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Non-Severinghaus Potentiometric Dissolved CO2 Sensor with Improved Characteristics
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Non-Severinghaus Potentiometric Dissolved CO2 Sensor with Improved Characteristics
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Department of Inorganic and Analytical Chemistry, University of Geneva, Quai Ernest-Ansermet 30, CH-1211 Geneva, Switzerland
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Analytical Chemistry

Cite this: Anal. Chem. 2013, 85, 3, 1332–1336
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https://doi.org/10.1021/ac303534v
Published January 10, 2013

Copyright © 2013 American Chemical Society. This publication is licensed under these Terms of Use.

Abstract

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A new type of carbon dioxide sensor comprising a pH glass electrode measured against a carbonate-selective membrane electrode based on a tweezer type carbonate ionophore is presented here for the first time. No cumbersome liquid junction based reference element is utilized in this measurement. The sensor shows an expected Nernstian divalent response slope to dissolved CO2 over a wide range covering the routine environmental and physiological PCO2 levels. Unlike the conventional Severinghaus CO2 probe for which the response is substantially delayed to up to 10 min due to diffusion of carbon dioxide into the internal compartment, the ion-selective CO2 sensor proposed here shows a response time (t95%) of 5 s. When used together with a traditional reference electrode, the sensor system is confirmed to also monitor sample pH and carbonate along with carbon dioxide. A selectivity analysis suggests that Cl does not interfere even at high concentrations, allowing one to explore this type of sensor probe for use in seawater or undiluted blood samples. The CO2 probe has been used in an aquarium to monitor the CO2 levels caused by the diurnal cycles caused by the metabolism of the aquatic plants and shows stable and reproducible results.

Copyright © 2013 American Chemical Society

The importance of dissolved carbon dioxide sensing cannot be overemphasized because of the key roles it plays in the clinical, biological, environmental, and eco-biological sciences. Carbon dioxide is a poorly reactive molecule, and for this reason, receptors based on the direct molecular recognition for carbon dioxide are rarely reported. Recently, measuring methods based on infrared and nondispersive infrared spectroscopy, photoacoustic effect, surface plasmon resonance, and thermal conductivity have been increasingly being explored. (1-5) Carbonate selective membrane electrodes have been used in commercial clinical analyzers to measure the total CO2 levels of blood via adjustment of the samples to high pH (8.5 to 9.5). (6) At the same time, the conventional and widely used carbon dioxide sensors in blood gas analyzers and in environmental monitoring are still based on the Severinghaus principle. (7-9)
The Severinghaus CO2 probe was first conceived by Richard Stow in 1954 and further improved by Dr. John W. Severinghaus. (7) In 1958, it was applied to the first blood gas analysis system for the measurement of carbon dioxide partial pressure in blood. (10) Today, the Severinghaus CO2 probe is relatively inexpensive and easy to handle. Specifically, it utilizes a pH electrode in contact with a thin layer of bicarbonate solution that is separated from the sample by a gas permeable membrane such as rubber or Teflon. CO2 dissolved in the sample is able to diffuse through the membrane into the inner NaHCO3 solution where it is allowed to change the pH until CO2 partition equilibrium is established. The resulting pH value of the bicarbonate solution is then recorded and related to dissolved CO2 levels in the sample. Oceanographic CO2 measurements are typically done in the same manner but sometimes make use of a stream of dissolved pH indicator dye to report on the internal pH change in a highly reproducible fashion. (11)
Despite its wide use, some key drawbacks of the Severinghaus CO2 probe are still very difficult to overcome. In general, the use of the Severinghaus CO2 probe has been confined to the studies of media with high carbon dioxide concentration. Because it works on the basis of spontaneous diffusion and the establishment of bulk phase equilibrium between the internal compartment and the sample, the response time of the Severinghaus CO2 probe is typically 1 min or longer. This response time increases with higher inner NaHCO3 levels. (10, 12) This slow response of the Severinghaus CO2 probe has been characterized by numerical simulations and is therefore well understood. (13, 14) It is not really an adequate tool when rapid real time variations of CO2 need to be monitored, as in the depth profiling of aquatic systems or the rapid analysis of clinical blood samples.
The sensitivity of the Severinghaus CO2 probe also decreases as the carbon dioxide partial pressure drops, requiring a careful calibration in order to work at low CO2 levels. (10) Moreover, while it prevents interference from ionic solutes, the gas permeable membrane allows the penetration of other volatile acids such as H2S and HCl in their neutral form that can potentially cause interference. (12, 15-17) The analysis of total carbon balance near the sediment–water interface may be affected by the presence of H2S, resulting in an experimental bias. (18, 19)
Carrier based ion-selective electrodes (ISEs) have evolved in the past years to become a well-established routine analysis technique in clinical laboratories, process control, and environmental analysis. (20, 21) Note that many interference problems mentioned above can be avoided using a carbonate-selective electrode at the backside of a Severinghaus type CO2 probe, rather than a pH electrode, as established by Meyerhoff. (22)
We postulate here based on thermodynamic considerations (23) that a pH electrode measured directly against a carbonate electrode without the use of a traditional reference electrode gives a direct measurement of free dissolved CO2 levels. This strategy has recently been explored in some analogy for the optical detection of CO2 and has earlier been applied for the potentiometric detection of dissolved H2S. (24, 25) Any earlier attempts to develop a CO2 probe based on this attractive principle would have required a highly selective carbonate responsive membrane electrode, which was not available at the time. More recently, a new class of carbonate selective ionophore based on a molecular tweezer architecture was reported by the groups of Nam and Cha for the realization of highly selective carbonate selective ISEs. (26, 27) The optical carbon dioxide sensor mentioned above made use of this very ionophore. (24) We report here on an attractive potentiometric carbon dioxide sensing probe that does not require a gas permeable membrane and offers improved sensing characteristics for a wide range of potential applications.

