Determination of endocrine-disrupting phenolic compounds and estrogens in surface and drinking water by HRGC-(NCI)-MS in the picogram per liter range.

A method for the analysis of phenolic estrogenic active compounds in surface and drinking water in the picogram per liter range is described. Besides the widely used monomer bisphenol A, 4-tert-octylphenol [4-(1,1,3,3-tetramethylbutyl)phenol] and the technical isomer mixture of 4-nonylphenol; phenolic steroid hormones such as the endogenous estrogens estrone, 17alpha-estradiol, and 17beta-estradiol; and the exogenous estrogen 17alpha-ethinylestradiol were determined in water at the 20-200 pg/L level. Water samples from 1 to 5 L were extracted by solid-phase extraction (SPE) on a cartridge system containing LiChrolut EN as sorbent. The phenols and steroids were converted into their pentafluorobenzoylate esters in an extractive derivatization reaction. The derivatives were then determined by high-resolution gas chromatography with negative chemical ionization mass spectrometric detection (HRGC-(NCI)-MS) in the selected ion mode (SIM). All results were also confirmed by HRGC with electron capture detection (ECD). This highly sensitive and specific method gives a limit of detection (LOD) of 20 pg/L for bisphenol A and 4-tert-octylphenol in drinking water samples and 50 pg/L in STW effluent, respectively. The LODs for technical 4-nonylphenol, 17alpha-ethinylestradiol, and other estrogens are in the range of 50 pg/L in drinking water to 200 pg/L in STW effluent, respectively. In all river water samples in southern Germany, bisphenol A was found in concentrations ranging from 500 pg/L up to 16 ng/L, 4-nonylphenol was from 6 up to 135 ng/L, and the steroids were from 200 pg/L up to 5 ng/L. In drinking water, bisphenol A was found in concentrations ranging from 300 pg/L to 2 ng/L, 4-nonylphenol was from 2 to 15 ng/L, 4-tert-octylphenol was from 150 pg/L to 5 ng/L, and the steroids were from 100 pg/L to 2 ng/L. Mean recoveries over the whole analytical protocol, measured in bidistilled water, generally exceeded 70%. These results indicate that environmental endocrine-disrupting estrogens are not completely removed in the process of sewage treatment but are carried over into the general aquatic environment. After ground passage, they can eventually be found in drinking water.

Halogenated methyl-phenyl ethers (anisoles) in the environment: determination of vapor pressures, aqueous solubilities, Henry's law constants, and gas/water- (Kgw), n-octanol/water- (Kow) and gas/n-octanol (Kgo) partition coefficients.

Halogenated methyl-phenyl ethers (methoxybenzenes, anisoles) are ubiquitous organics in the environment although they are not produced in industrial quantities. Modelling the fate of organic pollutants such as halogenated anisoles requires a knowledge of the fundamental physico-chemical properties of these compounds. The isomer-specific separation and detection of 60 of the 134 possible congeners allowing an environmental fingerprinting are reported in this study. The vapor pressure p0(L) of more than 60 and further physico-chemical properties of 26 available congeners are given. Vapor pressures p0(L), water solubilities S(L)W, and n-octanol/water partition coefficients Kow were determined by capillary HR-GC (High Resolution Gas Chromatography) on a non-polar phase and by RP-HPLC (Reversed Phase High Performance Liquid Chromatography) on a C18 phase with chlorobenzenes as reference standards. From these experimental data the Henry's law constants H, and the gas/water Kgw and gas/n-octanol Kgo partition coefficients were calculated. We found that vapor pressures, water solubilities, and n-octanol/water partition coefficients of the halogenated anisoles are close to those of the chlorobenzenes. A similar environmental fate of both groups can, therefore, be predicted.

Characterisation of five technical mixtures of brominated flame retardants.

The main components of five technical mixtures of brominated flame retardants were identified by mass spectroscopy, H-NMR, IR spectroscopy, elementary analysis, and HRGC-MS, respectively. The mixtures have been identified as: phosphoric acid bromopropylates: Bromcal P 67-6 HP; C18-alkylated bromophenols: Bromcal P 40-3; dibromopropyl-2,4,6-tribromophenyl ether: Bromcal 73-5 PE; brominated diphenyl ethers (PBrDE): Bromcal 70-5 DE; decabromodiphenyl ether: Bromcal 82-0. Gas chromatographic retention data of the various constituents were measured on standard GC columns. The data prove the interference of some of the brominated compounds with PCBs and other halogenated pesticides in HRGC using the electron capture detector.

