Determination of polycyclic aromatic hydrocarbons using lab on valve dispersive liquid-liquid microextraction coupled to high performance chromatography.

In this work, dispersive liquid-liquid microextraction (DLLME) method was applied for high performance liquid chromatography (HPLC) determination of 15 PAHs in aqueous matrices.The extraction procedure was automated using a system of multisyringe flow injection analysis coupled to HPLC instrument with fluorescence detector. Factors affecting the extraction process, such as type and volume of extraction and dispersive solvent, extraction time and centrifugation step were investigated thoroughly and optimized utilizing factorial design. The best recovery was achieved using 100 µL of trichloroethylene as the extraction solvent and 900 µL of acetonitrile as the dispersive solvent.The results showed that extraction time has no effect on the recovery of PAHs. The enrichment factors of PAHs were in the range of 86-95 with limits of detection of 0.02-0.6 µg L(-1). The linearity was 0.2-600 µg L(-1) for different PAHs. The relative standard deviation (RSD) for intra- and inter-day of extraction of PAHs were in the range of 1.6-4.7 and 2.1-5.3, respectively, for five measurements.The developed method was used to assess the occurrence of 15 PAHs in tap water, rain waters and river surface waters samples.

Multisyringe chromatography (MSC) using a monolithic column for the determination of sulphonated azo dyes.

A methodology based on multisyringe chromatography with a monolithic column was developed to determine three sulphonated azo textile dyes: Acid Yellow 23, Acid Yellow 9 and Acid Red 97. An ion pair reagent was needed because of the low affinity between the monolithic column and the anionic dyes. The proposed analytical system is simple, versatile and low-cost and has great flexibility in manifold configuration. The method was optimized through experimentation based on experimental design methodology. For this purpose two blocks of full factorial 2(3) were done sequentially. In the first experimental plan, the factors studied were: the % of acetonitrile in organic phase, the % of H(2)O in the mobile phase and the kind of ion pair reagent. In this stage, a simple configuration was used which has only one syringe for the mobile phase. After the first experimentation, we added a second syringe with a second mobile phase to the multisyringe module and performed a second full factorial 2(3). The factors studied in this case were: the % of acetonitrile in the second mobile phase, the pH and the concentration of ion pair reagent in both mobile phases. After this design, the optimal conditions were selected for obtaining a good resolution between the peaks of yellow dyes (1.47) and the elution of red dye in less than 8 min. The methodology was validated by spiking different amounts of each dye in real water samples, specifically, tap water, well water and water from a biological wastewater lagoon.

A multisyringe sequential injection method for monitoring water in the energy cogeneration system of a municipal waste incinerator.

Leading-edge urban solid waste ashing plants use burning heat energy to obtain electrical power. Water fed to their boilers for conversion into steam should be highly pure in order to minimize corrosion, scaling and similar phenomena, which can lead to malfunctioning and a reduced useful life but can be avoided by proper management and control of the water supply. In this work, we developed a multiparameter monitor based on multisyringe sequential injection for the sequential determination of up to eight important parameters, namely: pH, specific and acid conductivity, hydrazine, ammonium, phosphate, silicate and total iron. Acid conductivity was determined by passing the sample through a cation-exchange resin in order to retain ammonium ion and release protons. This parameter was deemed the most accurate indicator of dissolved solids in boiler water. Chemical parameters were determined spectrophotometrically: hydrazine by reaction with p-dimethylaminobenzaldehyde, ammonium by the modified Berthelot reaction, iron with o-phenanthroline, and phosphate and silica by formation of a molybdoheteropoly blue dye in the presence of ascorbic acid as reductant. Use of the optimum chemical and physical operating conditions provided 3s(blank) detection limits of 0.01 mg l(-1) N(2)H(4), 0.13 mg l(-1) NH(4)(+), 0.04 mg l(-1) Fe, 0.03 mg l(-1) SiO2 and 0.05 mg l(-1) PO(4)(3-), and relative standard deviations not greater than 2.5%. The methods integrated in the proposed monitor were successfully applied to real samples from the water-steam cycle at the Son Reus ashing plant in Palma de Mallorca (Spain).

Potential of multisyringe flow-based multicommutated systems.

