.91800.0008 resulting in an accuracy, relative to the Cary, for SAMI-47 and SAMI-51 of +0.0013 and 0.0021, respectively (n =7). 4.4. SAMI-pH in situ testing During the initial in situ tests in the laboratory seawater tank, the average offset between SAMI-47 and SAMI-51 for 45 tankmeasurements was 0.0017 units over a pH range of 8.19 8.21 and temperature from 22.1 to 22.2 C (data not shown). Benchtop spectrophotometric pH measurements agreed to within +0.0005 for SAMI-47 and 0.0015 for SAMI-51 (n = 12). The data collected during the 22 d deployment off Scripps pier are shown in Fig. 3. A total of 1005 pH measurements were made by each SAMI. The pH was calculated using the linear extrapolation method described above and shown in Fig. 2. Sample regression plots for SAMI-47 and SAMI-51 at two different pH values are shown in Fig. 4. All in situ and sample data were corrected to the same in situ temperature using a temperature coefficient of 0.0015 pH units per 0.1 C calculated using the salinity and DIC/AT found at the site. The resulting pH ranged from 8.42 to 7.74 with changes as large as 0.6 pH units over diurnal and tidal cycles (Fig. 3). The time-series show large tidal swings and associated temperature and salinity changes (Fig. 3). The depth ranged from 3.5 to 5.5 m and temperature changes N 6 C were observed during the tidal oscillations. The temperature also increased from a daily high of 1516 C at the start of the deployment to 18 19 C at the end of the deployment. The presence of large lateral and vertical temperature gradients was confirmed using the temperature records from SAMI-47 and SAMI-51. The two SAMI-pH temperatures differed by up to 2 C during periods of rapid temperature fluctuations. These times correspond to

Table 2 Evaluation of different indicator pHs [mCP] (mol kg 1) 0.005 0.005 0.005 0.005 Indicator Average pH Slope pH pH precision (pH kg mol 1) 8.2 8.0 7.8 7.5 7.8890 7.8884 7.8886 7.8886 0.0004 0.0004 0.0004 0.0007 222 40 166 366 R2 pH perturbation

0.98 0.011 0.79 0.002 0.97 0.008 1.00 0.018

The average seawater sample pH was determined using the pH perturbation method described in the text (n = 5). Slope is the average of the five slopes generated from the pH versus total indicator plot. The R2 value is the average for the five measurements. As expected, the pH perturbation is larger (larger positive or negative slope) when the pH of the indicator is further away from the sample pH (to within the tolerance of the uncertainty in the indicator pH). The perturbation is the change in pH of the sample due to 5 10 5 mol kg 1 total indicator concentration (the indicator concentration in the indicator reservoir was 5 10 3 mol kg 1). The indicator pH was determined using a pH electrode and is only accurate to within 0.1 pH units. Measurement temperature was 25 C.

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Fig. 5. Comparison between SAMI-47 and SAMI-51 during the 22 d deployment calculated from Fig. 3 data. The average offset is 0.0042 0.0126 (n = 883). See text for more discussion.

Fig. 3. In situ SAMI-pHs, salinity, temperature and depth during the 22 d deployment off SIO pier.

significant pH differences between the SAMIs, presumably due to large pH gradients associated with the temperature (water mass) differences. Therefore, any time the measured temperature difference between SAMI-51 and SAMI-47 was N 0.2 C, the pH measurements were removed from the intercomparison between the two instruments (122 measurements were removed). Using the remaining data, the agreement between SAMI-47 and SAMI51 was 0.0042 0.0126 (n = 883) for the 22 d period (Fig. 5).

Tightening the temperature difference criteria to 0.1 C improved the comparison to 0.0040 0.011 and removed another 116 measurements. Filtering both time-series using a 2 h average further reduces the scatter between the two SAMIs to 0.0084. The lab measurements were more precise than this (Table 3) and it is likely that the large spatial gradients, particles in the flow cell (see below), and small response time differences, e.g., due to different flushing times, degraded the precision between the two instruments. The between-instrument precision is notably larger during periods when pH was rapidly changing, i.e., at the beginning and end of the deployment (compare Figs. 3 and 5). Early on, the pH rate of change was as large as 0.016 min 1 and often larger than 0.005 min 1. Looking at the pH dilution curves, SAMI-51 flushed indicator 1530 s more slowly than SAMI47 due to flow rate differences (pump efficiency), which, based on the pH rates of change, could account for a significant portion of the pH differences. Importantly, the offset is small and there is no apparent drift between the instruments during the deployment (Fig. 5). There was also no systematic trend in the SAMI difference over the pH and temperature range during the deployment (Fig. 6). These results were further verified by comparison with

Fig. 4. pH versus total indicator concentration [mCP]T for two measurements from SAMI-47 and SAMI-51 during deployment off SIO pier. The resulting pH (y-intercept) and R2 values are given in each plot.

