Anal. Chem.

2000, 72, 5338-5347

Measurement of Atmospheric Hydrogen Peroxide and Hydroxymethyl Hydroperoxide with a Diffusion Scrubber and Light Emitting Diode-Liquid Core Waveguide-Based Fluorometry
Jianzhong Li and Purnendu K. Dasgupta*

Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas 79409-1061

We describe a new automated instrument for measuring gas- and aqueous-phase H2O2. The chemistry relies on the hematin-catalyzed oxidation of nonfluorescent thiamine to fluorescent thiochrome by H2O2; this reaction is 25-fold more selective for hydrogen peroxide than its nearest alkyl hydroperoxide congener, CH3HO2. The optical characteristics of the fluorescent product are such that it is ideally excited by newly available GaN-based light emitting diodes emitting in the near-UV. A stable longlife miniature flow-through fluorescence detector based on a transversely illuminated liquid core waveguide is thus used for this purpose. The limit of detection (LOD, S/N ) 3) for liquid-phase H2O2 is 11 nM. A temperaturecontrolled high-efficiency Nafion membrane diffusion scrubber is used to collect gaseous hydrogen peroxide with near-quantitative efficiency with an S/N ) 3 LOD of 13.5 pptv. The system responds to hydrogen peroxide and hydroxymethyl hydroperoxide but not to methyl hydroperoxide. The use of very inexpensive and stable reagents, highly sensitive detection, benign chemistry, and a fluorescence detector using a solid-state illumination source results in a particularly affordable automated instrument. Design and performance details and illustrative results from a 1999 field campaign (Atlanta Supersite Study) are presented.
Hydrogen peroxide is an important analyte in clinical chemistry. It also plays a central role in atmospheric studies due to its ability to oxidize dissolved SO2.1 Hydrogen peroxide, methyl hydroperoxide (MHP), and hydroxymethyl hydroperoxide (HMHP) are of particular interest and detailed reviews are available.2 In the aqueous phase, H2O2 is the primary analyte of interest since MHP displays a much lower Henrys law constant3 and HMHP T H2O2 + HCHO exhibits a dissociation constant of 6 mM at 25 C.4 Luminol chemiluminescence (CL) can be very sensitive and has been exploited for the measurement of marine
* Corresponding author: (e-mail) sandy.dasgupta@ttu.edu. (1) Navas, M. J.; Jimenez, A. M.; Galan, G. Atmos. Environ. 1999, 33, 22792283. (2) Gunz, W. G.; Hoffman, M. R. Atmos. Environ, 1990, 24A, 1601-1634. Jackson, A. V. Crit. Rev. Environ. Sci. Technol. 1999, 29, 175-228. (3) OSullivan, D. W.; Lee, M.; Noone, B. C.; Heikes, B. G. J. Phys. Chem. 1996, 100, 3241-3247. (4) Zhou, X.; Lee, Y.-N. J. Phys. Chem. 1992, 96, 265-272.

H2O2.5 Many oxidants generate luminol CL; direct measurement of H2O2(g) by luminol CL is difficult. Alternative CL techniques for H2O2(g) have been developed.6 Few have attempted to measure gaseous peroxides electrochemically7 because it is not sufficiently sensitive. Introduced by Lazrus et al.,8 fluorometry dominates present atmospheric peroxide measurements. There are pedagogical merits to fluorometric H2O2 measurements as well.9 Typically, a nonfluorescent substrate is oxidized by H2O2 to a fluorescent product mediated by a peroxidase enzyme, typically horseradish peroxidase (HRP).8-10 Cost and stability issues of HRP have led to attempts to use photocatalysis11 and other nonenzymatic reactions.12 Synthetic metalloporphyrins have been extensively studied as HRP mimics13 and shown to be effective.14 Phthalocyanine complexes are effective catalysts in both acidic and basic solutions.15 Several Fe-porphine compounds, inexpensive commercial products from animal blood, display peroxidatic activity.14 Bovine hematin displays a greater peroxidatic activity per unit mass than any commercial HRP preparation at <0.2% of the cost.14 Hematin-based H2O2(g) measurement systems were reported.16 For the hematin-H2O2 system, 14 potential fluorogenic substrates were screened (including the best candidates earlier identified by Guilbault et al.17) and 4-hydroxyphenylpropanol was the best performer while p-cresol provided 80% of the best
(5) Price D.; Mantoura, R. F. C.; Worsfold, P. J. Anal. Chim. Acta 1998, 371, 205-215. Yuan, J.; Shiller, A. M. Anal. Chem. 1999, 71, 1975-1980. (6) Stigbrand, M.; Karlsson, A.; Irgum, K. Anal. Chem. 1996, 68, 3945-3950. (7) Huang, H.; Dasgupta, P. K. Talanta 1997, 44, 605-615. Holmstrom, S. D.; Cox, J. A. Electroanalysis 1998, 10, 597-601. (8) Lazrus, A. L.; Kok, G. L.; Gitlin, S. N.; Lind, J. A. Anal. Chem. 1985, 57, 917-922. (9) Weinstein-Lloyd, J. B.; Lee, J. H. J. Chem. Educ. 1995, 72, 1053-1055. (10) Jacob, P.; Klockow, D. Fresenius Z. Anal. Chem. 1993, 346, 429-434. (11) Genfa, Z.; Dasgupta, P. K.; Edgemond, W. S.; Marx, J. N. Anal. Chim. Acta 1991, 243, 207-216. (12) Lee, J. H.; Tang, I. N.; Weinstein-Lloyd, J. B. Anal. Chem. 1990, 62, 23812384. (13) Saito, Y.; Nakashima, S.; Mifune, M.; Odo, J.; Tanaka, Y.; Chikuma, M.; Tanaka, H. Anal. Chim. Acta 1985, 172, 285-287. Saito, Y.; Mifune, M.; Nakashima, S.; Odo, J.; Tanaka, Y.; Chikuma, M.; Tanaka, H. Talanta 1987, 34, 667-669. (14) Zhang, G.; Dasgupta, P. K. Anal. Chem. 1992, 64, 517-522. Howell, R. R.; Wyngaarden, J. B. J. Biol. Chem. 1960, 235, 3544-3550. (15) Chen, Q.-Y.; Li, D.-H.; Zhu, Q.-Z.; Zheng, H.; Xu, J.-G. Anal. Lett. 1999, 32, 457-469. (16) Zhang, G.; Dasgupta, P. K. Anal. Chim. Acta 1992, 260, 57-64. (17) Guilbault, G. G.; Brignac, P. J., Jr.; Juneau, M. Anal. Chem. 1968, 40, 12561263. 10.1021/ac000611+ CCC: $19.00 2000 American Chemical Society Published on Web 09/27/2000