This work is aimed at emphasizing the potential of the multicommutated systems based on the multisyringe flow injection analysis (MSFIA) modality. First, the characteristics, advantages and withdraws offered by flow analysis systems based on the different non-segmented modalities are briefly described. In these systems, multicommutation and computer control of the analytical process occupy a predominant place, as in the case of sequential injection analysis (SIA), multicommutated flow injection analysis (MCFIA), MSFIA and multipumping flow systems (MPFS). Next, several examples are given and different aspects of the implementation of analysers based on MSFIA designs for the construction of different analysis systems, including intelligent (smart) systems, use of sample pre-treatment automatic systems, for chromatographic and non-chromatographic determinations as well as use of monolithic or capillary electrophoresis columns are considered.

Determination of mercury by multisyringe flow injection system with cold-vapor atomic absorption spectrometry.

A new software-controlled time-based multisyringe flow injection system for mercury determination by cold-vapor atomic absorption spectrometry is proposed. Precise known volumes of sample, reducing agent (1.1% SnCl2 in 3% HCl) and carrier (3% HCl) are dispensed into a gas-liquid separation cell with a multisyringe burette coupled with one three-way solenoid valve. An argon flow delivers the reduced mercury to the spectrometer. The optimization of the system was carried out testing reaction coils and gas-liquid separators of different design as well as changing parameters, such as sample and reagents volumes, reagent concentrations and carrier gas flow rate, among others. The analytical curves were obtained within the range 50-5000 ng L(-1). The detection limit (3sigma(b)/S) achieved is 5 ng L(-1). The relative standard deviation (R.S.D.) was 1.4%, evaluated from 16 successive injections of 250 ng L(-1) Hg standard solution. The injection and sample throughput per hour were 44 and 11, respectively. This technique was validated by means of solid and water reference materials with good agreement with the certified values and was successfully applied to fish samples.

Speciation analysis of inorganic arsenic by a multisyringe flow injection system with hydride generation-atomic fluorescence spectrometric detection.

In this study, a new technique by hydride generation-atomic fluorescence spectrometry (HG-AFS) for determination and speciation of inorganic arsenic using multisyringe flow injection analysis (MSFIA) is reported. The hydride (arsine) was generated by injecting precise known volumes of sample, a reducing sodium tetrahydroborate solution (0.2%), hydrochloric acid (6M) and a pre-reducing solution (potassium iodide 10% and ascorbic acid 0.2%) to the system using a multisyringe burette coupled with one multi-port selection valve. This solution is used to pre-reduce As(V) to As(III), when the task is to speciate As(III) and As(V). As(V) is determined by the difference between total inorganic arsenic and As(III). The reagents are dispensed into a gas-liquid separation cell. An argon flow delivers the arsine into the flame of an atomic fluorescence spectrometer. A hydrogen flow has been used to support the flame. Nitrogen has been employed as a drier gas (Fig. 1). Several variables such as sample and reagents volumes, flow rates and reagent concentrations were investigated in detail. A linear calibration graph was obtained for arsenic determination between 0.1 and 3 microg l(-1). The detection limit of the proposed technique (3sigma(b)/S) was 0.05 microg l(-1). The relative standard deviation (R.S.D.) of As at 1 microg l(-1) was 4.4 % (n=15). A sample throughput of 10 samples per hour was achieved. This technique was validated by means of reference solid and water materials with good agreement with the certified values. Satisfactory results for speciation of As(III) and As(V) by means of the developed technique were obtained.

Characterization of bottom ash in municipal solid waste incinerators for its use in road base.

Incineration of municipal solid wastes (MSWs) produces by-products which can be broadly classified as bottom and fly ashes. Since MSW incineration started, possibilities other than landfilling the incineration residues have been sought; most initiatives in this sense tend to use these residues as aggregate substitute in pavements and other road construction elements. The main goal of the present work is the physical and chemical characterization of the local incineration bottom ash towards its eventual re-utilization. The study includes not only the specific aspects regarding its role as pavement element, but also the assessment of the environmental effects. Therefore, together with the determination of physical (moisture content, apparent and bulk densities, crystallinity, etc.) and engineering properties (particle size distribution, abrasion and impact resistance, etc.), full chemical characterization of the bottom ash and the study of leaching as a function of aging time have been undertaken. The results obtained indicate that the metal content of both the raw bottom ash and its leachates fulfill the environmental regulations provided that the bottom ash is stored for at least one month. Engineering properties of the bottom ash are close to those of natural aggregates and, thus, road-construction use of these residues seems to be feasible.

Preconcentration and determination of inorganic arsenic using a multisyringe flow injection system and hydride generation-atomic fluorescence spectrometry.

