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.

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.

Survival and differentiation defects contribute to neutropenia in glucose-6-phosphatase-ß (G6PC3) deficiency in a model of mouse neutrophil granulocyte differentiation.

Differentiation of neutrophil granulocytes (neutrophils) occurs through several steps in the bone marrow and requires a coordinate regulation of factors determining survival and lineage-specific development. A number of genes are known whose deficiency disrupts neutrophil generation in humans and in mice. One of the proteins encoded by these genes, glucose-6-phosphatase-ß (G6PC3), is involved in glucose metabolism. G6PC3 deficiency causes neutropenia in humans and in mice, linked to enhanced apoptosis and ER stress. We used a model of conditional Hoxb8 expression to test molecular and functional differentiation as well as survival defects in neutrophils from G6PC3(-/-) mice. Progenitor lines were established and differentiated into neutrophils when Hoxb8 was turned off. G6PC3(-/-) progenitor cells underwent substantial apoptosis when differentiation was started. Transgenic expression of Bcl-XL rescued survival; however, Bcl-XL-protected differentiated cells showed reduced proliferation, immaturity and functional deficiency such as altered MAP kinase signaling and reduced cytokine secretion. Impaired glucose utilization was found and was associated with ER stress and apoptosis, associated with the upregulation of Bim and Bax; downregulation of Bim protected against apoptosis during differentiation. ER-stress further caused a profound loss of expression and secretion of the main neutrophil product neutrophil elastase during differentiation. Transplantation of wild-type Hoxb8-progenitor cells into irradiated mice allowed differentiation into neutrophils in the bone marrow in vivo. Transplantation of G6PC3(-/-) cells yielded few mature neutrophils in bone marrow and peripheral blood. Transgenic Bcl-XL permitted differentiation of G6PC3(-/-) cells in vivo. However, functional deficiencies and differentiation abnormalities remained. Differentiation of macrophages from Hoxb8-dependent progenitors was only slightly disturbed. A combination of defects in differentiation and survival thus underlies neutropenia in G6PC3(-/-) deficiency, both originating from a reduced ability to utilize glucose. Hoxb8-dependent cells are a model to study differentiation and survival of the neutrophil lineage.

Picogram determination of "earthly-musty" odorous compounds in water using modified closed loop stripping analysis and large volume injection GC/MS.

"Earthy-musty" off-flavor problems in water samples are due to organic compounds present at the sub-part-per-trillion level. Numerous analytical methods such as purge and trap, liquid/liquid extraction, and closed-loop stripping analysis (CLSA) followed by GC/MS analysis have been used to determine these compounds. However, these methods offer poor sensitivity (detection limits of approximately 1 to 10 ng/L) when compared to the 20-30 pg/L of sensorial sensitivity. The purpose of this study was to develop a new method involving a modified CLSA preconcentration technique together with large volume injection GC/MS in order to attain analytical sensitivity equal to or better than olfactory sensitivity. For eight target compounds that cause taste and odor problems in water at trace levels, the method developed was linear in the 0.05-10 ng/L range and provided recoveries greater than 70% together with satisfactory repeatability. Detection limits as low as 15-30 pg/L were achieved, representing a 50-fold improvement in sensitivity as compared to current methods. The accuracy and sensitivity of the method were demonstrated in different aqueous matrixes, including raw surface water. The method was successfully applied to earthy-musty water samples that had remained unsolved by conventional techniques, thus proving its effectiveness.