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fluorescent intensity, at 1% of the cost. However, all of above schemes require excitation between 300 and 326 nm. Narrowband GaN-based light emitting diodes (LEDs) emitting at375 nm are now available;18 and simple liquid core waveguide (LCW)based fluorescence detectors have been described.19 A LCW-based solid-state fluorometer for H2O2 will be attractive. With metallophthalocyanine or hemin as a catalyst, thiamine (vitamin B1), can be used to determine H2O2.15,20 It is oxidized to fluorescent thiochrome with an excitation maximum in our desired range.21,22 Generally, the analytical methods above do not discriminate between H2O2 and the two principal organic peroxides, MHP and HMHP. The enzyme catalase or a MnO2 catalyst has often been used to selectively destroy H2O2 and thus establish differential measurements.8,23 Both approaches have problems. Extending the data obtained with pure peroxides produces errors with mixtures of H2O2 and organic peroxides in the catalase system.24 With MnO2, there may be some destruction of organic peroxides.25 Our experience with continuously operating MnO2 reactors indicates that they lose activity and must be replaced on a frequent basis, requiring recalibration. In a different approach, the Fenton reaction has been used to measure H2O2 + HMHP (40%) + MHP (12%) with a low-pH collector, H2O2 + HMHP + MHP (12%) with a highpH collector, and H2O2 + HMHP + MHP (60%) with a high-pH collector.26 All gaseous H2O2 measurement techniques that involve aqueous scrubbing have some degree of O3 interference since O3-water reactions at surfaces produce H2O2.27 Such reactions are facilitated at a higher pH,28 and the extent of O3 interference with a high-pH collector needs careful investigation. At the present time, the best method for the determination of individual peroxides appears to be chromatography with postcolumn reaction-fluorometry.29 Measurement of peroxide gases by such techniques must nevertheless involve collection. With aqueous scrubbing, collection of MHP is incomplete, albeit subquantitative collection can be calibrated for.30 On the other hand, cryogenic trapping is quantitative but can lead to significant amounts of artifact peroxides.31 Interference from SO2 poses a problem in both methods and an universally applicable solution is not available. Clearly, intercomparisons are necessary to clarify the strengths and weaknesses
(18) www.nichia.com. (19) Dasgupta, P. K.; Zhang, G.; Li, J.; Boring, C. B.; Jambunathan, S.; Al-Horr, R. Anal. Chem. 1999, 71, 1400-1407. (20) Zhu, Q.-Z.; Li, Q.-G.; Lu, J.-Z.; Xu, J.-G. Anal. Lett, 1996, 29, 1729-1740. (21) Guo, X.-Q.; Xuo, J.-G.; Wu, Y.-Z.; Zhao, Y.-B.; Huang, X.-Z.; Chen, G.-Z. Anal. Chim. Acta 1993, 276, 151-160. (22) Jie, N.; Yang, D.; Zhang, Q.; Yang, J.; Song, Z. Anal. Chim. Acta 1998, 359, 87-92. (23) Dasgupta, P. K.; Hwang, H. Anal. Chem. 1985, 57, 1009-1012. (24) Heikes, B. G. University of Rhode Island, personal communication, 2000. (25) de Serves, C.; Ross, H. B. Environ. Sci. Technol. 1993, 27, 2712-2718. (26) Lee, J. H.; Tang, I. N.; Weinstein-Lloyd, J. B.; Halper, E. B. Environ. Sci. Technol. 1994, 28, 1180-1185. (27) Heikes, B. G. Atmos. Environ. 1984, 18, 1433-1445. (28) Staehelin, J.; Hoigne , J. Environ. Sci. Technol. 1982, 16, 676-681. (29) Hellpointner, E.; Ga b, S. Nature 1989, 337, 631. Kurth, H.-H.; Ga b, S.; Turner, W. V.; Kettrup, A. Anal. Chem. 1991, 63, 2586-2588. Hewitt, C. N.; Kok, G. L. J. Atmos. Chem. 1991, 12, 181-194. Kok, G. L.; McLaren, S. E.; Staffelbach, T. A.; J. Atmos. Ocean. Technol. 1995, 12, 282-289. Sauer, F.; Limbach, S.; Moortgat, G. K. Atmos. Environ. 1997, 31, 1173-1184. Wang, K.; Glaze, W. H. J. Chromatogr. 1998, 822, 207-214. (30) Lee, M.; Noone, B. C.; OSullivan, D.; Heikes, B. G. J. Atmos. Ocean. Technol. 1995, 12, 1060-1070. (31) Staffelbach, T.; Neftel, A.; Dasgupta, P. K. Geophys. Res. Lett. 1995, 22, 2605-2608.