A new multisyringe flow injection system for inorganic arsenic determination at trace levels by hydride generation-atomic fluorescence spectrometry (HGAFS) is presented. Preconcentration on a solid-phase was carried out using a column packed with an anion-exchange resin (Amberlite IRA-410). The reagents are dispensed to the system using a multisyringe burette coupled with two multi-port selection valves. Different parameters were changing in order to make the system as effective as possible. An analytical curve was obtained for arsenic determination between 50 and 2000ngl(-1). This new approach improved five times the sensitivity over a MSFIA-HGAFS technique developed previously by the authors. Detection limit of the proposed technique was (3sigma(b)/S) of 30ngl(-1). The relative standard deviation (R.S.D.) of As at 1mugl(-1) was 4.8% (n=7). A sample throughput of 10h(-1) has been achieved. The proposed method has been applied to different reference solid and water materials with satisfactory results.

Elevated tissue kallikrein activity in airway secretions from patients with tracheobronchitis associated with prolonged mechanical ventilation.

The clinical course of patients undergoing prolonged mechanical ventilation is often complicated by the development of purulent tracheobronchitis. The purpose of this study was to assess whether ventilator-associated hypersecretion is associated with elevated levels of tissue kallikrein (TK) activity. TK can induce marked bronchial inflammation in animal models and TK activity is increased in the airway secretions of symptomatic asthmatics. It has not been studied in conditions with predominantly neutrophilic bronchial secretions, although animal data indicate that neutrophil elastase may stimulate TK activity. We measured TK activity in airway secretions of patients undergoing mechanical ventilation for more than 4 weeks (PMV group) and in two comparator groups: patients with cystic fibrosis, who were colonized with Pseudomonas aeruginosa (CF group) and patients undergoing mechanical ventilation for less than one week who did not have clinical evidence of purulent airway secretions (acute mechanical ventilation, AMV group). We also compared the level of neutrophil elastase (NE) activity, an index of neutrophil activation, in the three patient groups. TK and NE activity in the sol phase were measured by the degradation of chromogenic substrates (DL Val-Leu-Arg pNA and N-Methoxy Succinyl Ala-Ala-Pro-Val pNA, respectively). Intergroup differences in cell counts were not significant. However, TK activity was significantly less in the AMV group than in the PMV and cystic fibrosis patients (Kruskal-Wallis ANOVA, p < 0.05). Elastase activity was significantly greater in the CF group (p < 0.05) than in the other two groups. Compared to patients undergoing short-term mechanical ventilation (AMV group), TK activity was elevated in patients with purulent tracheobronchitis associated with prolonged mechanical ventilation (PMV group). The elevation in TK activity in these patients is comparable to levels in sputum from patients with cystic fibrosis (CF group), although the latter had a significantly higher level of NE activity. The observation of increased TK activity in patients with neutrophilic airway inflammation suggests that TK may play a role in modulating inflammation in ventilator-associated tracheobronchitis and may be worthy of further study to determine its source and significance.

Hyaluronic acid blocks porcine pancreatic elastase (PPE)-induced bronchoconstriction in sheep.

We previously showed that inhaled porcine pancreatic elastase (PPE) causes bronchoconstriction in sheep via a bradykinin-mediated mechanism. Hyaluronic acid (HA), in vitro, binds and inactivates airway tissue kallikrein (TK), the enzyme responsible for kinin generation. Therefore, we hypothesized that in vivo, HA should prevent PPE-induced bronchoconstriction by binding and inactivating TK. To test this, we measured pulmonary resistance (RL) in allergic sheep before and after inhalation of PPE alone (500 microg) and after pretreatment with either inhaled HA at 70 kD, designated low molecular weight (LMW)-HA or 200 kD, designated high molecular weight (HMW)-HA at different concentrations. Inhaled PPE increased RL 147 +/- 8% over baseline values and this effect was associated with a 111 +/- 28% increase in bronchoalveolar lavage fluid (BALF) TK activity. HA blocked the PPE-induced bronchoconstriction and the increase in BALF TK activity in a dose- dependent and molecular weight-dependent fashion. HA alone had no effect on RL. Instillation of PPE in the lung increased kinin concentrations in BALF, a result consistent with the PPE-induced increase in BALF TK activity. Our findings show that HA blocks PPE-induced bronchoconstriction in a dose-dependent and molecular weight-dependent fashion by a mechanism that may, in part, be related to inhibition of TK activity and the formation of kinins.

Hyaluronan serves a novel role in airway mucosal host defense.