The deep-sea as a final global sink of semivolatile persistent organic pollutants? Part I: PCBs in surface and deep-sea dwelling fish of the north and south Atlantic and the Monterey Bay Canyon (California).

The understanding of the global environmental multiphase distribution of persistent organic pollutants (POPs) as a result of the physico-chemical properties of the respective compounds is well established. We have analysed the results of a vertical transport of POPs from upper water layers (0-200 m) to the deepwater region (> 800 m) in terms of the contamination of the biophase in both water layers. The contents of persistent organochlorine compounds like polychlorinated biphenyls (PCBs) in fish living in the upper water layers of the North Atlantic and the South Atlantic, and at the continental shelf of California (Marine Sanctuary Monterey Bay and its deep-sea Canyon) are compared to the levels in deep-sea or bottom dwelling fish within the same geographic area. The deep-sea biota show significantly higher burdens as compared to surface-living species of the same region. There are also indications for recycling processes of POPs--in this case the PCBs--in the biophase of the abyss as well. It can be concluded that the bio- and geo phase of the deep-sea may act similarly as the upper horizons of forest and grasslands on the continents as an ultimate global sink for POPs in the marine environment.

The deep-sea as a final global sink of semivolatile persistent organic pollutants? Part II: Organochlorine pesticides in surface and deep-sea dwelling fish of the north and south Atlantic and the Monterey Bay Canyon (California).

The understanding of the global environmental multiphase distribution of persistent organic pollutants (POPs) as a result of the physico-chemical properties of the respective compounds is well established. We have analysed the results of a vertical transport of POPs from surface water to deepwater in terms of the contamination of the biota living in the respective environmental compartments. Samples were taken from the North and the South Atlantic and from the uprising water region of the continental shelf of California (Marine Sanctuary Monterey Bay and its Canyon). The contents of persistent organochlorine pesticides (DDTs, chlordanes, toxaphenes, HCHs, and HCB) in surface-living fish are compared to those in deepwater fish of the same geographic area. The deepwater biota show significantly higher burdens as compared to surface-living species of the same region. There are also indications for recycling processes of POPs of the class of organochlorine pesticides in the biophase of the abyss as well. It can be concluded that the bio- and geophase of the deep-sea may act as an ultimate global sink for persistent semivolatile contaminants in the marine environment like the soil on the continents.

A new sample preparation technique for organochlorines in cod liver oil combining SPE and NP-HPLC with HRGC-ECD.

The analysis of semivolatile organochlorines (polychlorinated biphenyls and chlorinated pesticides) in less polluted biomaterials requires specific strategies in controlling the blank in sample preparation. The procedure described here allows to decrease significantly the level of contamination during the clean-up step of fish oil. Solid-phase-extraction (SPE) on LiChrolut EN and normal phase HPLC in the normal- and the backflush-mode were used to reduce the amount of solvents needed and the analysis time compared to established clean-up procedures. With a certified reference material (BCR-CRM 349; Cod Liver Oil) the precision and effectiveness of the new method were validated. Recovery rates of the Internal Standards (PCB 103 and TCN) lay between 75% and 90% at the microg/kg lipid level. The quantitative analyses were carried out by high resolution gas chromatography with electron capture detector (HRGC-ECD).

New stationary phase based on beta-cyclodextrin for normal-phase HPLC group-separation of organic nitrates.

The synthesis of a beta-cyclodextrin-silica normal-phase (NP)-liquid chromatography (LC) stationary phase is reported. Silica gel was modified with (3-bromopropyl)trimethoxysilane to a 3-bromopropyl-silica that was reacted with beta-cyclodextrin, resulting in a beta-cyclodextrin-silica. To prove its usefulness in group-separation of organic nitrates among others, a mixture of three groups of organic nitrates was separated. The results are compared with those obtained on a nitrated polyol-silica that has recently been reported. The alkyl dinitrates exhibt higher retention relatively to alkyl mononitrates on the new phase. This allows to cut the LC fractions in a way that the alkyl mononitrates and phenylalkyl nitrates appear in one fraction and the dinitrates in a second one without any overlap of the two fractions.

Analysis of halogenated C1/C2-trace compounds in marine atmosphere.

