Half-life In Air Research Articles

Determination of environmental properties and toxicity of octyl-dimethyl-para-aminobenzoic acid and its degradation products

Octyl-dimethyl-para-aminobenzoic acid (ODPABA) is one of compounds of emerging concern. It undergoes transformations under the influence of oxidizing or chlorinating agents and UV radiation forming products with different properties. There is very little experimental data concerning the environmental fate of ODPABA and its transformation products. Therefore, the purpose of the studies was to determine environmental parameters: water solubility, soil – water partition coefficient, octanol – air partition coefficient, bioconcentration factor as well as half-life in air, water and soil. Based on the results obtained, the persistence and migration possibilities of ODPABA and its transformation products in the aquatic environment were estimated. Moreover, the ecological toxicity of oxidation and chlorination products was investigated. Microtox®, Daphtoxkit F® and Artoxkit M® tests were used to determine toxicity. LC50 for Fish and Daphnia magna was calculated by Ecosar module. Studies have shown that as a result of ODPABA transformations, chloroorganic products are formed, which are lipophilic, are bioconcentrated in organic matter, are characterized by significant environmental persistence, can spread over considerable distances and are toxic. Oxidation products have significantly smaller impact on the environment. They are characterized by higher water solubility, lower bioconcentration factor and are less toxic.

Passive Air Samplers As a Tool for Assessing Long-Term Trends in Atmospheric Concentrations of Semivolatile Organic Compounds

Many attempts have been made to quantify the relationship between the amount of persistent organic pollutants sequestered by passive air sampling devices and their actual concentrations in ambient air. However, this information may not be necessary for some applications. In this study, two sets of 30 ten-year-long time series of simultaneous passive and high-volume active air sampling carried out at the Košetice observatory in the Czech Republic were used for a comparison of temporal trends. Fifteen polyaromatic hydrocarbons, seven polychlorinated biphenyls and eight organochlorine pesticides were investigated. In most cases, a good agreement was observed between the trends derived from passive and active monitoring with the exception of several compounds obviously affected by sampling artifacts. Two sampling artifacts were observed: breakthrough of high-volume sampler filters for penta- and hexachlorobenzene and semiquantitative values for PAHs with a high molecular weight. It has been suggested before that annually aggregated results of passive air monitoring may be used directly for the assessment of the long-term behavior of these compounds. The extensive set of long-term data used in this study allowed us to confirm this finding and to demonstrate that it is also possible to derive temporal trends and the compounds' half-lives in air from the passive-sampling time series.

Remoteness from sources of persistent organic pollutants in the multi-media global environment

Quantifying the remoteness from sources of persistent organic pollutants (POPs) can inform the design of monitoring studies and the interpretation of measurement data. Previous work on quantifying remoteness has not explicitly considered partitioning between the gas phase and aerosols, and between the atmosphere and the Earth's surface. The objective of this study is to present a metric of remoteness for POPs transported through the atmosphere calculated with a global multimedia fate model, BETR-Research. We calculated the remoteness of regions covering the entire globe from emission sources distributed according to light emissions, and taking into account the multimedia partitioning properties of chemicals and using averaged global climate data. Remoteness for hypothetical chemicals with distinct partitioning properties (volatile, semi-volatile, hydrophilic, low-volatility) and having two different half-lives in air (60-day and 2-day) are presented. Differences in remoteness distribution among the hypothetical chemicals are most pronounced in scenarios assuming 60-day half-life in air. In scenarios with a 2-day half-life in air, degradation dominates over wet and dry deposition processes as a pathway for atmospheric removal of all chemicals except the low-volatility chemical. The remoteness distribution of the low-volatility chemical is strongly dependent on assumptions about degradability on atmospheric aerosols. Calculations that considered seasonal variability in temperature, hydroxyl radical concentrations in the atmosphere and global atmospheric and oceanic circulation patterns indicate that variability in hydroxyl radical concentrations largely determines the seasonal variability of remoteness. Concentrations of polybrominated diphenyl ethers (PBDEs) measured in tree bark from around the world are more highly correlated with remoteness calculated using our methods than with proximity to human population, and we see considerable potential to apply remoteness calculations for interpretation of monitoring data collected under programs such as the Stockholm Convention Global Monitoring Plan.