Experimental Section

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Reagents

Poly(vinyl-chloride) (PVC, high molecular weight), bis(2-ethylhexyl) adipate (DOA), N,N-dioctyl-3α,12α-bis(4-trifluoroacetylbenzoyloxy)-5β-cholan-24-amide (carbonate ionophore VII), tridodecylmethylammonium chloride (TDMACl), 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris), and tetrahydrofuran (THF) were obtained from Sigma-Aldrich. Carbon dioxide (5.0 vol % in nitrogen) and nitrogen gas (99.9 vol %) were purchased from PenGas in Switzerland. Aqueous solutions were prepared by dissolving the appropriate salts in Milli-Q purified water. All solvents and reagents used were analytically pure unless otherwise specified.

Apparatus

The glass pH electrode with combined Ag/AgCl reference (Ecotrode Plus) and the double junction Ag/AgCl reference electrode were purchased from Metrohm AG in Switzerland. CO2 and N2 gas were mixed in a gas mixing chamber with a magnetic stirring bar, and the gas mixture was subsequently bubbled into the sample compartment with a specially designed sealed cap. The commercial Severinghaus CO2 probe was purchased from Lazar Research Laboratories, Inc., US. Potential responses of the electrodes were measured with an EMF-16 precision electrochemistry EMF interface from Lawson Laboratories Inc. The aquarium was prepared by planting 3 specimens of Egeria densa (E. densa) in 10 L of fresh water.

Membrane and Measurements

The carbonate-selective membranes were prepared by dissolving the mixture composed of 8.3 mg of carbonate ionophore VII, 60 mg of PVC, 2 mg of TDMACl, and 100 μL of DOA in 2 mL of THF. The cocktail solution was then poured into a glass ring (22 mm in diameter) placed on a slide glass and dried overnight at room temperature under a dust-free environment. Small disks were punched from the cast films and mounted in Ostec electrode bodies (Ostec, Sargans, Switzerland).
The calibration was done in 10–4 M NaHCO3 solution equilibrated with different partial pressures of CO2. The selectivity and response time of the sensor were characterized in 0.1 M, pH 8.0 Tris–H2SO4 buffer solutions. For the experiments shown in Figure 3, PCO2 was kept at 5% while different amounts of NaHCO3 were added to the solution and the potential response was monitored.
To evaluate the influence of sample pH to the sensor response (Figure 3), the ion-selective electrodes were placed into 0.01 M NaHCO3 solution equilibrated with 0.05 atm CO2 partial pressure. Aliquots of HCl were added to reduce the sample pH. After each addition step, the sample was allowed to re-equilibrate to the same CO2 partial pressure (PCO2 = 0.05 atm) and the sensor response was recorded.
For the CO2 measurement in the aquarium, the water solution containing 3 E. densa specimens was prepared and maintained at the same condition for the plants to stabilize before the electrodes were deployed. A 10 W incandescent light bulb was placed 10 cm above the water surface. The light was alternatively kept on for 16 h and off for 8 h while the electrodes were immersed in the aquarium and the sensor response was recorded.

