Increasing the sensitivity of asymmetrical flow field-flow fractionation: Slot outlet technique.

A new enrichment approach is described to improve the sensitivity of the asymmetrical flow field-flow fractionation, with and without ICPMS online coupling for elemental detection. For the slot outlet technique, a part of the laminar carrier stream is removed through an additional pump. This allowed an enrichment of the colloidal particles in the separation channel up to a factor of 14. Additional improvements of the separation efficiency permitted us to separate colloids with differences in their molecular masses of only 2 kDa. Different polymer standards, proteins, and synthesized tracer colloids, as well as real samples (wastewater, liquid manure, serums) were used to assess the performance of the new technique.

Immunization with soot from a non-combustion process provokes formation of antibodies against polycyclic aromatic hydrocarbons.

The emission of soot during combustion processes used in transportation, manufacturing, and power generation is of increasing concern because of its serious adverse health effects. In particular, the ability to modulate the immune system has recently been established. In the present investigation, an artificial soot sample that was prepared by fragmentation of acetylene in a laser-induced plasma was used as an antigen for the immunization of a rabbit. A highly sensitive competition curve in an indirect competitive ELISA using a benzo[a]pyrene-BSA conjugate as a coating antigen could be constructed for benzo[a]pyrene with an IC50 of 2.94 mug/l (11.65 nmol/l). In contrast to the high affinity, the soot antiserum dilution (antibody titer) of 1:750 was rather low. The cross-reactivity was tested with 16 parent polycyclic aromatic hydrocarbons, 7 nitrated polycyclic aromatics, and 3 methylated, hydroxylated or butyric acid derivatives. The results obtained suggest that the vertebrate immune system can respond to an immunization with soot by the generation of high affinity IgG class antibodies against polycyclic aromatics. It is likely that antibodies are raised against the molecular structures which form the framework of the soot particles and not against adsorbed and extractable polyaromatic compounds. The experiments suggest that if soot is considered a T-independent antigen, the isotype switch, essentially from IgM to IgG, could have been caused by co-inoculation with a T-dependent antigen, i.e., mycobacteria contained in Freunds complete adjuvant. However, at the cellular level the mechanism remains to be uncovered.

Characterization of sewage plant hydrocolloids using asymmetrical flow field-flow fractionation and ICP-mass spectrometry.

Asymmetrical flow field-flow fractionation (AF4) was applied to characterize aquatic colloids from biological sewage plants and to infer information of colloidal loads, sources, and sinks within the plants, resp. the colloidal interaction with the aqueous phase and the sewage sludge. To characterize the colloids further, especially the distributions of colloid associated heavy metals, the AF4 system was coupled to an inductively coupled plasma mass spectrometer (ICP-MS). The size distribution is determined by AF4 with UV absorbance and fluorescence detection after a calibration by monodisperse polystyrene sulfonate standards (PSS). Samples from different sewage plants and from different depths and locations within a plant were compared. The fulvic/humic acid fraction with a particle diameter d(p) < 10 nm appeared to be comparable in all samples and decreases only slightly along the plants, whereas larger colloids with d(p) > 10 nm almost completely passed into the sewage sludge. The concentrations of the initial colloidal heavy metals decreased along the plants.