of the different methods. Even if only H2O2, HMHP, and MHP are present, it is minimally desirable to be certain how much of what precisely a given procedure actually measures; the response coefficients must not depend on the exact composition of the sample. Peroxide gases have also been collected with the coil,8 the diffusion scrubber32 (DS), the mist chamber, and, most recently, the condensate trap.33 The condensate trap requires no power; ice-filled test tubes are hung in a container where peroxides in the ambient air condense on the outer walls. It requires careful monitoring of humidity and is inapplicable at low absolute humidities. The mist chamber is an efficient collector but obligatorily collects the aerosol. Interaction with the deposited aerosol can lead to the loss of H2O2; interaction between ozone and deposited organic aerosol can lead to the generation of H2O2. Aerosol deposited in the sampling line has the same effect. Periodic cleaning of all contact surfaces is necessary for this reason. The DS permits simple strategies to preconcentrate.34 The extent of aerosol deposition in a properly designed DS is very low,35 but periodic routine cleaning is still necessary. The coil, probably the most widely used, shows a modest amount of aerosol loss. Insoluble particles can be carried by the liquid stream and be problematic,25 but they do not accumulate in the collection surface. Direct intercomparisons of collection techniques have not been frequent. In one such study on a coil-DS comparison, the authors concluded that the DS is superior.25 The DS used in the present work is of the straight inlet design and is a nearquantitative collector of H2O2 over a significant range of flow rates. In the present report, we describe a fully automated instrument that thus combines several chemical, material science, and technological developments since our last effort on gaseous H2O2,16 to collect H2O2 nearly quantitatively and measure [H2O2] + [HMHP]. Illustrative results from an extensive field study in the summer of 1999 in Atlanta (the Atlanta Supersite Study36) are also presented. EXPERIMENTAL SECTION Reagents. Hematin (from bovine blood, Aldrich) stock solution was prepared by dissolving 10 mg of hematin in 100 mL of 0.1 M NaOH. Refrigerated at 4 C, this solution is stable for at least 2 months. Thiamine stock solution (10 mM) was prepared by dissolving 337 mg of thiamine hydrochloride (Sigma) in 100 mL of water. Refrigerated, this solution was stable for at least 1 months. Working hematin (nominally 10 M) and thiamine solutions (100 M) were prepared daily by diluting 6.3 mL of the hematin stock to 100 mL with phosphate buffer (50 mM K2HPO4 adjusted to pH 12.0 with 2 M NaOH) and diluting the thiamine stock with water, respectively. Hydrogen peroxide stock solution (1 M) was prepared by dilution of 30% solution (Fisher) with water and standardized by titration with secondary standard KMnO4. MHP was a gift from researchers at the National Center for Atmospheric Research. HMHP was synthesized by the reaction
(32) Dasgupta, P. K.; Dong, S.; Hwang, H.; Yang, H.-C.; Genfa, Z. Atmos. Environ. 1988, 22, 949-963. (33) Deforest, C. L.; Kieber, R. J.; Willey, J. D. Environ Sci. Technol. 1997, 31, 3068-3073. (34) Genfa, Z.; Dasgupta, P. K.; Dong, S. Environ. Sci. Technol. 1989, 23, 14671474. Trapp, D.; De Serves, C. Atmos. Environ. 1995, 29, 3239-3243. (35) Zhang, G.; Dasgupta, P. K.; Cheng, Y.-S. Atmos. Environ. 1991, 25A, 27172729. (36) http://www-wlc.eas.gatech.edu/supersite/.

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Figure 1. (a) Instrument schematic. Key: P, peristaltic pump; DS, diffusion scrubber; L1, thiamine mixing coil; L2, main reaction coil, thermostated at 30 C; F, flush port (normally closed); AP, air aspiration pump; FM, flowmeter; N, 23-gauge hypodermic needle (supplementary flow); V, three-way solenoid valve controller by timer T; C, activated carbon column. (b) Transversely illuminated fluorescence detector schematic. Key: T, entrance PEEK tee; LI, liquid inlet; FO, silica fiber optic; OF, colored plastic optical filter; PMT, miniature photomultiplier tube; AF, Teflon AF 2400 liquid core waveguide; SSJ, stainless steel jacket, LED. UV emitting light emitting diode; PD, photodiode for monitoring source light intensity; U, outlet union; W, waste.

of H2O2 and HCHO.37 The material was primarily an oil of HMHP with some H2O2 and bis-HMHP (BHMHP). The H2O2 present in the MHP or HMHP/BHMHP solution was removed by passing the solution through a MnO2 column,23 and the concentration of the organic peroxide in the effluent was determined by measuring the absorbance of I3- at 352 nm produced by the oxidation KI in the presence of acetic acid under a blanket of CO2. Analytical System. The arrangement of liquid and gas flows in the system is schematically shown in Figure 1a. Water was aspirated by a peristaltic pump (P, Rainin Dynamax) through the diffusion scrubber (DS), which collects the H2O2 from the gas phase to the water stream. The aqueous effluent from the DS was mixed with the thiamine reagent in a Serpentine-II knit38 PTFE reactor (L1, 0.65 mm i.d. 400 mm). The hematin stream was then merged in and the mixed stream and reacted in another similarly knit PTFE reactor (L2, 0.65 mm i. d. 1700 mm, residence time 180 s) and then flowed through the LCW-based fluorescence detector. Reaction coil L2 is potted in a low-melting bismuth-tin alloy (Cerrobase 5550-1, Canfield Technologies, Sayreville, NJ) in a poly(vinyl chloride) container containing two flexible siliconized heaters (Watlow, St. Louis, MO) in series (22.5 W total). A 100- platinum RTD monitors the temperature, and temperature control is achieved by a miniature temperature controller (type CN 1632 GNR, Omega Engg., Stamford, CT). Unless otherwise stated, the reactor was maintained at 30 C. A flush port F is provided for the detector via a tee-arm to remove recalcitrant bubbles or to wash and clean the detector (vide infra). A miniature air pump (AP, UNMP30KNI, KNF Neuberger, Trenton, NJ) was used to aspirate the sample gas through the DS. The pump outlet was connected to a three-way valve V (MBD
(37) Marklund, S. Acta Chem. Scand. 1971, 25, 3517-3531. (38) Curtis, M. A.; Shahwan, G. J. LC-GC Mag. 1988, 6, 158-164.

