Geology topics

Charles J. Patton

Publications and source records attributed to Charles J. Patton.

7 recordsLinked to original sources

USGS2013

Analytical properties of some commercially available nitrate reductase enzymes evaluated as replacements for cadmium in automated, semiautomated, and manual colorimetric methods for determination of nitrate plus nitrite in water

A multiyear research effort at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) evaluated several commercially available nitrate reductase (NaR) enzymes as replacements for toxic cadmium in longstanding automated colorimetric air-segmented continuous-flow analyzer (CFA) methods for determining nitrate plus nitrite (NO x ) in water. This research culminated in USGS approved standard- and low-level enzymatic reduction, colorimetric automated discrete analyzer NO x methods that have been in routine operation at the NWQL since October 2011. The enzyme used in these methods (AtNaR2) is a product of recombinant expression of NaR from Arabidopsis thaliana (L.) Heynh. (mouseear cress) in the yeast Pichia pastoris. Because the scope of the validation report for these new automated discrete analyzer methods, published as U.S. Geological Survey Techniques and Methods 5–B8, was limited to performance benchmarks and operational details, extensive foundational research with different enzymes—primarily YNaR1, a product of recombinant expression of NaR from Pichia angusta in the yeast Pichia pastoris—remained unpublished until now. This report documents research and development at the NWQL that was foundational to development and validation of the discrete analyzer methods. It includes: (1) details of instrumentation used to acquire kinetics data for several NaR enzymes in the presence and absence of known or suspected inhibitors in relation to reaction temperature and reaction pH; and (2) validation results—method detection limits, precision and bias estimates, spike recoveries, and interference studies—for standard- and low-level automated colorimetric CFA-YNaR1 reduction NO x methods in relation to corresponding USGS approved CFA cadmium-reduction (CdR) NO x methods. The cornerstone of this validation is paired sample statistical and graphical analysis of NOx concentrations from more than 3,800 geographically and seasonally diverse surface-water and groundwater samples that were analyzed in parallel by CFA-CdR and CFA enzyme-reduction methods. Finally, (3) demonstration of a semiautomated batch procedure in which 2-milliliter analyzer cups or disposable spectrophotometer cuvettes serve as reaction vessels for enzymatic reduction of nitrate to nitrite prior to analytical determinations. After the reduction step, analyzer cups are loaded onto CFA, flow injection, or discrete analyzers for simple, rapid, automatic nitrite determinations. In the case of manual determinations, analysts dispense colorimetric reagents into cuvettes containing post-reduction samples, allow time for color to develop, insert cuvettes individually into a spectrophotometer, and record percent transmittance or absorbance in relation to a reagent blank. Data presented here demonstrate equivalent analytical performance of enzymatic reduction NO x methods in these various formats to that of benchmark CFA-CdR NO x methods.

Scientific Investigations Report↗

USGS2011

Colorimetric determination of nitrate plus nitrite in water by enzymatic reduction, automated discrete analyzer methods

This report documents work at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative methods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS automated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction methods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37°C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20°C to 37°C. These previously unrecognized AtNaR2 characteristics are essential for successful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here. Method detection levels (or limits; MDL) estimated for standard- and low-level DA-AtNaR2 nitrate + nitrite methods were 0.02 milligrams nitrogen per liter (mg-N/L) and 0.002 mg-N/L, respectively, which are comparable to 2010 NWQL long-term MDLs of the continuous-flow analyzer, cadmium-reduction methods (henceforth, CFA-CdR) they replace. Typically, reagent-water blanks for standard- and low-level DAAtNaR2 nitrate + nitrite methods are one half MDL or less. Nitrate + nitrite concentration differences for between-day replicates were 3 percent or less at or above 5 times the MDL and were as great as 35 percent near the MDL. Typically, nitrate spike recoveries from reagent water, surface water, groundwater, and high-phenolic-content, humic-acid-amended reagent water were 100±20 percent. In addition to operational details and performance benchmarks for these new DA-AtNaR2 nitrate + nitrite assays, this report also provides results of interference studies for common inorganic and organic matrix constituents at 1, 10, and 100 times their median concentrations in surface-water and groundwater samples submitted annually to the NWQL for nitrate + nitrite analyses. Paired t-test and Wilcoxon signed-rank statistical analyses of results determined by CFA-CdR methods and DA-AtNaR2 methods indicate that nitrate concentration differences between population means or sign ranks were either statistically equivalent to zero at the 95 percent confidence level (p ≥ 0.05) or analytically equivalent to zero-that is, when p < 0.05, concentration differences between population means or medians were less than MDLs.

