Two-dimensional NMR studies of size fractionated Suwannee River fulvic and humic acid reference.

Two-dimensional phase sensitive 13C,1H correlation spectra were applied to the investigation of substructures in size fractions obtained by tangential flow multistage ultrafiltration (MST-UF) of humic substances (HS) Suwannee River Fulvic Acid Reference (HS SR FA) and Suwannee River Humic Acid Reference (HS SR HA), purchased from the International Humic Substances Society (IHSS). After size fractionation with MST-UF the HS samples give well resolved two-dimensional 13C,1H-correlated NMR spectra which offer a great potential for substructure elucidation and even quantification. It is shown that low molecular size lignin moieties undergo demethylation of the methoxy groups, accompanied by removal of the phenylpropane side chains and subsequent reaggregation of the aromatic rings. These findings provide insight into the processes of lignin degradation. Only the fraction >100 kDa contains macromolecules that have spin-spin relaxation times too short for investigations employing NMR multipulse sequences.

A new two-dimensional pulse sequence for T(2)* measurements of protons in (13)C isotopomers.

A new two-dimensional pulse sequence for T(2)* measurement of protons directly coupled to (13)C spins is proposed. The sequence measures the tranverse relaxation time of heteronuclear proton single-quantum coherence under conditions of free precession and is therefore well suited to evaluate relaxation losses of proton magnetization during preparation delays of heteronuclear pulse experiments in analytical NMR. The relevant part of the pulse sequence can be inserted as a "building block" into any direct or inverse detecting H,C correlation pulse sequence if proton spin-spin relaxation is to be investigated. In this contribution, the building block is inserted into a HETCOR as well as into a HMQC pulse sequence. Experimental results for the HETCOR-based sequence are given.

Quantification of carbohydrate structures in size fractionated aquatic humic substances by two-dimensional nuclear magnetic resonance.

Two-dimensional phase sensitive C,H correlation spectra were successfully applied to the quantification of carbohydrate substructures in aquatic humic substance (HS) fractions obtained by tangential flow multistage ultrafiltration (TFMSTUF) of a selected bog water HS (HO13, German Research Program DFG-ROSIG) as well as a river HS (Suwannee River Fulvic Acid Reference of the International Humic Substances Society, IHSS). It turns out that after size fractionation the HS samples give very well resolved C,H-correlation spectra which offer a great potential for substructure quantification. Details of the combined substructure quantification technique, novel in HS characterization, are presented. The results of the combined procedure point out that carbohydrate moieties predominantly occur in higher molecular mass fractions (> 10 kDa) of isolated HS.

Determination of exchangeable protons in natural organic matter using a home-made hydrogen/carbon analyser.

A home made hydrogen/carbon analyser was used to determine the portion of exchangeable protons in aquatic humic substances. For this purpose, equal sample amounts were dissolved in H2O and D2O, respectively, dried and combusted in a stream of oxygen. The amount of water resulting from combustion was measured by an infrared detector which recorded the OH bending vibration of H2O. The bands stemming from HOD or D2O were not registered by the detection unit. Thus, combustion of organic samples containing exchangeable protons dissolved in D2O resulted in a significantly smaller signal compared to the signal observed for the same sample dissolved in H2O. The relative intensity loss of the H2O signal observed after combustion was used to derive the portion of exchangeable protons in a standard reference material, a humic substance isolated by the International Humic Substances Society (Suwannee River fulvic acid). According to this method about 20% of the sample protons could be identified as exchangeable protons. With regard to titration data the portion of protons bound to non acidic hydroxy functions could be estimated. The validity of this procedure was proved by combustion experiments using commercially available deuterated substances as well as organic model compounds dissolved in D2O and H2O, respectively.

A novel high-temperature (360 degrees C)/high-pressure (30 MPa) flow system for online sample digestion applied to ICP spectrometry.

A flow injection sample digestion system has been developed comprising an indirectly electrically heated Pt/Ir capillary. Such a capillary allows reaction temperatures of up to 360 degrees C and pressures of up to 30 MPa (300 bar) and withstands concentrated acids. This temperature is 130 degrees C to 160 degrees C higher compared to the operating temperatures of microwave heated flow systems. A combination of an ultrasonic nebulizer and membrane desolvator serves as an interface between the flow digestion system and an ICP/AES spectrometer. The membrane desolvator removes interfering gaseous digestion products so effectively before the sample stream enters the plasma that the measured residual carbon concentration falls in the region of the detection limit of ICP/OES measurements. Sewage sludge samples were digested using nitric acid and the elemental traces online determined. The detection limits related to the original dry substances amount to the lower microg/g range.