002, Honeywell) via flowmeter FM. This valve was controlled by an independent programmable timer (RS4A22, SSAC Inc., Baldwinsville, NY). In one position, the pump exhaust was vented, and in the other position, the pump exhaust was passed through a column I filled with pelletized activated carbon (8 mesh, Fluka) to remove any H2O2 in it and feed this zero gas to the DS. The total aspiration by AP constituted that through the DS, plus a small amount (2% of the total; total flow was typically 1.6 standard liters per minute, SLPM) through a 23-gauge hypodermic needle (N) that was connected to the AP inlet by a tee. This design ensures that when valve V is switched to provide zero gas to the DS inlet, the airflow is sufficient and no ambient air is drawn through the sample inlet; rather, a small amount of zero air back-flushes the sample inlet line. Unless otherwise stated, valve V was programmed to sample the test gas for 2 min and zero gas for 8 min. LCW Fluorescence Detector. The structure of the transversely illuminated LCW fluorescence detector is shown in Figure 1b; it is similar to previous reports19,39 except that an LED emitting in the near-UV was used as the light source. An LED excitation source is very stable, exhibits long life, and produces no heat. The lateral distance between the excitation source and the detector receiving fiber is 45 mm. Although the illuminated volume is not of major concern to this work, it is sufficiently small for the device to function as a detector in high-performance liquid chromatography. Large-core (1.5 mm) acrylic optical fiber collects the emitted light from the end of the LCW AF (1.25 mm i.d.; Random Technologies, San Francisco, CA) and conducts it to miniature photomultiplier tube (PMT; Hamamatsu H5784). As an excitation filter, we used two disks of blue plastic OF (type 861, Edmund Scientific. Gloucester, NJ). The excitation light status is monitored by a silicon photodiode (PD; BPW 34, Siemens AG).
(39) Li, J.; Dasgupta, P. K.; Zhang, G. Talanta 1999, 50, 517-623.

5340

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Figure 2. Diffusion scrubber and housing. Key: U1, U2, U3, PVDF 1/4-1/8-in. tubing union fittings; SI, sample inlet; ZA, zero air inlet; AP, to air pump; T1, T2, T3, PVDF tees; E, PVDF elbow; N, Nafion tube; J, PTFE jacket; SS, stainless steel tubes; WI/WO, water inlet/outlet (PTFE capillaries); RTD, platinum resistance thermometer connected to miniature three-wire jack M; PF, Plexiglas frame; C, filler cap; H, heater (top and bottom); HL, heater leads.

Diffusion Scrubber. A new robust design of the diffusion scrubber was developed for this work and is shown in Figure 2. The DS was housed in a Plexiglas enclosure with inside dimensions being 21/8 in. wide 201/8 in. long 1 in. deep. Flexible siliconized heater elements (2 20 in., Watlow, 50 W at 110 V) were glued by silicone rubber adhesive to the interior surfaces of both the top and the bottom covers. The main DS unit constitutes of Nafion tube N (wet dimensions 0.75 mm i.d., 0.90 mm o.d.) housed in a PTFE jacket tube J (3.2 mm o.d., 2.7 mm i.d.) held in place by poly(vinyledene fluoride) (PVDF) tees T1 and T2. T1 is connected to 1/8-1/4-in. union fitting U1 with a minimum of PTFE tubing to provide 1/4-in. tube connections to the air pump. T2 is connected with a minimum of tubing to tee T3 that connects to the sample inlet fitting U2 and to an elbow E by rigid stainless steel tubing SS. E is further connected to zero air inlet U3 by another piece of stainless steel tube SS. Nafion tube N contains inserted PTFE capillary tubing at its termini; these are sealed at the tee ports of T1 and T2 with suitable sleeves. The PTFE capillaries exit through the frame through suitable sleeves and 10-32 threaded compression seals. The active length of the Nafion membrane is 45 cm. A 100- platinum RTD was placed on the midsection of the DS jacket and affixed in place with PTFE tape. The RTD was connected in a three-wire configuration to a stereo mini-Jack M. This was connected by an appropriate cable to a temperature controller (of the same type as used with reactor L2). The heater leads were connected in series to reduce the total heater power to 25 W. We investigated a number of options for the filler material in the DS housing. The desirable characteristics were low weight, good thermal conductivity, low off-gassing of any interfering vapor, and low cost. We chose a decorative gravel for filling aquaria, available in most pet stores. The filling material can be put in or removed through a hole normally capped by a cap C. Scrubber collection efficiency was determined by connecting two identical diffusion scrubbers in series and comparing the ratio of the observed signal in the upstream versus the downstream

scrubber. The scrubber positions are then reversed, and the same evaluation is repeated. This procedure does not need to assume identical collection efficiencies of the two DS units, and the basic theory has been previously described.32 Instrument Calibration and Standard Gas Generation. Both gas-phase and liquid-phase calibrations are of interest because the instrument can be used for either gas-phase measurement or for the measurement of dissolved H2O2 in rain or cloudwater. Both types of calibration were therefore conducted. The loop-type six-port injector valve (I, Figure 1a, P/N V540, Upchurch Scientific, Oak Harbor, WA) was provided with a loop of 50-L volume for the purposes of liquid-phase calibration. The gas-phase H2O2 generation source was set up and calibrated as previously reported; the results were in good agreement with previously published Henrys law constant for H2O2.40 Gaseous MHP was generated in a similar manner using an MHP solution as the generation source solution. To inhibit the hydrolysis of MHP, the solution was prepared in 1 mM HCl. The gas-phase MHP concentration was calculated from the known Henrys law constant3 and the measured aqueous MHP concentration. Several factors complicate similar experiments with HMHP. First, the stability of dilute HMHP solutions over a long period is a concern.4 Second, the solution is not pure and selective transfer of the less soluble H2O2 to the gas phase will alter the nature and concentration of the gas-phase peroxide(s) over a period of time. We chose to perform the HMHP experiments by depositing 20 L of a HMHP solution in a Teflon tube through which 3.5 L/min air was flowing. The gas was sampled by the instrument with zero on/off cycling disabled, the area under the entire response peak was integrated to assess the response, and the results were compared with an identical experiment with hydrogen peroxide solutions. Ozone was generated by a high-pressure mercury lamp. The concentration of the generated O3 was determined by neutral
(40) Hwang, H.; Dasgupta, P. K. Environ. Sci. Technol. 1995, 19, 255-258.