Techniques and Methods↗

USGS2003

Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory-Evaluation of alkaline persulfate digestion as an alternative to Kjeldahl digestion for determination of total and dissolved nitrogen and phosphorus in water

Alkaline persulfate digestion was evaluated and validated as a more sensitive, accurate, and less toxic alternative to Kjeldahl digestion for routine determination of nitrogen and phosphorus in surface- and ground-water samples in a large-scale and geographically diverse study conducted by U.S. Geological Survey (USGS) between October 1, 2001, and September 30, 2002. Data for this study were obtained from about 2,100 surface- and ground-water samples that were analyzed for Kjeldahl nitrogen and Kjeldahl phosphorus in the course of routine operations at the USGS National Water Quality Laboratory (NWQL). These samples were analyzed independently for total nitrogen and total phosphorus using an alkaline persulfate digestion method developed by the NWQL Methods Research and Development Program. About half of these samples were collected during nominally high-flow (April-June) conditions and the other half were collected during nominally low-flow (August-September) conditions. The number of filtered and whole-water samples analyzed from each flow regime was about equal.By operational definition, Kjeldahl nitrogen (ammonium + organic nitrogen) and alkaline persulfate digestion total nitrogen (ammonium + nitrite + nitrate + organic nitrogen) are not equivalent. It was necessary, therefore, to reconcile this operational difference by subtracting nitrate + nitrite concentra-tions from alkaline persulfate dissolved and total nitrogen concentrations prior to graphical and statistical comparisons with dissolved and total Kjeldahl nitrogen concentrations. On the basis of two-population paired t-test statistics, the means of all nitrate-corrected alkaline persulfate nitrogen and Kjeldahl nitrogen concentrations (2,066 paired results) were significantly different from zero at the p = 0.05 level. Statistically, the means of Kjeldahl nitrogen concentrations were greater than those of nitrate-corrected alkaline persulfate nitrogen concentrations. Experimental evidence strongly suggests, however, that this apparent low bias resulted from nitrate interference in the Kjeldahl digestion method rather than low nitrogen recovery by the alkaline persulfate digestion method. Typically, differences between means of Kjeldahl nitrogen and nitrate-corrected alkaline persulfate nitrogen in low-nitrate concentration (< 0.1 milligram nitrate nitrogen per liter) subsets of filtered surface- and ground-water samples were statistically equivalent to zero at the p =level.Paired analytical results for dissolved and total phosphorus in Kjeldahl and alkaline persulfate digests were directly comparable because both digestion methods convert all forms of phosphorus in water samples to orthophosphate. On the basis of two-population paired t-test statistics, the means of all Kjeldahl phosphorus and alkaline persulfate phosphorus concentrations (2,093 paired results) were not significantly different from zero at the p = 0.05 level. For some subsets of these data, which were grouped according to water type and flow conditions at the time of sample collection, differences between means of Kjeldahl phosphorus and alkaline persulfate phosphorus concentrations were not equivalent to zero at the p = 0.05 level. Differences between means of these subsets, however, were less than the method detection limit for phosphorus (0.007 milligram phosphorus per liter) by the alkaline persulfate digestion method, and were therefore analytically insignificant.This report provides details of the alkaline persulfate digestion procedure, interference studies, recovery of various nitrogen- and phosphorus-containing compounds, and other analytical figures of merit. The automated air-segmented continuous flow methods developed to determine nitrate and orthophosphate in the alkaline persulfate digests also are described. About 125 microliters of digested sample are required to determine nitrogen and phosphorus in parallel at a rate of about 100 samples per hour with less than 1-percent sample in