A modeling assessment of the physicochemical properties and environmental fate of emerging and novel per- and polyfluoroalkyl substances

Long-chain perfluoroalkyl carboxylic acids (PFCAs) and perfluoroalkane sulfonic acids (PFSAs) are persistent, bioaccumulative, and toxic contaminants that are globally present in the environment, wildlife and humans. Phase-out actions and use restrictions to reduce the environmental release of long-chain PFCAs, PFSAs and their precursors have been taken since 2000. In particular, long-chain poly- and perfluoroalkyl substances (PFASs) are being replaced with shorter-chain homologues or other fluorinated or non-fluorinated alternatives. A key question is: are these alternatives, particularly the structurally similar fluorinated alternatives, less hazardous to humans and the environment than the substances they replace? Several fluorinated alternatives including perfluoroether carboxylic acids (PFECAs) and perfluoroether sulfonic acids (PFESAs) have been recently identified. However, the scarcity of experimental data prevents hazard and risk assessments for these substances. In this study, we use state-of-the-art in silico tools to estimate key properties of these newly identified fluorinated alternatives. [i] COSMOtherm and SPARC are used to estimate physicochemical properties. The US EPA EPISuite software package is used to predict degradation half-lives in air, water and soil. [ii] In combination with estimated chemical properties, a fugacity-based multimedia mass-balance unit-world model – the OECD Overall Persistence (POV) and Long-Range Transport Potential (LRTP) Screening Tool – is used to assess the likely environmental fate of these alternatives. Even though the fluorinated alternatives contain some structural differences, their physicochemical properties are not significantly different from those of their predecessors. Furthermore, most of the alternatives are estimated to be similarly persistent and mobile in the environment as the long-chain PFASs. The models therefore predict that the fluorinated alternatives will become globally distributed in the environment similar to their predecessors. Although such in silico methods are coupled with uncertainties, this preliminary assessment provides enough cause for concern to warrant experimental work to better determine the properties of these fluorinated alternatives.

Current-Use Herbicides in Air as Influenced by Their Estimated Agricultural Use at Various Distances from Six Sampling Locations

Passive air samplers were deployed at six locations across the province of Manitoba, Canada, to represent areas with, or at various distances from, agricultural herbicide applications. During the growing seasons in 2008 and 2009, the four southern sites always demonstrated mixtures of current-use herbicides (CUHs) in air, but CUHs were not detected at the two northern sites that were 400 and 800 km away from Manitoba’s most northern boundary of agricultural herbicide applications. The masses of the CUHs detected in the air were most strongly positively associated with their estimated masses typically applied in a ~100-km2 township area surrounding the sampling sites (r = 0.70 to 0.74) and to a lesser extent with their estimated mass applied in incrementally larger areas (r = 0.53 to 0.59). The masses of CUHs detected in air were also significantly positively associated with their estimated masses applied at a provincial level (r = 0.45 to 0.52) but not with their reported half-lives in air, suggesting that a system of maintaining records of herbicide use data, even at a coarse scale, can strongly improve agri-environmental risk assessments. Of the nine CUHs detected, MCPA [(4-chloro-2-methylphenoxy)acetic acid] and bromoxynil, which are widely applied in Manitoba agriculture, were the only herbicides detected at all four southern sites. Triallate and metolachlor which have low use in Manitoba were the only CUHs detected in the winter months, confirming that these herbicides are relatively persistent in the air and may undergo long-range transport. Four passive air samplers, each installed 0.5 to 1.5 km apart at the same location, showed variations in the herbicide masses detected with the coefficient of variation ranging from 10 % for bromoxynil in 2008 to 137 % for MCPA in 2009. These variations were particularly observed not only for the two herbicides applied on-site (MCPA and clopyralid) but also for four herbicides transported into the area from longer distances (2,4-D (2,4-dichlorophenoxyacetic acid), bromoxynil, ethalfluralin, and triallate). Future regional-scale research should therefore consider deploying multiple sets of passive air samplers at a site to obtain a more representative measure of herbicide air concentrations.