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buffered potassium iodide method (1% KI buffered with phosphate, pH 6.8 ( 0.2) and measuring the absorbance of produced I3- at 352 nm. A diode array spectrophotometer (Hewlett-Packard 8453) was used for all spectrophotometric measurements. A photometric ozone monitor (model 1003 AH, Dasibi Instrument Corp.) was used as a secondary instrument to monitor the ozone concentration. The generated ozone was scrubbed with water to remove H2O2 potentially present in the ozone stream. The scrubbed ozone was diluted with a H2O2-bearing stream of known concentration for testing. Sulfur dioxide was generated with a Henrys law-based source with a source solution containing 5 mM NaHSO3 in a 50 mM potassium acid phthalate buffer at pH 4 and 20 C. The anticipated primary source concentration was 22.3 ppm, based on published equilibrium constants. The measured value (sampled into bubbler containing dilute H2O2 and ion chromatographic measurement as sulfate) showed an initial value of 18.7 ppm; that decreased to 18.0 ppm by the end of the interference studies. Field Studies. The instrument was deployed in the Atlanta Supersite experiment36 from August 3, 1999 to August 29, 1999. The site, located in midtown Atlanta (latitude 33.777 N, longitude 84.414 W, altitude 265 m MSL) provided an air-conditioned shelter for the instrument. A Teflon-coated aluminum cyclone (designed for a 2.5-m cut point at a flow rate of 3 L/min, URG, Chapel Hill, NC) with a downward pointing inlet 2 m above the roof of the shelter (6 m above ground) was used. The inlet conduit from the cyclone to the DS was PFA Teflon (5 mm in i.d.). The total flow through the sampling system was 2.7 SLPM, 1.6 SLPM sampled by the present instrument and 1.1 SLPM sampled by a second instrument designed to measure formaldehyde. The residence time in the sampling line was 2 s. The sampling line was tightly wrapped with aluminum tape to prevent exposure to sunlight. The aluminum tape was also grounded; in our experience this reduces aerosol deposition. The instrument sensitivity was checked daily with aqueous standards. The cyclone and the entire sample inlet line was washed weekly with deionized water and dried with compressed nitrogen to remove any deposited particles. The instrument was then calibrated fresh with a standard gaseous H2O2 source every week. Any calibration shift was compensated for by linear interpolation of the calibration within the interval. RESULTS AND DISCUSSION Spectral Characteristics. Figure 3 shows the spectral details relevant to this system. Thiochrome, the oxidation product of thiamine, absorbs in a broad band (fwhm 52 nm) centered at 373 nm, which is ideally excited by the narrow-band (fwhm 12 nm) GaN LEDs (NSHU 590E, Nichia America Corp.) with its emission centered at 375 nm. The emission of the product occurs maximally in a broad band (fwhm 74 nm) centered at 440 nm. The reaction medium containing the hematin catalyst itself acts as a filter toward stray excitation light. Doubtless this stray light filtering will be even more efficient if the distance between the light source and the receiving fiber is increased. Presently two sheets of a colored plastic, with a transmittance window centered near the emission maximum of thiochrome (see Figure 3) suffice as emission filters. Optimization of Reaction Conditions. Choice of Buffer and pH. These studies were carried out with aqueous H2O2 as the
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Figure 3. Spectroscopic details relevant to the measurement system. (a) Excitation spectrum of thiochrome (em 440 nm); (b) emission spectrum of GaN LED; (c) emission spectrum of thiochrome ((ex 370 nm); (d) absorbance due to the reaction medium, 5-cm path length, right ordinate; (e) absorbance of 1 disk of No. 861 colored plastic as emission filter.

test analyte. Buffering systems based on carbonate, ammonia, and phosphate were tested. An anmmoniacal medium provided the highest net sensitivity but also produced a higher blank background. While the difference is not dramatic, the sensitivity was the poorest with carbonate. We also found that, in continued use, hematin or some insoluble product therefrom slowly deposits in the system, including on the optical components. When ammonia or carbonate buffer was used, a significant decrease in the signal over 1 day was seen. With a phosphate buffer, the decrease in the signal over 24 h of continuous use was <5%. Phosphate buffer was therefore chosen. The fluorescence detector was washed with 4 mL of 2 M HCl every day to avoid any accumulation of hematin on the optical components. The pH dependence of the observed signal (Figure 4a) is a function of both the dependence of the reaction rate on pH and the dependence of the fluorescence intensity of the thiochrome formed on pH. We allowed thiochrome to be fully formed in a test tube reaction and tested the pH dependence of the fluorescence intensity of the product formed. Thiochrome was found to become fully fluorescent at a pH of g8. The pH dependence observed in Figure 4a therefore originates entirely in the dependence of the rate of production of thiochrome as a function of pH. Although different plateau values are observed, all three buffers show the same sigmoidal dependence on pH; the active form of the catalyst apparently has a pKa of 11. A phosphate buffer of pH 12 was henceforth used. The effect of the concentration of the phosphate buffer on the observed signal was also studied. The fluorescence intensity increased with increasing concentration at a constant pH of 12 but the increase was very marginal over a phosphate concentration greater than 50 mM. A 50 mM buffer concentration was henceforth used.

Figure 4. (a) Effect of the nature of the buffer and pH on net fluorescence signal; (b) effect of hematin concentration on the net signal and background.

Figure 5. (a) Effect of thiamine concentration on a 10 M H2O2 signal and background fluorescence; (b) net signal from a 10 M H2O2 injection and background as a function of flow rate.

