Microplastics in different water samples (seawater, freshwater, and wastewater): Methodology approach for characterization using micro-FTIR spectroscopy.

Microplastics of millimeter dimensions have been widely investigated in environmental compartments and today, studies are mainly focused on particles of smaller dimensions (< 500 µm). However, as there are no relevant standards or policies for the preparation and analysis of complex water samples containing such particles, the results may be questionable. Therefore, a methodological approach for 10 µm to 500 µm microplastic analysis was developed using µ-FTIR spectroscopy coupled with the siMPle analytical software. This was undertaken on different water samples (sea, fresh, and wastewater) taking into consideration rinsing water, digestion protocols, collection of microplastics, and sample characteristics. Ultrapure water was the optimal rinsing water and ethanol was also proposed with a mandatory previous filtration. Although water quality could give some guidelines for the selection of digestion protocols, it is not the only decisive factor. The methodology approach by µ-FTIR spectroscopy was finally assessed to be effective and reliable. This improved quantitative and qualitative analytical methodology for microplastic detection can then be used to assess the removal efficiency of conventional and membrane treatment processes in different water treatment plants.

Microplastics in different water samples (seawater, freshwater, and wastewater): Removal efficiency of membrane treatment processes.

The distribution and fate of microplastics in different water sources and their treatment plants (seawater, three municipal wastewaters, a pharmaceutical factory wastewater, and three drinking waters) in France were studied. Currently, research in this field is still under exploration since almost no relevant standards or policies have been introduced for the detection, the removal, or the discharge of microplastics. This study used an improved quantitative and qualitative analytical methodology for microplastic detection by µ-FTIR carried out with siMPle analytical software. By investigation, wastewater was determined to contain the most abundant microplastics in quantity (4,203-42,000 MP·L-1), then followed by surface water/groundwater (153-19,836 MP·L-1) and seawater (around 420 MP·L-1). Polyethylene was the dominant material in almost all water types followed by polypropylene, polystyrene, and polyethylene terephthalate. Almost all treatment technologies could remove microplastics whatever the feed water types and concentration of microplastics, though some treatment processes or transport pipes could cause additional contamination from microplastics. The four WWTPs, three DWTPs, and SWTP in France provided, respectively, 87.8-99.8%, 82.3-99.9%, 69.0-96.0% removal/retention of MPs in quantity, and provided 97.3-100%, 91.9-99.9%, 92.2-98.1% removal/retention of MPs in surface area. Moreover, ultrafiltration was confirmed to be an effective technology for microplastic retention and control of dimensions of microplastics in smaller ranges both in field-scale and lab-scale experiments. The 200 kDa ultrafiltration membrane could retain 70-100% and 80-100% of microplastics in quantity and in surface area, respectively.

Parameters optimization using experimental design for headspace solid phase micro-extraction analysis of short-chain chlorinated paraffins in waters under the European water framework directive.

The water framework directives (WFD 2000/60/EC and 2013/39/EU) force European countries to monitor the quality of their aquatic environment. Among the priority hazardous substances targeted by the WFD, short chain chlorinated paraffins C10-C13 (SCCPs), still represent an analytical challenge, because few laboratories are nowadays able to analyze them. Moreover, an annual average quality standards as low as 0.4µgL(-1) was set for SCCPs in surface water. Therefore, to test for compliance, the implementation of sensitive and reliable analysis method of SCCPs in water are required. The aim of this work was to address this issue by evaluating automated solid phase micro-extraction (SPME) combined on line with gas chromatography-electron capture negative ionization mass spectrometry (GC/ECNI-MS). Fiber polymer, extraction mode, ionic strength, extraction temperature and time were the most significant thermodynamic and kinetic parameters studied. To determine the suitable factors working ranges, the study of the extraction conditions was first carried out by using a classical one factor-at-a-time approach. Then a mixed level factorial 3×2(3) design was performed, in order to give rise to the most influent parameters and to estimate potential interactions effects between them. The most influent factors, i.e. extraction temperature and duration, were optimized by using a second experimental design, in order to maximize the chromatographic response. At the close of the study, a method involving headspace SPME (HS-SPME) coupled to GC/ECNI-MS is proposed. The optimum extraction conditions were sample temperature 90°C, extraction time 80min, with the PDMS 100µm fiber and desorption at 250°C during 2min. Linear response from 0.2ngmL(-1) to 10ngmL(-1) with r(2)=0.99 and limits of detection and quantification, respectively of 4pgmL(-1) and 120pgmL(-1) in MilliQ water, were achieved. The method proved to be applicable in different types of waters and show key advantages, such as simplicity, automation and sensitivity, required for the monitoring programs linked to the WFD.