Enzymes secreted onto epithelial surfaces play a vital role in innate mucosal defense, but are believed to be steadily removed from the surface by mechanical actions. Thus, the amount and availability of enzymes on the surface are thought to be maintained by secretion. In contrast to this paradigm, we show here that enzymes are retained at the apical surface of the airway epithelium by binding to surface-associated hyaluronan, providing an apical enzyme pool 'ready for use' and protected from ciliary clearance. We have studied lactoperoxidase, which prevents bacterial colonization of the airway, and kallikrein, which mediates allergic bronchoconstriction that limits the inhalation of noxious substances. Binding to hyaluronan inhibits kallikrein, which is needed only in certain situations, whereas lactoperoxidase, useful at all times, does not change its activity. Hyaluronan itself interacts withthe receptor for hyaluronic acid-mediated motility (RHAMM or CD168) that is expressed at the apex of ciliated airway epithelial cells. Functionally, hyaluronan binding to RHAMM stimulates ciliary beating. Thus, hyaluronan plays a previously unrecognized pivotal role in mucosal host defense by stimulating ciliary clearance of foreign material while simultaneously retaining enzymes important for homeostasis at the apical surface so that they cannot be removed by ciliary action.

Fast sequential injection determination of benzo[A]pyrene using variable angle fluorescence with on-line solid-phase extraction.

A methodology for the analysis of drinking water for one of the most potent carcinogenic agents known; benzo[a]pyrene (BaP), in the presence of other interfering PAHs is presented. The methodology described is based on the sequential injection analysis of the sample on to a microcolumn (containing 5 mg of C18) where extraction and preconcentration of BaP takes place, followed by elution of BaP with 1 ml of 1,4-dioxane and subsequent detection by using variable angle fluorescence. The advantages of the method include the small amount of stationary phase employed together with the possibility of re-using the phase in order to carry out a large number of injections without the need for column re-packing. Also noteworthy is the small volume of 1,4-dioxane used to elute the BaP retained on the column and the small sample volumes required (9-10 ml) for achieving detection limits at the ng l-1 level. Thus, a methodology for BaP determination is obtained which complies with the requirements of the 98/83/EC Directive which fixes a maximum admissible concentration for this pollutant in waters for public consumption of 10 ng l-1. The variable angle spectra obtained are further processed by means of the multiple linear regression technique. The detection limit for BaP is 2.5 ng l-1, and the linear range is between 7.5 and 280 ng l-1.

Sequential-injection procedure for determination of iodide in pharmaceutical and drinking water samples by catalytic reaction with spectrophotometric detection.

A novel sequential-injection system was developed for determination of iodide at very low concentrations by using a kinetic method. The method is based on the catalytic effect of iodide on the redox reaction between Ce4+ and As3+ first described by Sandell and Kolthoff. The calibration curve is constructed by measuring the decrease of Ce4+ absorbance versus iodide concentration with a delay time of 30 s. The detection limit is 1.5 microg/L, the working temperature is 45 degrees C, and the working range is 0.002-0.5 mg/L. Reasonable agreement was obtained when the method was applied to pharmaceutical and drinking water samples. The method has a sample throughput of approximately 15/h.

Secretory leukocyte protease inhibitor, but not alpha-1 protease inhibitor, blocks tryptase-induced bronchoconstriction.

Alpha-1-protease inhibitor (alpha(1)-PI) and secretory leukocyte protease inhibitor (SLPI) are two natural airway serine protease inhibitors. While inhibition of neutrophil elastase is a function common to both alpha(1)-PI and SLPI, we showed previously that they exhibit different patterns of protection against antigen-induced changes in airway function in allergic sheep. Specifically, the protective effect seen with SLPI was similar to the profile of action of synthetic tryptase inhibitors in the model. Based on these data, and the fact that tryptase is a serine protease, we hypothesized that SLPI, but not alpha(1)-PI, would block tryptase-induced bronchoconstriction. To test this, we compared the responses to inhaled tryptase in five sheep without treatment or after treatment with either aerosol alpha(1)-PI (10 mg) or aerosol SLPI (50 mg). The doses of alpha(1)-PI and SLPI selected had been shown to be effective in previous antigen-provocation studies. Treatments were given 30 min before aerosol challenge with tryptase (500 ng). Tryptase alone increased (mean+/-SEM) pulmonary resistance (R(L)) 142 +/- 24% over baseline. Pretreatment with alpha(1)-PI had no effect on the tryptase response (R(L)increased 122 +/- 20%). Pretreatment with SLPI, however, blocked the tryptase-induced response (R(L) increased only 40 +/- 4% P<0.05 vs. tryptase). These are the first studies comparing the inhibitory activity of SLPI and alpha(1)-PI on inhaled tryptase-induced bronchoconstriction. We conclude that, in vivo, SLPI, but not alpha(1)-PI, can block tryptase-induced bronchoconstriction and that this activity may explain the differential effects of these two serine protease inhibitors on antigen-induced airway responses in allergic sheep.