Effect of Hematin Concentration. The results of this study are shown in Figure 4b. The fluorescence signal from H2O2 sample injections increased with increasing hematin concentrations, becoming constant at concentrations of g10 M; this was used in further work. Because of the optical filtering of the excitation light by hematin (Figure 3), the blank background decreases continuously with increasing hematin concentration. Effect of Thiamine Concentration. For 10 M H2O2 as sample, the fluorescence signal increased with increasing thiamine concentration up to 30 M, decreasing slightly thereafter and becoming constant (Figure 5a). The blank also increased slowly with increasing thiamine concentration. To operate in the constantsensitivity region, we chose a thiamine concentration of 100 M. Effect of Reaction Temperature. In the tested range from 24 to 40 C, the fluorescence signal due to H2O2 injection decreased with an increase in temperature, decreasing by 10% from 30 to 40 C. Whether this is caused by a decrease in the quantum efficiency of thiochrome or in the catalytic activity that leads to its

production, or enhancement of the production of an alternative product, nonfluorescent thiamine disulfide,41 is not known. For practical purposes, the temperature of the reactor was controlled at 30 ( 0.5 C. Effect of Flow Rate. The liquid flow rates control the residence/ reaction time and, as such, have a significant effect on the observed signal (Figure 5b). Even though low flow rates increase the net signal, the blank background is also increased and thus increases the noise level. Low flow rates also increase the response time. As a compromise between system response time and sensitivity, a flow rate of 80-100 L/min (per channel) appears to be optimum, corresponding to a total residence time of 2.53.1 min. Effect of Gas Sampling Rate on H2O2(g) Collection Efficiency. The collection efficiency of diffusion-based collectors decreases with increasing flow rate. The collection efficiencies of two nominally
(41) Puiz, T. P.; Lozano, C. M.; Tomas, V.; Ibarra, I. Talanta 1992, 39, 907912.

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Table 1. Results from SO2 Interference Study concentration, ppbv H2O2 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 2.0 2.0 3.0 3.0 4.0 4.0 5.0 5.0 SO2 0 25 50 75 150 200 0 150 0 150 0 150 0 150 0 150 signal, mV 441.3 ( 2.0 426.1 ( 6.1 400.0 ( 2.0 381.7 ( 3.9 347.4 ( 1.6 334.5 ( 1.5 434.7 ( 8.8 366.4 ( 2.4 818.3 ( 11.3 751.7 ( 5.0 1219.4 ( 1.7 1125.5 ( 10.4 1685.0 ( 13.8 1576.5 ( 8.6 2200.7 ( 36.3 2093.5 ( 25.9 % interference control -0.034 -0.094 -0.135 -0.213 -0.242 control -0.157 control -0.081 control -0.077 control -0.064 control -0.049 interference equiv pptv H2O2/ppbv SO2 -1.38 -1.87 -1.35 -1.42 -1.21 -1.05 -1.09 -1.54 -1.72 -1.62

identical DS units (A, B) were determined over a sampling rate range of 0.76-2.94 SLPM. (Note that our local atmospheric pressure is 680 mmHg and the laboratory temperature is 23 C; if the same experiments were conducted at sea level at 20 C, the same linear sampling velocities (on which collection efficiencies ultimately depend)42 will correspond to flow rates of 0.86-3.32 SLPM.) The observed collection efficiencies fi (from 98 to 88% at the highest to lowest flow rate studied) obeyed theoretical expectations in that ln(1 - fi) was linearly related with the reciprocal of the sampling rate:43

ln(1 - fA) ) -(1.8276 ( 0.0543)/Q - 1.4909 ( 0.04298, r2 ) 0.9965 (1) ln(1 - fB) ) -(1.9294 ( 0.0613)/Q - 1.4990 ( 0.04853, r2 ) 0.9960 (2)
where fA and fB are fractional collection efficiencies pertaining to DS units A and B and Q is the sampling rate in SLPM. Because the collection efficiency is high, the overall signal changes nearly linearly with the sampling rate. As an optimum choice between keeping the sensitivity high and achieving nearly quantitative collection efficiency, we selected a sampling rate of 1.6 SLPM; this permits a collection efficiency of 94% in our laboratories and 95% at the higher atmospheric pressures in the field experiments in Atlanta. Interference Studies. Sulfur Dioxide. Sulfur dioxide is highly water-soluble and is readily oxidized by H2O2 and, as such, presents a significant potential negative interference for the measurement of H2O2. Thus far, the primary effort to minimize SO2 interference has been pH control of the collection liquid and the addition of HCHO to the collection liquid to tie up the S(IV) before it can be oxidized by H2O2.26,30 The HCHO concentration in the collection liquid and the pH are both important considerations. Addition of HCHO at pH levels of g6 leads to artifact production of HMHP;30 on the other hand, at low pH, significant (even quantitative) interference by SO2 can occur whether or not
(42) Dasgupta, P. K.; Lindgren, P. F. Environ. Sci. Technol. 1989, 23, 895-897. (43) Dasgupta, P. K. ACS Adv. Chem. Ser. 1993, No. 232, 41-90.

HCHO is added.26,30 It is obviously difficult to use collector pH variation-based schemes26 to determine different peroxide species if significant concentrations of SO2 will be present. The Nafion membrane DS represents an interesting dilemma. Since the device is a diffusive collector and SO2 has a significantly smaller diffusion coefficient than H2O2, this is an advantage. Since Nafion is a cation-exchange membrane, it excludes S(IV) or other anions derived from it to pass through it and this may also be an advantage. However, if the exterior surface of the membrane is effectively wet and reaction can take place there, then the highly acidic nature of Nafion may actually catalyze the H2O2-S(IV) reaction. The actual interference study was conducted over several days. These results, shown in Table 1, suggest that the more favorable of the two possibilities listed above. As shown in Table 1, the experiments were conducted both at a constant level of H2O2 (1 ppbv) with variable levels of SO2 (0-200 ppbv) and at a constant level of SO2 (150 ppbv) with 1-5 ppbv levels of H2O2. The interference equivalent for the entire data set averages 1.43 ( 0.27 pptv H2O2 loss per ppbv SO2. Although this loss is not insignificant at low levels of H2O2 and high levels of SO2, this performance is far better than other collection methods; if a total collector was used, SO2 will interfere with H2O2 on a completely equivalent basis. Ozone. Ozone can produce H2O2 by surface-catalyzed reactions with water.27 Some degree of ozone interference is therefore to be expected. Ozone concentrations between 0 and 180 ppbv (n ) 5, at least six points at each concentration) were studied. The resulting data showed a linear interference by ozone (r2 ) 0.99) with an intercept statistically indistinguishable from zero. The ozone interference equivalent was determined to be