Response to comment on “Identification of substances with potential for long-range transport as possible substances of very high concern”

In his comment on our article “Identification of substances with potential for long-range transport as possible substances of very high concern” [1], Sierra Rayne identified several substances, which might lead to another classification as indicated by us. We agree with Dr. Rayne that one substance ethanol, 2,2′,2′′-nitrilotris-, compound with (chlorometyl)oxirane (1:1) (CAS No. 68141-08-02) is significantly ionized in most aquatic systems. He further claimed that some compounds have been classified as persistent although they are rapidly hydrolyzed. We clearly described in the methods section of our article that we used the fugacity module of the EPI Suite for estimating degradation half-lives in air, water, sediment, and soil (p. 3154). The latter three half-lives are determined using the BIOWIN estimation program. “If a compound is susceptible to abiotic hydrolysis under environmental conditions, ... then the user must manually enter the value into the Fugacity, Half-life Selection option.” (EPI Suite, fugacity help file). We developed an automatic screening procedure for a large substance list without manually entered values and evaluated “the screening results for the identified 11 compounds...to underline that estimated parameters of the technically identified substances can be confirmed...” (p. 3159). We thank Dr. Rayne for checking the substances on their hydrolytic degradation potential, which is very helpful to prioritizing chemicals as possible substances of very high concern.

Comparaison de l’aérosol de la cigarette électronique à celui des cigarettes ordinaires et de la chicha

In previous studies of the smoke from regular cigarettes and water pipes, we measured aerosol particle sizes in three streams; S1, inhaled by the smoker, S2, released by the device itself and S3, exhaled by the smoker. We used an electrostatic low-pressure impactor (ELPI), giving particle size distributions in real time and calculated median diameters, D50, and dispersion (σg). This allowed us to predict airway deposition. In addition, the aerosol particle half-life in the air was used as a measure of the risk to others from passive smoking. With the same equipment, we measured the particle sizes and persistence in air of the liquid aerosol generated by e-cigarettes (Cigarettec®) containing water, propylene glycol and flavorings with or without nicotine. Aerosol generation was triggered by a syringe or by the inspiration of volunteer smokers. The D50 data obtained in S1, were 0.65 μm with nicotine and 0.60 μm without nicotine. Deposition in the airways could then be calculated: 26% of the total would deposit, of which 14% would reach the alveoli. These data are close to those found with regular cigarettes. For S3, D50 data were 0.34 μm and 0.29 μm with or without nicotine. The half-life in air of the S3 stream was 11 seconds due to a rapid evaporation. The-e-cigarette aerosol, as measured here, is made of particles bigger than those of cigarette and water pipe aerosols. Their deposition in the lung depends on their fate in the airways, which is unknown. Contrary to tobacco smoke, which has a half-life in air of 19 to 20 minutes, the risk of passive "smoking" exposure from e-cigarettes is modest.