Hyaluronic acid in cultured ovine tracheal cells and its effect on ciliary beat frequency in vitro.

Hyaluronic acid (hyaluronan, or HA) is secreted by submucosal glands, but its function in airway secretions other than influencing the rheology of mucus is not fully understood. HA is known to modulate cell behavior and to enhance sperm motility. Because sperm tails and cilia have the same microtubular structure, we studied the effect of HA on ciliary beat frequency (CBF) in vitro. CBF of cultured ovine airway epithelial cells was measured continuously by digital video microscopy. After removal of endogenous HA by hyaluronidase, cells were exposed to 50 to 100 microg/mL of HA at different times in culture. No change in CBF in response to HA was seen in cells cultured less than 7 days. After 7 days, however, 6 of 10 measured cells (from three different sheep) showed a transient CBF increase from a baseline of 6.4 +/- 0.3 Hz (mean +/- SE) to 7.4 +/- 0.4 Hz or 16% above baseline (p < 0.05). At these time points (but not before), cytochemical staining was positive for endogenous HA using a biotinylated HA-binding protein. These data suggest that HA can increase CBF of tracheal epithelial cells only late in culture when HA is able to bind to an unspecified cell surface structure. Because this binding has a physiological effect, we hypothesize that it is an HA-binding receptor, that is either transiently expressed late in culture or initially destroyed by the protease treatment for cell dispersion.

Inhaled porcine pancreatic elastase causes bronchoconstriction via a bradykinin-mediated mechanism.

Neutrophil elastase has been linked to inflammatory lung diseases such as chronic obstructive pulmonary disease, adult respiratory distress syndrome, emphysema, and cystic fibrosis. In guinea pigs, aerosol challenge with human neutrophil elastase causes bronchoconstriction, but the mechanism by which this occurs is not completely understood. Our laboratory previously showed that human neutrophil elastase releases tissue kallikrein (TK) from cultured tracheal gland cells. TK has been identified as the major kininogenase of the airway and cleaves both high- and low-molecular weight kininogen to yield lysyl-bradykinin. Because inhaled bradykinin causes bronchoconstriction and airway hyperresponsiveness in asthmatic patients and allergic sheep, we hypothesized that elastase-induced bronchoconstriction could be mediated by bradykinin. To test this hypothesis, we measured lung resistance (RL) in sheep before and after inhalation of porcine pancreatic elastase (PPE) alone and after pretreatment with a bradykinin B(2) antagonist (NPC-567), the specific human elastase inhibitor ICI 200,355, the histamine H(1)-antagonist diphenhydramine hydrochloride, the cysteinyl leukotriene 1 receptor antagonist montelukast, or the cyclooxygenase inhibitor indomethacin. Inhaled PPE (125-1,000 microg) caused a dose-dependent increase in RL. Aerosol challenge with a single 500 microg dose of PPE increased RL by 132 +/- 8% over baseline. This response was blocked by pretreatment with NPC-567 and ICI-200,355 (n = 6; P < 0.001), whereas treatment with diphenhydramine hydrochloride, montelukast, or indomethacin failed to block the PPE-induced bronchoconstriction. Consistent with pharmacological data, TK activity in bronchial lavage fluid increased 134 +/- 57% over baseline (n = 5; P < 0.02). We conclude that, in sheep, PPE-induced bronchoconstriction is in part mediated by the generation of bradykinin. Our findings suggest that elastase-kinin interactions may contribute to changes in bronchial tone during inflammatory diseases of the airways.

The lactoperoxidase system functions in bacterial clearance of airways.