1 ppb ozone ) 2.37 ppt H2O2

(3)

All field data were corrected with this assumed degree of ozone interference based on simultaneously measured ozone levels. This may result in slight overcorrection in that when ozone interference was measured in the presence of 2.0 ppb H2O2 and 50-100 ppbv O3, the observed interference was less than what would be predicted by eq 3. In any case, the overall degree of interference

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is not very high and is generally ameliorated by the fact that concentrations of H2O2 are also typically higher during high ozone episodes. Methyl Hydroperoxide. With substrates other than thiamine it has been previously observed that hematin is significantly more selective for H2O2 than for alkylhydroperoxides.14 Potential interference from MHP was studied both in the solution phase and in the gas phase for the thiamine-hematin system. When a standardized MHP solution (20.3 and 40.6 M), from which H2O2 had been freshly removed by passing through a MnO2 column, was injected in the analytical system, the response was found to be 4.6 ( 0.1 and 5.3 ( 0.1%, respectively, of the same concentration of H2O2. Additive tests indicated an even lower degree of interference. For 2 M H2O2, the addition of 50.8 M MHP led to 5.5 ( 0.1% positive error in the determination. The same experiment with 4 M H2O2 showed that the addition of 101.6 M MHP resulted in a positive error of 9.5 ( 0.2%. The difference with the MHP only experiments is likely due to the preferential binding of H2O2 to hematin to produce the active intermediate. Such a process would leave the catalyst unavailable for MHP and lead to even lower interference than pure MHP experiments. For the gas phase, the best-fit linear regression equations for the response behavior were as follows over the ranges of 0-10.0 ppbv H2O2 and MHP individually (5 concentration points each, g3 measurements at each concentration):

Figure 6. Typical system output, gas-phase samples; concentration as indicated; 2-min sample, 8-min zero.

H2O2 (peak height, A/D count) ) 3102 ( 53 [H2O2, ppbv] - (148.5 ( 35.0), r2 ) 0.9991 (4) MHP (peak height, A/D count) ) 37.8 ( 1.3 [MHP, ppbv] + 44.4 ( 8.6, r2 ) 0.9967 (5)
The response of gas-phase MHP is thus only 1% of that of the same concentration of H2O2. This is due to the lower Henrys law solubility and diffusion coefficient of MHP relative to H2O2 and hence a lower collection efficiency (vide infra). Because of the very low potential interference from MHP, mixed gaseous standards of H2O2 and MHP, which for reasons outlined above are likely to lead to an even lower interference index for MHP, were not tested. If HRP is substituted for hematin in the reaction system, significant responses for MHP can be observed. The collection efficiency of the present DS (water as scrubber liquid) to MHP was assessed with a HRP (32 purpurogallin units/mL) -thiamine (1 mg/mL) reaction system with a reaction pH of 9.0 (phosphate buffer). Both DS units show essentially identical behavior, exhibiting a fractional collection efficiency of 0.318, 0.284, 0.275, 0.265, and 0.214 at flow rates of 0.76, 1.07, 1.36, 1.74, and 2.40 L/min, respectively. The fit of these data to a diffusion-controlled collection equation (e.g., eq 1) is very poor (r2 ) 0.83), and if a diffusion controlled collection regime is assumed nevertheless, the best-fit diffusion coefficient is calculated to be a factor of 15 less than that of H2O2; this is clearly impossible. The collection of MHP is therefore controlled by factors other than diffusive mass transport, notably the sink efficiency of the collection liquid and the solubility of MHP in water. It is possible that a different collection liquid

(e.g., containing an alcohol or a surfactant) may improve collection efficiency for MHP; however, compatibility with the DS and the analysis system must also be ensured. Hydroxymethyl Hydroperoxide. In all of the following, by HMHP, we mean a mixture that contains mostly HMHP. According to our analysis and the literature,37 our best estimate for the composition of this mixture, the molar ratio of BHMHP:HMHP:H2O2 is 1:7.5:2.6. In terms of its aqueous solution behavior, when a solution of HMHP (or BHMHP) is injected into a strongly alkaline aqueous medium like the present analysis system, it should hydrolyze to H2O2 very rapidly. When a crude HMHP mixture is freshly passed through an MnO2 bed and the resulting effluent is immediately analyzed by injection into the present analysis system and by iodometric analysis, the fluorescence results (treated on the basis of a H2O2 calibration plot) account for 92-94% of the total peroxides determined by iodometry. Given the difficulties of accurate calibrations of mixtures of different peroxides, we believe these results largely confirm the theoretical expectation that, in the solution phase, the system responds to any HMHP in the same way as H2O2. Regarding the behavior of gas-phase HMHP, in contrast to MHP, the dependence of HMHP collection efficiency upon flow rate does closely follow a diffusion-limited collection behavior (flow rate 0. 76-2.4 L/min, collection efficiency 88.5-55.7%):

ln(1 - f) ) -(1.4863 ( 0.0774)/Q - 0.2352 ( 0.0643, r2 ) 0.9946 (6)

