A new method for phosphate purification for oxygen isotope ratio analysis in freshwater and soil extracts using solid-phase extraction with zirconium-loaded resin.

RATIONALE: Phosphate (PO4 ) oxygen isotope (δ18 OPO4 ) analysis is increasingly applied to elucidate phosphorus cycling. Due to its usefulness, analytical methods continue to be developed and improved to increase processing efficiency and applicability to various sample types. A new pretreatment procedure to obtain clean Ag3 PO4 using solid-phase extraction (SPE) with zirconium-loaded resin (ZrME), which can selectively adsorb PO4 , is presented and evaluated here. METHODS: Our method comprises (1) PO4 concentration, (2) PO4 separation by SPE, (3) cation removal, (4) Cl- removal, and (5) formation of Ag3 PO4 . The method was tested by comparing the resulting δ18 OPO4 of KH2 PO4 reagent, soil extracts (NaHCO3 , NaOH, and HCl), freshwater, and seawater with data obtained using a conventional pretreatment method. RESULTS: PO4 recovery of our method ranged from 79.2% to 97.8% for KH2 PO4 , soil extracts, and freshwater. Although the recovery rate indicated incomplete desorption of PO4 from the ZrME columns, our method produced high-purity Ag3 PO4 and accurate δ18 OPO4 values (i.e., consistent with those obtained using conventional pretreatment methods). However, for seawater, the PO4 recovery was low (1.1%), probably due to the high concentrations of F- and SO4 2- which interfere with PO4 adsorption on the columns. Experiments indicate that the ZrME columns could be regenerated and used repeatedly at least three times. CONCLUSIONS: We demonstrated the utility of ZrME for purification of PO4 from freshwater and soil extracts for δ18 OPO4 analysis. Multiple samples could be processed in three days using this method, increasing sample throughput and potentially facilitating more widespread use of δ18 OPO4 analysis to deepen our understanding of phosphorus cycling in natural environments.

Sensitive Gas Chromatography Detection of Nanomolar Hydroxylamine in Environmental Water by Fe(III) Oxidation.

Nanomolar concentrations of NH2OH in natural water sources were determined using an Fe3+ oxidation method. A pH of 2.35 - 2.50 was used, which was adjusted by adding a chloroacetate buffer. Equal amounts (1.0 mL) of the chloroacetate solution and ferric chloride solution were added to the water sample (70 mL) to oxidize NH2OH to N2O. The resulting N2O in the sample water was then quantified by headspace analysis using a gas chromatograph with an electron-capture detector (ECD), where a limit of detection of 0.2 µgN L-1 (14 nmol L-1) was achieved. This method was successfully applied to samples of freshwater, brackish water, and seawater, and despite the various salinities no interfering substances were observed. Furthermore, NH2OH was successfully detected in samples collected from the Hii River and Lakes Shinji and Nakaumi (Shimane Prefecture, Japan). In addition, the proposed method was also applicable to samples rich in organic substance derived from phytoplankton.

Sensitive Method for the Oxidation-determination of Trace Hydroxylamine in Environmental Water Using Hypochlorite Followed by Gas Chromatography.

We developed a method for quantifying trace NH2OH in brackish- and sea-water samples. Previously reported methods applicable to fresh water cannot be applied to such samples. We determined that interference in seawater owing to the bromide ion can be removed by the addition of phenol. In our procedure, phenol and hypochlorite solutions were added to a sample solution to oxidize NH2OH to N2O. N2O in the sample was then quantified by headspace analysis. The method is not affected by the salt content or ammonia, nitrate, or nitrite at concentrations of 300 µgN L-1 or less. It has a limit of detection of 0.2 µgN L-1, and can quantify NH2OH in natural water samples with a wide range of salinity. It was applied to samples from Lake Nakaumi, a brackish lake located in the eastern part of Shimane Prefecture, Japan.

Determination of Trace Hydrazine in Environmental Water Samples by in situ Solid Phase Extraction.

A simple and rapid in situ method for the determination of hydrazine based on the concentration of aldazine compound formed by the reaction of hydrazine with p-dimethylaminobenzaldehyde was developed. This method was based on solid-phase extraction using a Sep-Pak C18 cartridge, followed by the quantification of hydrazine using a spectrophotometric method. To a sample solution of environmental water, p-dimethylaminobenzaldehyde solution was added to form aldazine by the reaction with hydrazine. The solution was passed through a Sep-Pak C18 cartridge for the adsorption of aldazine. In the laboratory, the aldazine adsorbed on the Sep-Pak C18 cartridge was eluted by passing a hydrochloric acid-ethanol (1:10) solution through the cartridge, and the color intensity of the solution was measured at 457 nm. The limit of detection for the new method was 0.2 mgN L-1 of hydrazine. The determination of hydrazine in solution was not influenced even by hydrogen sulfide and organic matter. This method was then applied to the brackish water of Lake Nakaumi in the eastern area of Shimane Prefecture, Japan. This method was used to determine hydrazine in freshwater, seawater and wastwater.

High spatial resolution analysis of the distribution of sulfate reduction and sulfide oxidation in hypoxic sediment in a eutrophic estuary.

Bottom hypoxia and consequential hydrogen sulfide (H2S) release from sediment in eutrophic estuaries is a major global environmental issue. We investigated dissolved oxygen, pH and H2S concentration profiles with microsensors and by sectioning sediment cores followed by colorimetric analysis. The results of these analyses were then compared with the physicochemical properties of the bottom water and sediment samples to determine their relationships with H2S production in sediment. High organic matter and fine particle composition of the sediment reduced the oxidation-reduction potential, stimulating H2S production. Use of a microsensor enabled measurement of H2S concentration profiles with submillimetre resolution, whereas the conventional sediment-sectioning method gave H2S measurements with a spatial resolution of 10 mm. Furthermore, microsensor measurements revealed H2S consumption occurring at the sediment surface in both the microbial mat and the sediment anoxic layer, which were not observed with sectioning. This H2S consumption prevented H2S release into the overlying water. However, the microsensor measurements had the potential to underestimate H2S concentrations. We propose that a combination of several techniques to measure microbial activity and determine its relationships with physicochemical properties of the sediment is essential to understanding the sulfur cycle under hypoxic conditions in eutrophic sediments.

Colorimetric Determination of Sulfide in Microsamples.

A method for determination of hydrogen sulfide in microsamples (200 µL) was developed by modifying the methylene blue method. Samples were collected using a micropipette and were combined with sulfide coloring reagent and 5 mL of 0.1 M HCl in test tubes. Absorbance of the solution was measured spectrophotometrically at 667 nm. This modified method did not require any special labware or technique, and can be used in a variety of research fields.