Uncertainty analysis using a fugacity-based multimedia mass-balance model: Application of the updated EQC model to decamethylcyclopentasiloxane (D5)

The EQuilibrium Criterion (EQC) model developed and published in 1996 was recently revised to include improved treatment of input partitioning and reactivity data, temperature dependence and an easier sensitivity and uncertainty analysis. This New EQC model was used to evaluate the multimedia, fugacity-based fate of decamethylcyclopentasiloxane (D5; CAS No. 541-02-6) in the environment over a temperature range of 1–25°C. In addition, Monte Carlo uncertainty analysis was used to quantitatively determine the influence of temperature and input partitioning and reactivity data on the behavior of D5 under various emission scenarios. Results indicated that emission mode was the most influential factor determining the fate and distribution of D5 in the model environment. When emitted to air and soil, D5 partitioned to and remained in the air compartment where rates of removal from degradation and advection processes were relatively rapid. In contrast, D5 emitted to water resulted in a substantial mass fraction of D5 being accumulated in the sediment compartment, where rates of removal from degradation and advection processes were slow. The mass distributions and fate of D5 in the model environment were strongly influenced by multiple input parameters, including temperature, the mode of emission (especially emission rate to water), KOC and half-life in air. As temperature decreased from 25°C to 1°C, KOC and half-life in air became increasingly more influential such that the mass distribution of D5 increased in air and decreased in sediment, resulting in decreased overall persistence.

Screening Criteria for Long-Range Transport Potential of Organic Substances in Water

Screening of long-range transport potential (LRTP) of organic chemicals in water requires the development of criteria in analogy to the existing LRTP criteria for airborne chemicals. According to the Stockholm Convention, compounds mainly partitioning into air are assumed to be prone to LRTP if they have a half-life in air of more than two days. Using mean flow velocities of European rivers (0.7-1 m/s) and of ocean currents running into the Arctic Ocean (0.28-0.9 m/s), we derived corresponding half-life criteria for freshwater and seawater (10 days and 90 days, respectively). Next, we calculated the characteristic travel distance (CTD) of several thousand chemicals from the Canadian Domestic Substances List (DSL) and all current POPs using the multimedia model ELPOS. This shows that the CTD in water dominates the CTD in air only for chemicals that are characterized by a large half-life in water and a low air-water partition coefficient (about 38% of the nonionic organic substances selected from the DSL). In particular, there are substances that are not classified as persistent compounds in water but exhibit higher CTDs for transport in water than for transport in air. Finally, we evaluated whether the LRTP boundary derived from POP reference chemicals has to be revised if LRTP in water is included and found that this boundary can be applied to all organic chemicals regardless of their transport in air or water.

Environmental factors affecting the levels of legacy pesticides in the airshed of Delaware and Chesapeake Bays, USA

Organochlorine insecticides and their degradation products contribute to toxicity in Chesapeake Bay, USA, sediments and affect the reproductive health of avian species in the region; however, little is known of atmospheric sources or temporal trends in concentrations of these chemicals. Weekly air (n = 265) and daily rain samples (n = 494) were collected over 2000 to 2003 from three locations in the Delmarva Peninsula, USA. Pesticides were consistently present in the gas phase with infrequent detection in the particle phase. Hexachlorocyclohexanes (HCHs) and cis- and trans-chlordane were detected most frequently (95-100%), and cis- and trans-nonachlor, oxychlordane, heptachlor, heptachlor epoxide, dieldrin, and 1-chloro-4-[2,2-dichloro-1-(4-chlorophenyl)ethenyl]benzene (4,4'-DDE) were also detected frequently. The highest mean air concentrations were for dieldrin (60-84 pg/m(3)), gamma-HCH (37-83 pg/m(3)), and 4,4'-DDE (16-80 pg/m(3)). Multiple regression analyses of air concentrations with temperature and wind conditions using modified Clausius-Clapeyron equations explained only 30 to 60% of the variability in concentration for most chemicals. Comparison of the air concentrations and enthalpy of air-surface exchange values at the three sites indicate sources of chlordanes and alpha-HCH sources are primarily from long-range transport. However, examination of chlordane isomer ratios indicates some local and regional contributions, and gamma-HCH, 4,4'-DDE, dieldrin, heptachlor, heptachlor epoxide, and oxychlordane also have local or regional sources, possibly from contaminated soils. Median rain sample volumes of 1 to 3 L led to infrequent detections in rain; however, average measured concentrations were 2 to 10 times higher than in the Great Lakes. Dissipation half-lives in air were well below 10 years for all chemicals and below published values for the Great Lakes except dieldrin, which did not decline during the sample period.