Heavy metal concentrations in natural and human-impacted sediments of Segara Anakan Lagoon, Indonesia.

The concentrations of eight elements (Cr, Cu, Fe, Mn, Ni, Ti, V, and Zn) in surface sediments from Segara Anakan Nature Reserve (SARN), Indonesia, were determined using inductively coupled plasma-atomic emission spectroscopy following microwave-assisted acid digestion. In general, the heavy metal concentrations of the sediments were found to decrease in the sequence Fe > Ti > Mn > Zn > V > Cu > Cr > Ni. Sediment pollution assessment was carried out using a pollution status index contamination factor, pollution load index, geoaccumulation index, and enrichment factor as well as by comparing the measured values with two sediment quality guidelines, i.e., threshold effect level and probable effect level. The evaluation showed that in the refinery site stations, Cr, Ni, and Zn concentrations found in the SANR sediments may cause the adverse effect to occur over a wider range of organisms and can contribute to a more serious harmful effect.

Distributions and sources of persistent organic pollutants (aliphatic hydrocarbons, PAHs, PCBs and pesticides) in surface sediments of an industrialized urban river (Huveaune), France.

Surface sediments from the Huveaune River were analyzed for n-alkanes, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), organochlorine and organophosphorous pesticides (OCs and OPs) by using gas chromatography-mass spectrometry (GC-MS). Concentrations of total alkanes ranged from 184 to 26,780 µg·kg(-1) sediment dry weight (dw) with a mean concentration of 6,126 ± 8,006 µg·kg(-1)dw, concentrations of total PAHs ranged from 572 to 4,235 µg·kg(-1)dw with a mean concentration of 1966 ± 1,104 µg·kg(-1) dw, concentrations of total PCB ranged from 2.8 to 435 µg·kg(-1)dw with a mean concentration of 148 ± 164 µg·kg(-1)dw and concentrations of total pesticides ranged from 0.07 to 1.25 µg·kg(-1)dw with a mean concentration of 1.23 ± 1.29 µg·kg(-1)dw. The spatial distribution of POPs reveals that pollutant concentration is relatively higher at the mouth of the river. The molecular indices of specific n-alkanes (CPI, NAR and TAR) and molecular indices of PAHs (Ant/(Ant+Phe), Fl/(Fl+Pyr), BaA/(BaA+Chry), IPyr/(Ipyr+BghiP)) were calculated to evaluate the possible sources of hydrocarbons. These molecular indices suggest mainly pyrolytic inputs which are markedly biogenic. All contaminant levels were also compared with Sediments Quality Guidelines (SQG) showing that the contamination levels in all stations were most of the time lower than their respective SQG. While, for PCBs, five stations (H5, H6, H7, H8 and H9) were higher than their effect range median (ERM) values which may indicate high potential toxicity of the sediment with probable adverse effects to the living biota.

Quantitative and mid-infrared changes of humic substances from burned soils.

The humic substances are an abundant and important part of soil organic matter which plays many roles in ecosystems. On the other hand, forest fires are known to have a potential impact on the soil organic matter. Consequently, we chose to study the impact of forest fires on humic substances and the three relevant fractions, e.g. humic acids (HA), fulvic acids (FA) and non-humified fraction (NHF), NHF being the fulvic acids not adsorbed on XAD-8 resins. The studied site is a Mediterranean forest called "Maures Mountains", in France, where 30 sites were sampled in two layers: 0-5 and -5 to 15 cm. In order to validate the method, the 2 horizons from 5 sites randomly chosen were analyzed in triplicates. The extraction and fractionation procedures were achieved using alkali- and acid-solutions. The measurement of total organic carbon (TOC) using the TOC-meter and the use of Fourier transform infra-red (FTIR) spectroscopy gave us quantitative and qualitative results to evaluate the impact of forest fires and the role of their repetition. The results show that the fires led to significant decreases (Student test, P=0.05) of humic substances (HS), HA and of the fulvic fractions (FF=FA+NHF) in surface layers, corresponding to 40%, 34% and 35%, respectively. Moreover, the significant HA/FF ratio increases (Student test, P=0.05), as a result of the fire, indicate that NHF was probably transformed in FA-like compounds. About the qualitative impact, the results showed a significant decrease of alkyl and hydroxyl groups (OH), carboxylic acids and carbohydrates in HA after a wildfire, whereas the decrease was significant only for alkyl groups and carboxylic acids in FA. Lastly, the design of this work (control and burned sites, number of samples, time elapsed after fires, etc.) enables one to show the recovery of the Mediterranean forest ecosystem. FA quality and OH groups in HA are recovered between 3 and 16 years after the fire whereas alkyl groups, carboxylic acids and carbohydrates in HA as well as HS contents are not returned to the control state 16 years after the last fire.