Patterns of volatile ECD-active substances in marine air at the ultra trace level were investigated. The protocol used carbon based adsorbents, thermodesorption and thick film HRGC-ECD together with a second ECD in line with oxygen doped make-up gas. The latter ECD mode leads to an increased sensitivity for chloromethane. dichloromethane, and other important chlorinated substances with an otherwise low response at an ECD driven with oxygen-free nitrogen or argon/methane. The experimental set-up is easily to employ. Air samples were taken on two cruises of the German research vessel R/V "Polarstem" in the Atlantic Ocean to determine the background pattern of all volatile halocarbons from CF3Cl to CHI3 in marine air.

Determination of endogenous and exogenous estrogens in effluents from sewage treatment plants at the ng/L-level.

An analytical method for the determination of the major endogenous and exogenous estrogenic steriods in effluent water samples of sewage treatment plants (STPs) with a LOQ down to 1 ng/L and below has been developed. The exogenous estrogen 17alpha-ethynylestradiol, frequently used as estrogenic component in oral contraceptives, and the endogenous estrogen 17beta-estradiol show the highest estrogenic potential, therefore they were part of our target compounds. In addition, the content of the synthetic gestagen levonorgestrel, also often administered in oral contraceptives, was determined. A solid-phase extraction system for high volume sampling of water up to 25 L was implemented. Two types of adsorbent, Amberlite XAD 2 and a mixture of LiChrolut EN/Bondesil C-18, respectively, were tested for their extraction efficiency of these polar analytes. Recovery rates with LiChrolut EN/Bondesil C-18 ranged up to 94%, whereas sampling on XAD 2 led only to poor recoveries below 40%. After a liquid chromatographic clean-up step on silicagel the steroids were converted into their trimethylsilyl-ethers by the reaction with MSTFA/TMSI (N-methyl-N-trimethylsilyl-2,2,2-trifluoroacetamide, trimethylsilyliodide) and were then determined by HRGC/MS in the selected ion mode. A limit of quantification over the whole procedure of at least 1 ng/L was reached for all analytes. In several effluent samples the input of estrogens by the STP of the cities Ulm and New Ulm into the river Danube was characterised. The concentrations commonly found ranged from 1 ng/L up to 13 ng/L, depending on the respective steroid.

Multifunctional organic nitrates as constituents in European and US urban photo-smog.

Air samples of the atmospheric ground layer in the cities of Ulm in Germany, Las Vegas, Nevada, and Salt Lake City, Utah, in the US were analyzed for organic nitrates. The air samples were taken around noon in summer at sunny weather conditions. 43 (C6-C13) alkyl mononitrates, 24 (C3-C6) alkyl dinitrates, and 19 (C2-C6) hydroxy alkyl nitrates have been identified. The analytical protocol included high-volume-sampling, NP-HPLC group separation, high resolution capillary gas chromatography, and detection by the highly selective mass spectrometer detector (SIM mode, m/z = 46). The levels of the sum of 15 (C6-C10) alkyl mononitrates ranged from 2.9 to 11.0 parts per trillion (ppt(v)). The levels of the sum of 21 (C3-C6) alkyl dinitrates ranged from 2.3 to 10.5 ppt(v), and the levels of the sum of 7 (C2-C4) hydroxy alkyl nitrates ranged from 7.3 to 28 ppt(v), respectively, in the urban air samples. These results emphazise the contribution of the alkyl dinitrates and hydroxy alkyl nitrates besides the alkyl mononitrates to the budget of NO(Y) compounds. No major differences in levels and pattern of the organic nitrates are present in air of the German and the US cities.

Alkyl nitrates as achiral and chiral solute probes in gas chromatography. Novel properties of a beta-cyclodextrin derivative and characterization of its enantioselective forces.