In as much as HMHP displays higher solubility than H2O2 (the Henrys law coefficient of BHMHP is even higher4), it is reasonable to assume that, like HMHP, it too has very high Henrys law solubility), diffusion-limited collection behavior is expected. Further, the ratio of the slope in eq 4 to the mean slope in eqs 1 and 2 is 0.79. Nominally, this is the ratio of the diffusion coefficient of HMHP to H2O2. We can estimate the diffusion coefficient of HMHP on the basis of the weighted average of Grahams lawAnalytical Chemistry, Vol. 72, No. 21, November 1, 2000
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Figure 7. Hydrogen peroxide concentrations measured in Atlanta, GA, August 1999 In this and the subsequent figures, by hydrogen peroxide we mean the sum of H2O2 and HMHP.

based diffusion coefficients of BHMHP, HMHP, and H2O2. On the basis of the molecular weight of the individual compounds and the molar ratio of the different compounds in the mixture, the diffusion coefficient of HMHP is computed to be 0.80 relative to H2O2 being unity. This is obviously in excellent agreement with the experimental observations. Analytical Performance for Hydrogen Peroxide. For the liquid-phase system, the observed fluorescence signal was linear with H2O2 concentration in the tested range of 0-100 M:

peak height, A/D count ) 2696 ( 25 [H2O2, M] -9.2 ( 15.2, r2 ) 0.9996 (7)
Calibration linearity data for gas-phase H2O2 has already been presented in the previous section. A typical chart output for system response to 0-1ppbv H2O2(g) is shown in Figure 6. The limits of detection (S/N ) 3) for gaseous and aqueous H2O2 were computed to be 13.5 pptv and 11 nM, respectively. The relative standard deviation for the determination of 1 ppbv H2O2(g) and 1 M H2O2(aq) were 1.8 and 1.4%, respectively (n ) 7 each). Field Study Results. We cannot obviously differentiate between H2O2 and HMHP at this time. However, recent evidence based on chromatographic analysis suggests that H2O2 and MHP are the only two important atmospheric peroxides;24,44 as such, we cannot be in gross error in assuming what we measure to be H2O2. The H2O2 concentrations measured during the study ranged from less than 200 pptv (usually shortly after rainfall) to 4.5 ppbv (during a day when the ozone concentration exceeded 140 ppbv). A portion of the H2O2 results is shown in Figure 7. Detailed results will appear in the NARSTO database.45 The characteristic diurnal pattern for H2O2 is readily apparent in Figure 7.
(44) http://euros.gso.uri.edu/snow/instrumentschem.html. (45) http://www.cgenv.com/Narsto/.

Figure 8. Daily ozone maximum (hourly maximum measured among any of several metropolitan Atlanta measurement sites36) and daily H2O2 maximum measured at the Supersite experimental site.

Figure 8 shows a trace of maximum hourly ozone concentrations (taken among the several measurement sites located in metropolitan Atlanta)36 each day and the daily maximum in H2O2. There is obviously a striking similarity in these temporal patterns. A detailed rationale is beyond the scope of this paper. Figure 9 shows a shorter span vignette of H2O2, SO2, and O3 data, along with rainfall, during a mild rain episode. During August 8, the H2O2, SO2, and O3 all peak at the same time, suggesting a common plume source impacting the sampling location. Fossil fuel combustion sources often result in the quixotic combination of both elevated SO2 and oxidant levels; we have made similar observations in airborne monitoring of Ships plumes.46 Interestingly, rain washes out H2O2 and SO2, the two water-soluble gases, quite

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Figure 9. H2O2, ozone, SO2, and rainfall data during a 2-day period in Atlanta, GA, August 1999.

effectively, and ozone is removed to a much smaller degree. In fact, ozone concentrations go up with the second burst of showers, which was accompanied by significant lightning activity. On August 9, SO2 and ozone both reach higher levels than the previous day but H2O2 levels remain generally low and generally anticorrelated to SO2 concentrations, as may happen when the air mass has been processed at least partially through a cloud. During a total deployment period of 39,300 min, the instrument was collecting data 96.88% of the total time; calibration required 1.54% and reagent replacement/cleaning required 1.27% of the total time. The instrument was inoperative 0.31% of the total time, due to power failure resulting from rainwater running down the exterior of the sampling conduit and shorting out a power socket. The compact size of the instrument, largely due to the unique fluorescence detector, (entire instrument is built within a tower(46) Genfa, Z.; Dasgupta, P. K.; Frick, G. M.; Hoppel, W. A. Microchem. J. 199, 62, 99-113.

style PC chassis) makes for facile field deployment. The zero air self-purification design was also particularly useful in field applications in that it obviated the need for compressed gas cylinders altogether. It was found that the activated carbon column (1.5 cm 15 cm) still removes H2O2 quantitatively after more than 1 month of continuous use. Future Work. It is not our intention to suggest that this type of analysis will replace chromatographic analysis of peroxides. However, the present instruments are capable of simpler alternatives. If indeed it is sufficient to determine H2O2 and MHP, this can be accomplished by two parallel systems in which hematin and HRP are respectively used and lead to signals that are proportional to [H2O2] and [H2O2] + x[MHP], where x is of the order of 0.2. It may be possible to increase the value of x, by enhancing the collection of MHP through the incorporation of a modifier, e.g., a surfactant or an organic solvent in the scrubber liquid. If HMHP concentrations are important, then both systems will be measuring [H2O2] + y[HMHP] instead of [H2O2], where y is of the order of 0.8. In this case, a third channel can be operated with a iron phthalocyanine catalyst that can work in an acid solution15 and should thus provide a method of determining H2O2 individually without significant reaction of HMHP. However, the attractiveness of direct chromatographic analysis obviously increases as the number of components increase. ACKNOWLEDGMENT This research was funded in part by the SOS/Supersite Research Program of the USEPA. The manuscript was not subject to review by any of these agencies and no endorsement should be inferred. We are grateful to W. L. Chameides, S. V. Hering, and C. S. Kiang for asking us to be a part of this study. The help of E. Edgerton, Jefferson street marshall, before, during, and after the study, is gratefully acknowledged. Received for review May 31, 2000. Accepted August 28, 2000.
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