Kinetic parameters for 17alpha-ethinylestradiol removal by nitrifying activated sludge developed in a membrane bioreactor.

The synthetic hormone 17alpha-ethinylestradiol (EE2) is primarily removed in wastewater treatment plants (WWTPs) by sorption, and nitrifying biomass has been shown to be responsible for EE2 biodegradation. Membrane bioreactor (MBR) technology was chosen to develop a community of autotrophic, nitrifying micro-organisms and determine kinetic parameters for EE2 biodegradation. Biological inhibition by azide was applied to differentiate sorption from biodegradation. Activated sludge (AS) was acclimated in the MBR to a substrate specific to autotrophic biomass and resulted in an increase in nitrifying activity. Acclimated AS was used to successfully biodegrade EE2 (11% increase in EE2 removal), and the overall removal of EE2 was determined to be 99% (sorption+biodegradation). AS used directly from a WWTP without acclimation removed EE2 only through sorption (88% removal of EE2). Therefore, higher nitrifying activity developed by acclimating AS allowed almost complete removal of EE2.

Fire impact on forest soils evaluated using near-infrared spectroscopy and multivariate calibration.

The assessment of physico-chemical properties in forest soils affected by fires was evaluated using near infrared reflectance (NIR) spectroscopy coupled with chemometric methods. In order to describe the soil properties, measurements were taken of the total organic carbon on solid phase, the total nitrogen content, the organic carbon and the specific absorbences at 254 and 280 nm of humic substances, organic carbon in humic and fulvic acids, concentrations of NH(4)(+), Ca(2+), Mg(2+), K(+) and phosphorus in addition to NIR spectra. Then, a fire recurrence index was defined and calculated according to the different fires extents affecting soils. This calculation includes the occurrence of fires as well as the time elapsed since the last fire. This study shows that NIR spectroscopy could be considered as a tool for soil monitoring, particularly for the quantitative prediction of the total organic carbon, total nitrogen content, organic carbon in humic substances, concentrations of phosphorus, Mg(2+), Ca(2+) and NH(4)(+) and humic substances UVSA(254). Further validation in this field is necessary however, to try and make successful predictions of K(+), organic carbon in humic and fulvic acids and the humic substances UVSA(280). Moreover, NIR coupled with PLS can also be useful to predict the fire recurrence index in order to determine the spatial variability. Also this method can be used to map more or less burned areas and possibly to apply adequate rehabilitation techniques, like soil litter reconstitution with organic enrichments (industrial composts) or reforestation. Finally, the proposed recurrence index can be considered representative of the state of the soils.

Monitoring of the evolution of an industrial compost and prediction of some compost properties by NIR spectroscopy.

Sewage treatment plants produce wastes resulting from the organic matter concentration in the form of sludge. A way of jointly treating and exploiting these increasing wastes jointly is the composting. Composting makes it possible to reduce volumes and the masses of wastes all while developing them in a product usable like organic soil enrichment. In this work, the composting process of an industrial sewage sludge composting plant was monitored to study the evolution of different physico-chemical parameters (temperature, moisture, pH, organic carbon, organic and inorganic nitrogen, organic carbon/organic nitrogen ratio, humic substances) and biochemical parameters (soluble fraction, hemicellulose, cellulose, lignin). Because these analyses are expensive and time consuming, we wanted to develop an alternative method to determine the maturity of compost related to compost properties with raw samples. Acceptable predictions were found for moisture, temperature, pH, organic carbon, organic carbon/organic nitrogen ratio, total-, organic- and ammoniacal nitrogen, fulvic- and humic acids and fulvic acids/humic acids ratio, but the error values were too high for the compost age to consider a quantification model. With regard to the biochemical parameters, this study is rather a preliminary test which shows the interest of the approach, but requires to be continued. Finally, the age of compost can be evaluated with Principal Component Analysis applied to NIR spectra.

An experimental design approach for optimizing polycyclic aromatic hydrocarbon analysis in contaminated soil by pyrolyser-gas chromatography-mass spectrometry.

Pyrolyser-gas chromatography-mass spectrometry was used to analyze polycyclic aromatic hydrocarbons in contaminated soil without preliminary extraction. Experimental research methodology was used to obtain optimal performance of the system. After determination of the main factors (desorption time, Curie point temperature, carrier gas flow), modeling was done using a Box-Behnken matrix. Study of the response surface led to factor values that optimize the experimental response and achieve better chromatographic results.