Achiral and chiral interactions of alkyl nitrates (R-O-NO2) with heptakis-(3-O-acetyl-2,6-di-O-pentyl)-beta-cyclodextrin (LIPODEX-D) in the gas phase were investigated chromatographically. Two major outcomes can be summarized. First, LIPODEX-D shows very fast temperature-dependent variations of the selectivity up to changes in the order of elution for C1-C5 alkyl nitrates. These changes in selectivity reveal that LIPODEX-D possesses different shape selectivities for small alkyl nitrates at different temperatures (40-80 degrees C), i.e. with increasing isothermal separation temperature extended (chain-like) alkyl nitrates have increased retention relative to bulky alkyl nitrates. The observations are highly reproducible and might indicate conformational changes of the cyclodextrin, however, an ultimate proof would require further spectroscopic investigation. Secondly, the chiral separations of systematically varied sets of C4-C11 alkyl nitrates allowed the thermodynamic characterization of enantiodiscriminating interactions. Quantitative evidence is provided showing that the presence of an ethyl group at the asymmetric carbon atom of an n-alkyl nitrate gives a strong enthalpic contribution to the resulting enantioselectivity. The Gibbs free energy differences -deltaR,S(deltaG) decrease systematically three to six times if the ethyl group is either shortened or enlarged by only a -CH2-increment. The results are based on two separate thermodynamic approaches, i.e. the determination of thermodynamic quantities (-deltaR,S(deltaG), -deltaR,S(deltaH), -deltaR,S(deltaS), Tiso) and a theoretical concept of enthalpy-entropy compensation. The data from our laboratory experiments also indicate that van der Waals interactions are responsible for chiral discrimination.

C3-C14-alkyl nitrates in remote South Atlantic air

Alkyl nitrates with chain lengths up to fourteen carbons have been detected in the remote troposphere of the South Atlantic Ocean. The concentrations of numerous iso- and n-alkyl nitrates in the range of C3-C12 have been measured. Tenax-GC analysis was applied to determine short chain alkyl nitrates (< C6), while the novel detection of long chain iso- and n-alkyl nitrates was achieved with a high volume sampling technique using silica gel. C3-C12-alkyl nitrates show the lowest concentrations in the trade wind region (3.5 pptv). Higher concentrations in the west wind belt (11.1 pptv) reveal the influence of the South American continent as the source for the alkyl nitrates after long range transport. The concentrations of alkyl nitrates increase in proximity to South America. Measurements of long chain alkyl nitrates in rural air near the city of Ulm (Germany) are given for comparison. In addition, pattern analysis of long chain alkyl nitrates reveal this complex mixture of new compounds to be useful as trace indicators to distinguish continental and marine air masses. Despite the lower photochemical formation yields of primary n-alkyl nitrates compared to secondary n-alkyl nitrates, the primary n-alkyl nitrates are relatively increased in marine air. The reason for this finding cannot be explained so far, but the correlation of secondary/primary n-alkyl nitrates shows a significant differentiation of air samples with continental or marine character.

Vapour pressures, aqueous solubilities, Henry's law constants, partition coefficients between gas/water (Kgw), n-octanol/water (Kow) and gas/n-octanol (Kgo) of 106 polychlorinated diphenyl ethers (PCDE)

Modelling the environmental fate of persistent organic pollutants like polychlorinated diphenyl ethers (PCDE) requires the knowledge of a number of fundamental physico-chemical properties of these compounds. We report here the physico-chemical properties of 106 PCDEs, which are over 50% of all possible congeners. Vapour pressures P(OL), water solubilities S(H2O), and n-octanol/water partition coefficients K(OW) were determined with chromatographic methods. With these experimental data the Henry's law constants H, gas/water K(GW) and gas/n-octanol K(GO) partition coefficients were calculated. Vapour pressures and water solubilities and n-octanol/water partition coefficients of the PCDEs are close to those of similar groups of organochlorine compounds like polychlorinated biphenyls (PCBs) and dibenzofurans (PCDFs). A similar environmental fate can be predicted and was partially already been observed.

Determination of C1-C5 alkyl nitrates in rain, snow, white frost, lake, and tap water by a combined codistillation head-space gas chromatography technique. Determination of Henry's law constants by head-space GC.

Alkyl nitrates with a chain length up to five carbon atoms have been determined in snow, white frost, and surface water. The samples were taken in the vicinity of Ulm, Germany, a region in central Europe. The determination of C1-C5-alkyl nitrates in water samples was achieved with a new water codistillation enrichment technique directly coupled with on-column head-space gas chromatography. The concentrations of the short chain alkyl nitrates in the different forms of wet deposition range from 89 ng L-1 for 1-propyl nitrate down to 35 ng L-1 for 1-pentyl nitrate. C1-C5-alkyl nitrates in wet depositions were also directly determined by static head-space gas chromatography. Gas-water partition coefficients KGW (Henry's law constant H) were determined by head-space gas chromatography and secondly by calculating the Henry's law constant by the ratio of vapor pressure to water solubility. The gas-water partition constants (dimensionless) or Henry's law constants range from KGW = 0.038 (H = 93 Pa m3 mol-1) for 1-propyl nitrate up to KGW = 0.122 (H = 302 Pa m3 mol-1) for 2-pentyl nitrate.