Water-Resources Investigations Report↗

USGS2000

Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory-Determination of ammonium plus organic nitrogen by a Kjeldahl digestion method and an automated photometric finish that includes digest cleanup by gas diffusion

The National Water Quality Laboratory (NWQL) determined ammonium plus organic nitrogen (Kjeldahl nitrogen) by using semiautomated, block digester methods for filtered and whole-water samples from 1986 until October 1, 1991. During that time, phosphorus was determined by a persulfate digestion method. In 1991, projected increases in demand for both tests by the U.S. Geological Survey?s National Water-Quality Assessment Program led the NWQL to develop and validate methods for determining both analytes in a common digest. This report describes a rapid and accurate method to determine Kjeldahl nitrogen. The batch, high-temperature (block digester), Hg (II)-catalyzed digestion step used in the new methods I-2515-91/4515-91 is similar to U.S. Geological Survey methods I-2552-85/4552-85 and U.S. Environmental Protection Agency method 351.2 except that sample and reagent volumes are halved. Prepared digests are desolvated at 220 degrees Celsius (oC) and digested at 370oC in separate block digesters set at these temperatures, rather than in a single, temperatureprogrammed block digester. This approach permits 40 calibrants, reference materials, and samples to be digested and resolvated in about an hour. Ammonium ions originally present in samples, along with those released during the digestion step, are determined photometrically by an automated, salicylatehypochlorite Berthelot reaction procedure at a rate of 90 tests per hour. About 100 microliters of digest are required per determination. The upper concentration level is 10 milligrams per liter (mg/L) with a method detection level of 0.05 mg/L. Repeatability for a sample containing about 4.1 mg/L of Kjeldahl nitrogen in a high suspended-solids matrix is 3.1 percent. Between-day precision for the same sample is 4.8 percent. A gas diffusion cell in the air-segmented continuous flow analyzer eliminates particulates and ions that otherwise would interfere in the photometric finish. A singlechannel analyzer can process the resolvated digests from two pairs of block digesters each hour. Statistical analysis of paired data for about 1,500 samples determined by U.S. Geological Survey methods I-2552-85/4552- 85 and I-2515-91/4515-91 during method validation revealed a median concentration difference between the former and the latter methods of about 0.1 mg-N/L. This result was expected because digestion blank concentrations (nearly equal to 0.1 mg/L) were not subtracted from concentrations reported by methods I-2552-85/4552-85. A 10-year record of National Water Quality Laboratory Kjeldahl nitrogen blind blank concentration data also supports a step-change decrease in Kjeldahl nitrogen concentrations of about 0.1 mg/L after methods I-2552-85/4552-85 were replaced by methods I-2515-91/4515-91 on October 1, 1991. Somewhat larger concentration differences between the two methods were observed for a subset of about 350 samples with nitrate plus nitrite concentrations greater than 1 mg-N/L.

Open-File Report↗

USGS1999

US Geological Survey nutrient preservation experiment: Experimental design, statistical analysis, and interpretation of analytical results