Modeling the overall persistence and environmental mobility of sulfur-containing polychlorinated organic compounds

Experimental data on partition coefficients and environmental half-lives of sulfur analogs of polychlorinated organic compounds are scarce. Consequently, little is known about their overall persistence and long-range transport potential, which are the most vital measures in the environmental exposure assessment. We performed Multimedia Modeling of environmental fate and transport to complement this paucity of scientific data. The main aim of our study was to investigate whether the sulfur analogs of polychlorinated dibenzo-p-dioxins, -dibenzofurans, and -diphenylethers are as environmentally persistent and/or mobile as their oxygen counterparts and to propose the environmental exposure-related classification of the examined sulfur compounds. Our study included all possible congeners of the sulfur analogs generated in a combinatorial approach. We predicted (1) lacking data on partition coefficients (log K OW, log K OA and log K AW) for oxygen and sulfur analogs using Quantitative Structure-Property Relationship (QSPR) modeling and (2) their half-lives in air, water, and soil using US EPA tool 'The PBT Profiler, v. 1.203 2006'. Subsequently, we introduced these results into multimedia mass balance model 'The OECD POV and LRTP Screening Tool, v. 2.2'. Our study revealed that log K OW and log K OA are increasing by constant values of 0.60 and 1.07, respectively, and the values of log K AW are decreasing by 0.90, whenever one oxygen atom in the carbon skeleton is replaced by sulfur. The persistence ranking performed by the PBT Profiler showed that PCDDs, PCDFs, PCDEs, and their sulfur analogs belong to one half-life class. The Multimedia Modeling by the means of 'The OECD POV and LRTP Screening Tool, v. 2.2' suggested that the long-range transport potential depends on the presence/absence of oxygen/sulfur atoms in particular molecules, their substitution pattern and the parent carbon skeleton. Sulfur analogs are generally less mobile than their oxygen analogs, but have similar overall persistence and much higher bioaccumulation potential. Thus, according to the classification of chemicals proposed by Klasmeier et al. (Environ Sci Technol 40:53-60, 2006), some of them show POP-like POV and LRTP characteristics while the rest shows POP-like P OV characteristics. The sulfur analogs of PCDDs, PCDFs, or PCDEs bring environmental mobility comparable with the risk related to the oxygen ones; they belong to the pollutants of 'highest' or 'intermediate' priority. Further studies that would verify the necessity to include the studied sulfur molecules in the international lists of high-priority environmental pollutants are recommended.

Using Information on Uncertainty to Improve Environmental Fate Modeling: A Case Study on DDT

Present and future concentrations of DDT in the environment are calculated with the global multimedia model CliMoChem. Monte Carlo simulations are used to assess the importance of uncertainties in substance property data, emission rates, and environmental parameters for model results. Uncertainties in the model results, expressed as 95% confidence intervals of DDT concentrations in various environmental media, in different geographical locations, and at different points in time are typically between 1 and 2 orders of magnitude. An analysis of rank correlations between model inputs and predicted DDT concentrations indicates that emission estimates and degradation rate constants, in particular in the atmosphere, are the most influential model inputs. For DDT levels in the Arctic, temperature dependencies of substance properties are also influential parameters. A Bayesian Monte Carlo approach is used to update uncertain model inputs based on measurements of DDT in the field. The updating procedure suggests a lower value for half-life in air and a reduced range of uncertainty for Kow of DDT. As could be expected, the Bayesian updating yields model results that are closer to observations, and model uncertainties have decreased. Sensitivity analysis and Bayesian Monte Carlo approach in combination provide new insight into important processes that govern the global fate and persistence of DDT in the environment.