Effects of crude oil on phospholipid fatty acid compositions of marine hydrocarbon degraders: estimation of the bacterial membrane fluidity.

In this study, we investigated, in vitro, the effects of petroleum hydrocarbons on the phospholipid ester-linked fatty acid composition of Corynebacterium sp. Strain 8. The usual ratio of monounsaturated fatty acids E/Z (or trans/cis) was calculated. This ratio led to unexpected results because we found similar values for growths on either a hydrophobic substrate (crude oil) or a soluble carbon source (rich medium). The use of such an indicator seemed limited for monitoring an environmental stress, so we proposed an index based on the homeoviscous adaptation theory. A membrane viscosity index was defined and applied to Corynebacterium sp. Strain 8 (in vitro growth) and to a sedimentary community (in situ experiment). The results allowed us to estimate the membrane fluidity of both an isolated strain and a bacterial community in accordance with the medium hydrophobicity.

Comparison of n-eicosane and phenanthrene removal by pure and mixed cultures of two marine bacteria.

The biotransformation activities of two hydrocarbonoclastic marine bacteria, Corynebacterium sp. and Sphingomonas sp. 2MPII, on n-eicosane and phenanthrene were investigated. During a 56-day experiment, in pure and mixed cultures, Corynebacterium sp. and Sphingomonas sp. 2MPII removed about 70% of the initial n-eicosane and phenanthrene concentrations (1 and 0.4 g L(-1), respectively). In pure cultures, culturable cell abundances increased over time, from 0.8 to 8.6 x 10(-11) CFU L(-1) (Corynebacterium sp.) and from 2.1 to 16 x 10(-11) CFU L(-1) (Sphingomonas sp. 2MPII ) but remained barely constant in mixed cultures. We defined a biotransformation index based on the number of culturable cells rather than the culture protein content, with the biotransformation cell yield (BCY) expressed in grams hydrocarbon CFU(-1) per day to better characterize hydrocarbon removal in pure and mixed cultures. The BCY was markedly higher in mixed than in pure cultures, increasing by a factor of 2-10.7 and 2.3-4.7 for n-eicosane and phenanthrene removal, respectively.

Analysis of polycyclic aromatic hydrocarbons in contaminated soil by Curie point pyrolysis coupled to gas chromatography-mass spectrometry, an alternative to conventional methods.

Curie point pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) has been compared with classical extraction procedures (Soxhlet, sonication, KOH digestion, microwave-assisted) followed by GC-MS analysis for the determination of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil. In each case, the efficiency of the technique was examined for 16 PAHs included in the US Environmental Protection Agency Priority Pollutant List. The results indicate that the recovery of PAHs is dependent on the extraction technique. The highest recoveries of PAHs were obtained with Curie point pyrolysis and KOH digestion. Py-GC-MS appeared to be interesting alternative method for the determination of PAHs in contaminated soil. The results were validated by certified soil (CRM 104) analysis.

Influence of n-alkanes and petroleum on fatty acid composition of a hydrocarbonoclastic bacterium: Marinobacter hydrocarbonoclasticus strain 617.

This study concerns the effects of various long-chain n-alkanes, n-alkane mixtures and Arabian Light crude oil on the fatty acid (FA) composition of a sedimentary marine bacteria (Marinobacter hydrocarbonoclasticus strain 617), growing under aerobic conditions. The cultures with n-alkanes, as compared with soluble carbon sources, led to greater amounts of saturated and methyl branched FA (mainly belonging to a delta10 series). We observed the appearance or increase of saturated and unsaturated FA with the same carbon chain length (CCL) as the n-alkane carbon source (maximum for n-alkane CCL corresponding to the 'range' of the de novo synthesized fatty acids). We also observed a strong control of the oddness/evenness of the CCL of the FA by the oddness/evenness of the n-alkane. A n-alkane utilization index, (saturated + branched)/monounsaturated fatty acids (SAFA + BFA/MUFA) enabled discriminating between soluble carbon sources and hydrocarbons.

Subfractionation, characterization and photooxidation of crude oil resins.

Resins of five crude oils were obtained using SARA fractionation. The maltenic fraction of Blend Arabian Light, was further separated into six polar fractions. These fractions which are the constituents of the resins were analysed by FTIR spectroscopy. They appeared to be more oxidized, more aliphatic and less aromatic than asphaltenes. Photooxidation of resins showed that they are easily oxidizable and much more influenced by photooxidation than asphaltenes. The principal effect of photooxidation are: (i) increase of carbonyl group amounts and particularly formation of carboxylic groups, (ii) oxidation of sulfoxide in sulfone group, (iii) oxidation of alkyl chains and of aromatic rings (quinone structures).