Airway mucus is a complex mixture of secretory products that provides a multifaceted defense against pulmonary infection. Mucus contains antimicrobial peptides (e.g., defensins) and enzymes (e.g., lysozyme) although the contribution of these to airway sterility has not been tested in vivo. We have previously shown that an enzymatically active, heme-containing peroxidase comprises 1% of the soluble protein in sheep airway secretions, and it has been hypothesized that this airway peroxidase may function as a biocidal system. In this study, we show that sheep airway peroxidase is identical to milk lactoperoxidase (LPO) and that sheep airway secretions contain thiocyanate (SCN(-)) at concentrations necessary and sufficient for a functional peroxidase system that can protect against infection. We also show that airway LPO, like milk LPO, produces the biocidal compound hypothiocyanite (OSCN(-)) in vitro. Finally, we show that in vivo inhibition of airway LPO in sheep leads to a significant decrease in bacterial clearance from the airways. The data suggest that the LPO system is a major contributor to airway defenses. This discovery may have significant implications for chronic airway colonization seen in respiratory diseases such as cystic fibrosis.

Bronchial tissue kallikrein activity is regulated by hyaluronic acid binding.

Tissue kallikrein (TK) is secreted by serous cells of tracheobronchial submucosal glands and plays a role in allergic airway responses. To better understand the regulation of TK, we used primary cultures of submucosal gland cells that release TK upon stimulation. Media from cultures stimulated with chymase (10(-7) M) showed increased TK activity (0.50 +/- 0.22 mU/ml mean +/- standard error) in comparison with the control group (0.08 +/- 0.02 mU/ml). The increased TK activity was significantly correlated with increases in the levels of the serous cell marker, secretory leukoprotease inhibitor. Anion exchange chromatography of the conditioned culture media showed that TK activity eluted as a broad peak between 1.6 and 1.8 M NaCl, unlike the reported elution (0.3 to 0.6 M NaCl) of kallikreins from other tissues, suggesting that secreted bronchial TK was bound to a negatively charged molecule. Hyaluronidase digestion increased TK activity in both pre- and post-chymase-stimulated culture media, whereas no such change was seen after samples were digested with heparinase or chondroitinase ABC. Further, after hyaluronidase digestion of media, TK eluted from an anion exchange column between 0.3 and 0.6 M NaCl. Enzymatic detection of TK after nondenaturing gel electrophoresis showed that hyaluronidase digestion also reduced the electrophoretic heterogeneity of TK to a single band, whereas adding back hyaluronic acid (HA) to hyaluronidase-digested samples restored the original heterogeneity. Finally, TK activity bound to HA-Sepharose and could be eluted with HA. These studies show that primary cultures of ovine submucosal gland cells secrete TK in a regulated fashion, and that secreted TK binds to HA. This binding reduces TK enzymatic activity; therefore, factors that affect HA turnover could modify the TK activity in the airway lumen. These events could be important in the regulation of kinin-mediated airway inflammation.

Flow techniques in water analysis.

In the present work the main flow techniques for the analysis and monitoring of several parameters of interest in the quality control of different types of waters are reviewed. Firstly, a review involving the advantages and disadvantages of flow techniques, from those currently out-dated, such as segmented flow analysis (SFA), to the most modern techniques, such as flow injection analysis (FIA), sequential injection analysis (SIA) and multi-commutation techniques (MCFA), is carried out. On the other hand, a new technique, the multi-syringe flow analysis (MSFA) is hereby described for the first time as both a fast and robust alternative. Its possibilities, limitations and potential advantages when using this technique either on its own or coupled to SIA, which carries out a previous sample handling, are outlined.

Isolation and characterization of a peroxidase from the airway.

Sheep airway mucus can potently scavenge hydrogen peroxide, an important mediator of airway inflammation. Here, the scavenging activity was identified as a peroxidase produced by goblet cells of the airway epithelium and secreted into the airway lumen. Ovine airway peroxidase activity was purified approximately 100-fold from airway lavage fluid in two steps, using cation exchange and lectin affinity chromatography, yielding an apparently homogeneous 82-kD glycoprotein. Ovine airway peroxidase represented about 1% of the total protein in airway mucus and thus was an abundant enzyme in airway secretions. The absorbance spectrum of the purified peroxidase showed a major peak at 412 nm indicative of a hemoprotein. The ratio of A412/A280 of the purified enzyme was 0.86. The absorption spectrum of ovine airway peroxidase, its ability to oxidize halides, its sensitivity to inhibitors and its apparent molecular mass on sodium dodecyl sulfate gels showed that airway peroxidase was similar to lactoperoxidase but distinguished from myeloperoxidase, eosinophil peroxidase as well as from glutathione peroxidases. Based on these observations, ovine airway peroxidase is a newly isolated and abundant enzyme of airway mucus which may function to control reactive oxygen species in the airway and to prevent infection by catalyzing the formation of biocidal compounds.