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Fig. 6. pH difference between SAMI-47 and SAMI-51 versus temperature (A) and pH (B). The plot shows that the SAMIs respond very similarly over the range of temperature and pH found during the deployment.

measurements of pH on discrete samples collected off the pier on April 30 and May 3. SAMI-47 was deployed for both of these days while SAMI-51 was deployed on May 2 and was only compared to the pH values on May 3. Good agreement was achieved between the SAMI-pH instruments and benchtop spectrophotometric pH measurements with an average pH offset of + 0.0013 (n = 8) for SAMI-47 and 0.0043 (n = 3) for SAMI-51. After May 3, pH values were calculated from DIC and AT from discrete samples using CO2SYS (Lewis and Wallace, 1998) and the dissociation constants of Mehrbach et al. (1973) refit by Dickson and Millero (1987). Using these pH values, the average offset for both SAMI-47 and SAMI-51 are nearly identical at 0.0348 and 0.0330 pH units (n = 14). Use of other CO2 equilibrium constants in CO2SYS (Lewis and Wallace, 1998) did not improve the offset. Examination of the discrete sample data pointed to a possible error in the AT values. In surface waters, AT often follows a conservative relationship with salinity (Millero et al., 1998); however, AT of the discrete samples varied by 40 mol kg 1 while the sample salinities were very similar (33.54 0.05). Using an approximate relationship between salinity and AT for California coastal waters (AT = 2150 + 44 [S 31.25]) (G. Friederich, pers. comm., 2006), the expected AT for the salinity range observed is 2249 2252 mol kg 1 compared to the measured range of 22162254 (14) mol kg 1. Using the salinity-predicted AT and the measured DIC, the pH errors reduce to 0.014 and 0.012 pH units for SAMI-47 and SAMI-51, respectively. Uncertainty in the pK a (Eq. (4)), due to extrapolation to the in situ temperatures, may also contribute to the pH uncertainty. Although the instruments were only in the water for a short time, barnacle and algal growth completely covered the instruments. One concern in such an intense fouling environment is that, in drawing in a sample for analysis, particles can occlude the optical path or clog the plumbing. The 780 nm reference channel light throughput for the two instruments during the 22 d deployment suggest that particles may have been drawn in at times (Fig. 7). The fluctuating intensity for SAMI-51 from May 89 may be due to small particles entering the tubing and optical cell. The return to the original intensity value shows

that the particles eventually flushed out. SAMI-47 shows a different trend with the intensity slowly drifting downward with only 40% of the original intensity at the end of the deployment. The resulting behavior was most likely due to lamp drift or particulate matter entering the flow cell and slowly reducing the light throughput. No particles were evident upon inspection of the inside of the flow cell, however. The change in light throughput shown in Fig. 7 was accounted for by obtaining blank measurements for each pH analysis, i.e., for one set of data as shown in Fig. 2. To verify the need for frequent blanks, pH values for both instruments were calculated using individual blank values and an initial blank value over a 32-hour section of the field deployment data (May 2022). Using individual blank values over this time-period, the average difference between SAMI-47 and SAMI-51 pH values was 0.0017 0.0149 pH units (n = 64), while using a single initial blank value gave an average offset of 0.0070 0.0246 pH units. No drift was observed between the two instruments (Fig. 5), even with the large changes in light throughput (Fig. 7), demonstrating the robustness of the technique.

Fig. 7. % transmittance (relative light throughput) at 780 nm. This wavelength is used for the correction of lamp drift, bubbles, particles, and any other factors that affect light throughput between blank measurements. The % transmittance is calculated relative to the initial throughput at the start of the deployment.

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5. Conclusions Multiple steps were undertaken to improve the SAMIpH performance. The filter-based detection system improved the signal-to-noise and consequently, the absorbance precision. Molar absorptivities were corrected for the presence of other indicator forms by using the pH, concentration, and absorbance values of the acidic and basic solutions. With these changes, the agreement between the Cary UV/Vis and SAMI-pH instruments in a laboratory setting was improved from 0.003 to +0.0017. Optimization of various procedures, including the pH perturbation correction and mixing of the indicator and sample, improved the precision to 0.0007 compared to the previous precision of 0.004 of the Martz et al. (2003) design. The 22 d deployment off Scripps pier collected from SAMI-47 and SAMI-51 collected the longest known in situ spectrophotometric seawater pH time-series to date. The excellent agreement between the two SAMI-pH instruments and the Cary UV/Vis is very promising; however, further tests are needed to determine internal consistency with other inorganic carbon parameters. The SAMI-pH can be used for studies related to pressing contemporary carbon cycle questions such as the oceanic uptake of atmospheric CO2 and the impacts of ocean acidification. The instrument can be deployed on many different platforms including Volunteer Observing Ships, permanent moorings, drifters, autonomous profilers, and AUVs. Development of sensors for DIC and AT is needed to make possible fully autonomous characterization of the inorganic carbon system, i.e., through combined measurements of pH or pCO2 with DIC or AT. Acknowledgments We thank Cory Beatty (University of Montana) for lab and field assistance, Todd Martz (formerly UM, now at MBARI) for discussions and Charles Coughran (SIO) for deployment of instruments. Financial support was provided by NSF grant OCE-0327763 and a NSF-EPSCoR fellowship to M. Seidel. References
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