New approach to study of spilled crude oils using high resolution GC-MS (SIM) and metastable reaction monitoring GC-MS-MS.

Polycyclic aromatic hydrocarbons (PAHs) and geochemical biomarkers are good environmental markers to study the origin and evolution of an oil spill. To have access to the greatest number of molecular ratios, no fractionation of oil into aliphatic and aromatic compounds is made. Three analytical MS approaches are tested to analyze markers in this total hydrocarbon fraction: classical quadrupole GC-MS, high resolution GC-MS (HR GC-MS) and metastable reaction monitoring GC-MS-MS (MRM GC-MS-MS). This analytical approach is used to follow the evolution of PAHs in petroleum polluted mangrove soils over 8 years by using molecular ratios between polycyclic aromatic hydrocarbons and tri- and tetracyclic terpanes.

Isolation and characterization of a marine bacterium capable of utilizing 2-methylphenanthrene.

A marine bacterium isolated from a coastal hydrocarbon-polluted sediment has been described and attributed on the basis of its phenotypic and genotypic characteristics to the genus Sphingomonas sp. This strain was capable of using an alkylated phenanthrene 2-methylphenanthrene, as sole source of carbon and energy. In experiments, 2-methylphenanthrene (0.2 g/l) was added as crystals to the culture medium. After 5 days of aerobic growth at 30 degrees C, 70% was degraded and the complete dissipation occurred after 20 days. Furthermore, the strain could degrade various kinds of polyaromatic compounds, but failed to grow on aliphatic hydrocarbons.

Biodegradation of the (aliphatic + aromatic) fraction of Oural crude oil. Biomarker identification using GC/MS SIM and GC/MS/MS.

A simplified fraction of the Oural crude oil (aliphatic and aromatic hydrocarbons) was incubated in the presence of an hydrocarbonoclastic bacterial community isolated from a marine sediment highly contaminated by petroleum residue. The biodegradation has been carried out under aerobic conditions for 5 weeks and followed by FTIR, UV synchronous luminescence and GC/FID. Disappearance of the n-alkanes (2nd week), an important attack of the isoprenoïd compounds (5th week) and preferential alteration of monomethylated polyaromatics were observed. Concerning the biomarkers, the bicyclic alkanes and pentacyclic terpanes have been comparatively elucidated using GC/MS data. The identification of C(26) to C(29) steranes has required a most selective method, namely GC/MS/MS. Many molecular ratios based on GC/MS abundances were calculated, which showed good stability. Consequently, they can be used to determine the origin of a petroleum even one altered by biodegradation.

New unsaturated long-chain fatty acids in the phospholipids from the Axinellida sponges Trikentrion loeve and Pseudaxinella cf. lunaecharta.

In order to identify new structures and especially those involved as biosynthetic intermediates, the fatty acid composition of whole phospholipids from two Senegalese marine sponges from the order Axinellida, Trikentrion loeve and Pseudaxinella cf. lunaecharta, has been investigated by analytical gas chromatography and gas chromatography/mass spectrometry. Several new fatty acids were identified as methyl esters and N-acyl pyrrolidides, namely 16-eicosenoic, 11-tetracosenoic, 5-pentacosenoic, 11-hexacosenoic, 11-octacosenoic, 23-triacontenoic, 17,21-hexacosadienoic, 19,23-octacosadienoic, 9,23-triacontadienoic, 5,9,21-hexacosatrienoic, and 5,9,25-triacontatrienoic. Trikentrion loeve and P. cf. lunaecharta contain fifteen (25.7% of the total acid mixture) and thirteen (30.4%) delta 5,9 fatty acids, respectively. Based on gas chromatography/Fourier transform infrared experiments, the double bonds were assigned the (Z) configuration. Biosynthesis of dienoic and trienoic demospongic acids possessing an n-5 or an n-7 terminal double bond is discussed.

Hydrocarbons and fatty acids in surficial sediments (French Mediterranean coastal zones).

A detailed study has been made of hydrocarbons and fatty acids in nine surficial Mediterranean sediments. The hydrocarbon concentrations are generally low. The nature and the distribution of the hydrocarbons found in these sediments indicate that they originate from several different sources (terrestrial, biogenic, pyrolytic) and petroleum inputs are usually not the main source of hydrocarbons. Fatty acid concentrations vary in the range 2-52 mg.kg-1 dry sediment. Fatty acid distributions, between C14 and C20 show the biogenic autochthonous inputs. Branched and cyclopropanic fatty acids reflect a bacterial origin. These distributions are markedly different in oxic and anoxic sediments.