Results and Discussion

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The sensor principle is presented in Scheme 1. The pH of the sample solution is monitored against a carbonate selective electrode. No potentially cumbersome liquid junction based reference element is required. The difference of potentials between the pH electrode and the carbonate-selective ISE is directly related to the dissolved CO2 and the partial pressure of CO2 in the atmosphere, which is at equilibrium with the sample solution.

Scheme 1

Scheme 1. Representation of the Ion-Selective CO2 Sensor and Its Working Principle
According to established dissociation equilibria, dissolved CO2 activity is coupled to carbonate and hydrogen ion activity through the acid dissociation equilibria as follows:(1)where Ka1 and Ka2 are the two acid dissociation constants for dissolved CO2. Note that dissolved CO2 is understood as the sum of carbonic acid and free carbon dioxide, the concentration ratio of which is constant at equilibrium. As ion-selective electrodes are established to respond to single ion activities, an electrochemical cell comprising a pH glass electrode and a carbonate-selective electrode responds to dissolved carbon dioxide quite naturally, without extra-thermodynamic assumptions:(2)where K and K′ are constants that vary with the inner solution composition of the electrodes. The constants are related through eq 1.
The CO2 response of the new gas sensing probe is shown in Figure 1. According to eq 2, a Nernstian slope for a divalent ion is expected for carbon dioxide, which was indeed observed. The calibration was performed with three different electrodes, and the mean values are plotted in Figure 1. As can be seen, the sensor is able to measure PCO2 at levels as low as 30 ppm (3 × 10–5 atm), which is sufficient for routine environmental CO2 measurements (180–300 ppm). (28, 29) The sensor response to even lower PCO2 could not be evaluated due to the difficulty in sample preparation at very low PCO2. For the Severinghaus CO2 probe, since it measures pH indirectly, a Nernstian slope is observed above 0.001 atm. Note that, at low PCO2 levels, the sensitivity becomes lower and a deviation from the ideal Nernstian response is observed. This sensitivity loss at low PCO2 region is due to deteriorating buffer capacity of the HCO3/CO2 couple as already reported by Severinghaus. (10)

Figure 1

Figure 1. Calibration curves of the sensor responses to CO2 in a 10–4 M hydrogen carbonate solution, the gas phase above 10–4 M hydrogen carbonate solution, and dry CO2 + N2 gas mixture with different levels of PCO2. The term Pθ on the x-axis is the partial pressure under standard conditions (1 atm). Shown structure is that of the tweezer-type carbonate ionophore used in the carbonate-selective membrane.

As shown in Figure 2, even at quite high PCO2 levels, it takes at least 5 min for the signal of the Severinghaus CO2 probe to stabilize. The lower the PCO2 value, the slower it gets. Previous numerical simulations have fully characterized this diffusion controlled process. (13, 14) The lagging behind in the response is not desired when real time monitoring of PCO2 is needed. Signal drifting will accumulate during the long equilibrating process and contribute also in part to the deviation from the ideal behavior of the Severinghaus CO2 probe shown in Figure 1.

Figure 2

Figure 2. Comparison of response time between the Severinghaus CO2 probe (SH) and the ion-selective CO2 sensor (PCO2) in 0.1 M, pH 8.0 Tris–H2SO4 buffer solution equilibrated with different CO2 partial pressures: (A) 0.0004 atm; (B) 0.0066 atm; (C) 0.0655 atm.