Marine mammals as global pollution indicators for organochlorines.

Blubber tissues of the following marine mammals differing in their geographic distribution (North Atlantic, North Pacific, Bering Sea/Arctic Ocean), trophic level, and feeding habits were analyzed for their organochlorine contents: two seal species (harbor seals--Phoca vitulina, northern fur seals--Callorhinus ursinus), three toothed whale species (belukha whales--Delphinapterus leucas, one common dolphin--Delphinus delphis, one pilot whale--Globicephala melaena), and one baleen whale species (a bowhead whale--Balaena mysticetus). As xenobiotics were quantified the seven indicator congeners of the polychlorinated biphenyls (PCB 28, 52, 101, 118, 138, 153, and 180), three isomers of the hexachlorocyclohexanes (alpha-, beta- and gamma-HCH) as well as six components of the DDT-group (4,4'-DDT, 4,4'-DDD, 4,4'-DDE, 2,4'-DDT, 2,4'-DDD, and 2,4'-DDE). When comparing the xenobiotic levels of these marine mammals, it showed that the animals from the western North Atlantic were contaminated about 15 times more with organochlorines than the animals from the eastern North Pacific and the Bering Sea/Arctic Ocean. The total organochlorine burden, the 4,4'-DDE-percentage as well as the metabolic PCB patterns correlate with the trophic levels of the marine mammals studied. The quantitative analyses were done by high-resolution capillary gas chromatography with electron capture detection (HRGC/ECD) whereas the analyses of the metabolic PCB patterns were done by high-resolution capillary gas chromatography and mass selective detection (HRGC/MSD).

Interlaboratory comparison study for PCB congeners and chlorinated pesticides in beluga whale blubber.

Three laboratories participated in an interlaboratory comparison exercise for the determination of polychlorinated biphenyl (PCB) congeners and chlorinated pesticides in Standard Reference Material (SRM) 1588 (Organics in Cod Liver Oil) a whale blubber control material, and six beluga whale (Delphinapterus leucas) blubber samples. The results are generally in good agreement. The minor disagreement of some of the results may be attributed to differences in detector calibrations, differences in chromatographic separation selectivity for specific compounds, or both. The SRM and control material help to validate the analytical procedures and to verify that these procedures remain in control.

Cyclodextrin- and sugar-derivatives as stationary phases for the gas chromatography of semivolatiles.

Cyclodextrins as cyclic hexa-(alpha) to octa-(gamma) saccharides are widely and successfully used after esterification or etherification as stationary phases in gas chromatography for the separation of chiral compounds. Their basic potential as thermostable stationary phases as such or after derivatisation offers a unique spectrum of interactions ranging from hydrogen bonding to shape selectivity. Similar properties as thermostable stationary phases are shown by long-chain saccharide ethers (chain-length C8-C20), which are normally used as non-ionic detergents.

Determination of biogenic halogenated methyl-phenyl ethers (halogenated anisoles) in the picogram m(-3) range in air.

Halogenated anisoles (methyl-phenyl ethers) appear to be ubiquitous organic trace compounds in the environment. An analytical method is presented for analyzing the altogether 134 congeners of chloro-, bromo- and mixed bromochloro-anisoles on an isomer-specific basis in air. High volume sampling (20 m(3) per hour) is carried out by adsorption, using a mixture of silica gel 60 and ENVI-Carb. The preseparation of the anisoles is achieved by NP-LC on Florisil (a magnesium silicate). The interference by volatile n-alkyl nitrates in the electron-capture detection can be avoided by a LC-preseparation on a carboneous phase. The isomer-specific separation and detection is performed by HRGC-ECD and HRGC-MS-SIM. A structure-specific systematic numbering of the 134 congeners of the chloro-, bromo- and bromochloro-anisoles is suggested. The retention data are given of 16 chloroanisoles, 10 bromoanisoles and 27 bromo-chloroanisoles on three stationary phases with different polarity.