DosT and DevS are oxygen‐switched kinases in Mycobacterium tuberculosis

Exposure of Mycobacterium tuberculosis to hypoxia is known to alter the expression of many genes, including ones thought to be involved in latency, via the transcription factor DevR (also called DosR). Two sensory kinases, DosT and DevS (also called DosS), control the activity of DevR. We show that, like DevS, DosT contains a heme cofactor within an N-terminal GAF domain. For full-length DosT and DevS, we determined the ligand-binding parameters and the rates of ATP reaction with the liganded and unliganded states. In both proteins, the heme state was coupled to the kinase such that the unliganded, CO-bound, and NO-bound forms were active, but the O(2)-bound form was inactive. Oxygen-bound DosT was unusually inert to oxidation to the ferric state (half life in air >60 h). Though the kinase activity of DosT was unaffected by NO, this ligand bound 5000 times more avidly than O(2) to DosT (K(d) [NO] approximately 5 nM versus K(d) [O(2)] = 26 microM). These results demonstrate direct and specific O(2) sensing by proteins in M. tuberculosis and identify for the first time a signal ligand for a sensory kinase from this organism. They also explain why exposure of M. tuberculosis to NO donors under aerobic conditions can give results identical to hypoxia, i.e., NO saturates DosT, preventing O(2) binding and yielding an active kinase.

Atmospheric Deposition of Current-Use and Historic-Use Pesticides in Snow at National Parks in the Western United States

The United States (U.S.) National Park Service has initiated research on the atmospheric deposition and fate of semi-volatile organic compounds in its alpine, sub-Arctic, and Arctic ecosystems in the Western U.S. Results for the analysis of pesticides in seasonal snowpack samples collected in spring 2003 from seven national parks are presented herein. From a target analyte list of 47 pesticides and degradation products, the most frequently detected current-use pesticides were dacthal, chlorpyrifos, endosulfan, and gamma-hexachlorocyclohexane, whereas the mostfrequently detected historic-use pesticides were dieldrin, alpha-hexachlorocyclohexane, chlordane, and hexachlorobenzene. Correlation analysis with latitude, temperature, elevation, particulate matter, and two indicators of regional pesticide use reveal that regional current and historic agricultural practices are largely responsible for the distribution of pesticides in the national parks in this study. Pesticide deposition in the Alaskan parks is attributed to long-range transport because there are no significant regional pesticide sources. The percentage of total pesticide concentration due to regional transport (%RT) was calculated for the other parks. %RT was highest at parks with higher regional cropland intensity and for pesticides with lower vapor pressures and shorter half-lives in air.

Exposure chamber measurements of mass transfer and partitioning at the plant/air interface.

Dynamic measures of air and vegetation concentrations in an exposure chamber and a two-box mass balance model are used to quantify factors that control the rate and extent of chemical partitioning between vegetation and the atmosphere. A continuous stirred flow-through exposure chamber was used to investigate the gas-phase transfer of pollutants between air and plants. A probabilistic two-compartment mass balance model of plant/air exchange within the exposure chamber was developed and used with measured concentrations from the chamber to simultaneously evaluate partitioning (Kpa), overall mass transfer across the plant/air interface (Upa), and loss rates in the atmosphere (Ra) and aboveground vegetation (Rp). The approach is demonstrated using mature Capsicum annuum (bell pepper) plants exposed to phenanthrene (PH), anthracene (AN), fluoranthene (FL) and pyrene (PY). Measured values of log Kpa (V[air]/V[fresh plant]) were 5.7, 5.7, 6.0, and 6.2 for PH, AN, FL, and PY, respectively. Values of Upa (m d(-1)) under the conditions of this study ranged from 42 for PH to 119 for FL. After correcting for wall effects, the estimated reaction half-lives in air were 3, 9, and 25 h for AN, FL and PY. Reaction half-lives in the plant compartment were 17, 6, 17, and 5 d for PH, AN, FL, and PY, respectively. The combined use of exposure chamber measurements and models provides a robust tool for simultaneously measuring several different transfer factors that are important for modeling the uptake of pollutants into vegetation.