The normal mean PCO2 in arterial blood is in the range of 35–45 mmHg (0.046–0.059 atm), and a slow response time is currently tolerated for lack of a more performing sensor principle. (30) However, at lower PCO2, the diffusion of CO2 from the sample through the gas permeable membrane of the Severinghaus probe into the inner HCO3 containing solution becomes even slower and may become untolerable for practical use. Direct detection by potentiometry without the overlay of a diffusion membrane is orders of magnitude faster and may reach the tens of millisecond time scale with optimized fluidic control. (31) In batch mode, a time scale on the order of seconds is typical, which is attributed to the equilibration of the Nernst layer with the bulk solution. As shown in Figure 2 for the proposed ion-selective CO2 sensor, the potential indeed stabilizes typically within 5 s (t95%) upon change of different PCO2 values. The response time for the experiment of stepping from high PCO2 to low PCO2 conditions is shown in Figure S1 (see Supporting Information). The response in this direction is also fast, with t95% <10 s.
At high PCO2 values, the diffusion of CO2 from the sample into the inner solution might cause signal drift if the inner solution is not well buffered in carbonate, but this was not observed here. Carbon dioxide partitioning from the membrane into the sample may also result in potential drifts at very low values of PCO2. While these effects may be mainly relevant under rather specialized measurement conditions, they could be minimized by moving to an all solid state electrode design. (32)
The combination of a pH electrode, a carbonate-selective electrode, and a reference electrode allows one to monitor pH, PCO2, and carbonate concentration at the same time. Note that the measurements of pH and carbonate activity are, despite their acceptance in practice, thermodynamically less defined than the direct monitoring of PCO2 with the proposed methodology.
As shown in Figure S2, Supporting Information, when the sample solution was equilibrated after each NaHCO3 addition step with 5% CO2 (vol % in N2), the amount of dissolved CO2 is controlled by Henry’s law and thus does not change. This means that the PCO2 response from the sensor should remain the same. However, the level of dissociation of CO2 to the carbonate species depends on the alkalinity of the solution. Therefore, when the NaHCO3 concentration is increased gradually, an increase in pH is expected along with the increase of carbonate activity. All this information may now be obtained from a single experiment as shown in Figure S2, Supporting Information. Each addition of NaHCO3 is followed by the decrease in potential of the pH and carbonate-selective electrode, indicating the increase of pH and carbonate concentration. However, the observed PCO2 value, which is obtained from the difference of the two mentioned potentials, is confirmed to remain the same after the solution is brought to equilibrium with the atmosphere. Note that there is a spike of PCO2 immediately after each addition of NaHCO3. After equilibrating with the atmosphere (the equilibration time varies with the volume and effectiveness of stirring the solution), the signals come back to the same level. The ion-selective CO2 sensor is able to monitor these real time changes of the PCO2 re-equilibration process. In contrast, since the response time of the Severinghaus CO2 probe is on the order of many minutes and therefore longer than the equilibration time of the solution, it cannot access this kinetic information.
The selectivity of the ion-selective CO2 sensor is mainly limited by the selectivity of the carbonate-selective electrode, since the pH electrode is highly selective to hydrogen ions. The carbonate-selective electrode has been carefully characterized earlier by other groups. (26, 27) Here, the selectivity was revaluated under the same experimental conditions used in the previous reports (see Supporting Information). (26) As shown in Figure S3 and in agreement with previous work, interference from Cl remains highly suppressed, suggesting that the sensor may potentially work in unmodified seawater samples as well as blood samples where Cl is abundant. In addition, the interference of HS has been studied here for the first time. Hydrogen sulfide is a common component of the coastal environment generated by bacterial metabolism. (33, 34) Sulfide is found under anaerobic conditions such as the sediment covered with or inundated by seawater. When CO2 levels in these environments need to be determined, H2S will cause interference to the traditional Severinghaus CO2 probe because it is able to diffuse through the gas permeable membrane as a neutral species as well and change the pH of the inner NaHCO3 in analogy to CO2. At neutral pH, sulfides exist mainly in the form of HS and H2S. From Figure S3, Supporting Information, it is clear that HS is suppressed by 3.5 orders of magnitude. The hydrogen sulfide levels are reported to be at a maximum of 100 μmol per liter. (35, 36) On the basis of these results, one may reason that HS will not cause interference to the ion-selective CO2 sensor in such environments.
The carbonate-selective electrode is intrinsically responsive to carbonate activity in the sample. As the solution pH is lowered to below the pKa1 of CO2, free carbonate will become an ever smaller fraction of total carbonate in the sample and one expects a critical pH at which other ions such as chloride will start to effectively interfere. In Figure 3, the sample solution was equilibrated at every point with 0.05 atm CO2 while the pH of the sample was continuously lowered with HCl. The ideal PCO2 sensor response should be the same at all points as indicated by the broken line. The PCO2 detection limit is reached at pH values of around 4.8, below which value this probe is no longer recommended for determination of PCO2.

Figure 3

Figure 3. Response of the ion-selective CO2 sensor as a function of pH at a constant PCO2 of 0.05 atm. Solid lines for CO32– and PCO2 are fitted with the modified Nicolsky-Eisenman equation (20) with a logarithmic selectivity coefficient for CO32– over Cl of −6.2. The broken line indicates the ideal behavior of the sensor.

As an early example, we demonstrated that the ion-selective CO2 sensor could be used to monitor the metabolic cycles of aquatic plants. The aquarium contained specimens of E. densa planted in sand. Light was alternatively switched on for 16 h and off for 8 h. As shown in Figure S4, Supporting Information, when the light was on, the photosynthesis process of the plants consumed carbon dioxide and thus decreased the CO2 concentration in the aquarium. When light was switched off, the plants started to produce CO2 because of cellular respiration and resulted in an increase in CO2 concentration. The ups and downs in CO2 response were observed both for the Severinghaus CO2 probe and the ion-selective CO2 sensor. The ion-selective CO2 sensor showed a dissolved CO2 concentration of 85.3 ± 0.8 mmol/L toward the end of the night and 58.2 ± 0.7 mmol/L toward the end of the day. In contrast, the Severinghaus CO2 probe gave the dissolved CO2 concentration as 87.1 ± 17.1 mmol/L toward the end of the night and 60.8 ± 2.0 mmol/L toward the end of the day. While the dissolved CO2 concentration is difficult to quantify independently owing to the dynamic and spatially distributed nature of the experiment, the ion-selective CO2 sensor gave more stable and reproducible results since a stronger drift of the signals was observed for the Severinghaus CO2 probe. Nevertheless, the CO2 concentrations from the two sensors were close to each other and confirmed the ability of the sensor for such an application.
In conclusion, a potentiometric method of the use of pH and carbonate selective electrodes to measure carbon dioxide in solution and in its equilibrated atmosphere was introduced. The sensor showed the expected Nernstian response for a divalent ion with a detection limit sufficiently low for most environmental and clinical applications of interest. The response was orders of magnitude faster than for the conventional Severinghaus CO2 probe and allowed us to trace CO2 changes in real time. Further, the combined electrodes may also provide information about pH and carbonate while CO2 was measured, in which case the use of a reference electrode becomes necessary.

Supporting Information

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Additional information as noted in text. This material is available free of charge via the Internet at http://pubs.acs.org.

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Author Information

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  • Corresponding Author
    • Eric Bakker - Department of Inorganic and Analytical Chemistry, University of Geneva, Quai Ernest-Ansermet 30, CH-1211 Geneva, Switzerland Email: [email protected]
  • Author
    • Xiaojiang Xie - Department of Inorganic and Analytical Chemistry, University of Geneva, Quai Ernest-Ansermet 30, CH-1211 Geneva, Switzerland
  • Notes
    The authors declare no competing financial interest.

Acknowledgment

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The authors thank the Swiss National Science Foundation for financial support of this research.

References

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Analytical Chemistry

Cite this: Anal. Chem. 2013, 85, 3, 1332–1336
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https://doi.org/10.1021/ac303534v
Published January 10, 2013

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  • Abstract

    Scheme 1

    Scheme 1. Representation of the Ion-Selective CO2 Sensor and Its Working Principle

    Figure 1

    Figure 1. Calibration curves of the sensor responses to CO2 in a 10–4 M hydrogen carbonate solution, the gas phase above 10–4 M hydrogen carbonate solution, and dry CO2 + N2 gas mixture with different levels of PCO2. The term Pθ on the x-axis is the partial pressure under standard conditions (1 atm). Shown structure is that of the tweezer-type carbonate ionophore used in the carbonate-selective membrane.

    Figure 2

    Figure 2. Comparison of response time between the Severinghaus CO2 probe (SH) and the ion-selective CO2 sensor (PCO2) in 0.1 M, pH 8.0 Tris–H2SO4 buffer solution equilibrated with different CO2 partial pressures: (A) 0.0004 atm; (B) 0.0066 atm; (C) 0.0655 atm.

    Figure 3

    Figure 3. Response of the ion-selective CO2 sensor as a function of pH at a constant PCO2 of 0.05 atm. Solid lines for CO32– and PCO2 are fitted with the modified Nicolsky-Eisenman equation (20) with a logarithmic selectivity coefficient for CO32– over Cl of −6.2. The broken line indicates the ideal behavior of the sensor.

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