Relationship of approaches and a tiered methodology for screening chemicals in the context of long-range transport

In many national and international initiatives, where thousands of chemicals are screened, the ability of a chemical to be transported over long distances is an important criterion in determining whether environmental concern is warranted. Preliminary screening can be conducted using: (1) effective travel distance (ETD); (2) characteristic travel distance (CTD); and/or (3) the degradation half-life in air. The CTD is the distance traveled before the concentration of a chemical in air is reduced by a factor of 50%, for example. Differences in the distance traveled associated with the environmental release medium of a chemical are taken into account the ETD measure. The ETD can be defined as the distance traveled before the concentration in a stated medium (air, water, soil or sediment) is reduced to a specified level for a given mass release rate to air, to water and/or to soil. However, despite their merits, the use of multimedia screening measures like the ETD and CTD remains inhibited by both the limited availability of degradation data (particularly for soils and sediments) and release pattern information. Preliminary screening in terms of the atmospheric degradation half-life is commonly the only practical option. In this paper, straightforward guidelines based on partitioning coefficients (Henry's law constant and octanol–water partitioning coefficient) are proposed to reduce the degradation data requirements of multimedia measures like the ETD and CTD. The values used in the guidelines reflect a quantifiable trade-off between data acquisition requirements and uncertainty. The relationship of the potential screening options (using all degradation data versus using only data identified as required in the CTD and ETD approaches; screening in terms of the degradation half-life in air versus the CTD versus the ETD) is derived and the consequences of the differences are illustrated. A three-tiered screening methodology is then proposed. This tiered methodology will result in significant savings in time and money in national and international screening initiatives.

Screening Chemicals for Persistence in the Environment

A method is suggested for rapid screening of chemicals for persistence in the environment. Physical−chemical equilibrium partitioning information between air, water, and octanol are used as a first screen to identify the media for which degradation half-lives are required and those for which half-lives may be unnecessary. An overall persistence under equilibrium conditions is then estimated using half-lives in air, water, soil, and sediment using a steady-state mass balance model. A graphical technique to identify the key half-lives is demonstrated using 233 chemicals. For chemicals of more extreme partitioning properties, some half-lives may not be needed.

Apodinitrogenase: purification, association with a 20-kilodalton protein, and activation by the iron-molybdenum cofactor in the absence of dinitrogenase reductase

The Azotobacter vinelandii mutant strain UW45 contains a mutation in the nifB gene and produces an inactive dinitrogenase protein that can be activated by the addition of purified iron-molybdenum cofactor (FeMoco). This FeMoco-deficient dinitrogenase (Apo I) has now been purified 96-fold to greater than 95% purity and is FeMoco-activatable to 2200 nmol of C2H2 reduced/(min.mg of protein). The Apo I complex was found to contain two molecules of a 20-kDa protein, in addition to the alpha 2 beta 2 tetramer found for isolated holodinitrogenase (Holo I). The Apo I complex contained 15 +/- 2 mol of Fe per mole, but no Mo. While the presence of dinitrogenase reductase caused a 2-fold stimulation in the activation of the purified Apo I complex by FeMoco, this enhancement resulted from the stabilization of Apo I by dinitrogenase reductase to the denaturing effects of N-methylformamide. When the activation was performed in the absence of N-methylformamide, there was no enhancement by dinitrogenase reductase alone or by dinitrogenase reductase-Mg-ATP complex. The Apo I complex is more sensitive to O2 than Holo I, with a half-life in air of 6 min; however, the addition of dithiothreitol to Apo I during the exposure to air (or after exposure) resulted in a half-life very similar to that seen for Holo I. This suggests that sulfhydryl(s) is (are) important for the FeMoco-activation reaction.