This report describes the experimental details and interprets results from a study conducted by the U.S. Geological Survey (USGS) in 1992 to assess the effect of different sample-processing treatments on the stability of eight nutrient species in samples of surface-, ground-, and municipal-supply water during storage at 4 degrees Celsius for about 30 days. Over a 7-week period, splits of filtered- and whole-water samples from 15 stations in the continental United States were preserved at collection sites with sulfuric acid (U.S. Environmental Protection Agency protocol), mercury (II) chloride (former U.S. Geological Survey protocol), and ASTM (American Society for Testing and Materials) Type I deionized water (control) and then shipped by overnight express to the USGS National Water Quality Laboratory (NWQL). At the NWQL, the eight nutrient species were determined in splits from each of the 15 stations, typically, within 24 hours of collection and at intervals of 3, 7, 14, 22, and 35 days thereafter. Ammonium, nitrate plus nitrite, nitrite, and orthophosphate were determined only in filtered-water splits. Kjeldahl nitrogen and phosphorus were determined in both filtered-water and whole-water splits. Data on which this report is based, including nutrient concentrations in synthetic reference samples determined concurrently with those in real samples, are extensive (greater than 20,000 determinations) and have been published separately. In addition to confirming the well-documented instability of nitrite in acidified samples, this study also demonstrates that when biota are removed from samples at collection sites by 0.45-micrometer membrane filtration, subsequent preservation with sulfuric acid or mercury (II) provides no statistically significant improvement in nutrient concentration stability during storage at 4 degrees Celsius for 30 days. Biocide preservation had no statistically significant effect on the 30-day stability of phosphorus concentrations in whole-water splits from any of the 15 stations, but did stabilize Kjeldahl nitrogen concentrations in whole-water splits from three data-collection stations where ammonium accounted for at least half of the measured Kjeldahl nitrogen.

Water-Resources Investigations Report↗

USGS1995

U.S. Geological Survey nutrient preservation experiment: Nutrient concentration data for surface-, ground-, and municipal-supply water samples and quality-assurance samples

This report is a compilation of analytical results from a study conducted at the U.S. Geological Survey, National Water Quality Laboratory (NWQL) in 1992 to assess the effectiveness of three field treatment protocols to stabilize nutrient concentra- tions in water samples stored for about 1 month at 4C. Field treatments tested were chilling, adjusting sample pH to less than 2 with sulfuric acid and chilling, and adding 52 milligrams of mercury (II) chloride per liter of sample and chilling. Field treatments of samples collected for determination of ammonium, nitrate plus nitrite, nitrite, dissolved Kjeldahl nitrogen, orthophosphate, and dissolved phosphorus included 0.45-micrometer membrane filtration. Only total Kjeldahl nitrogen and total phosphorus were determined in unfiltered samples. Data reported here pertain to water samples collected in April and May 1992 from 15 sites within the continental United States. Also included in this report are analytical results for nutrient concentrations in synthetic reference samples that were analyzed concurrently with real samples.

Open-File Report↗

USGS1992

Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory-Determination of the total phosphorus by a Kjeldahl digestion method and an automated colorimetric finish that includes dialysis

A method to determine total phosphorus (TP) in the same digests prepared for total Kjeldahl nitrogen (TKN) determinations is desribed. The batch, high-temperature (block digester), HG(II)-catalyzed digestion step is similar to U.S. Geological Survey methods I-2552-85/I-4552-85 and U.S. Environmental Protection Agency method 365.4 except that sample and reagent volumes are halved. Prepared digests are desolvated at 220 degrees Celsius and digested at 370 degrees Celsius in separate block digesters set at these temperatures, rather than in a single, temperature-programmed block digester. This approach is used in the method escribed here, which permits 40 calibrants, reference waters, and smaples to be digested and resolvated in about an hour. Orthophosphate ions originally present in samples, along with those released during the digestion step, are determined colorimetrically at a rate of 90 tests per hour by an automated version of the phosphoantimonylmolybdenum blue procedure. About 100 microliters of digest are required per determination. The upper concentration limit is 2 milligrams per liter (mg/L) with a method detection limt of 0.01 mg/L. Repeatability for a sample containing approximately 1.6 mg/L of TP in a high suspended-solids matrix is 0.7 percent. Between-day precision for the same sample is 5.0 percent. A dialyzer in the air-segmented continuous flow analyzer provides on-line digest cleanup, eliminated particulates that otherwise would interfere in the colorimetric finish. An single-channel analyzer can process the resolvated digests from two pairs of block digesters each hour. Paired t-test analysis of TP concentrations for approximately 1,600 samples determined by the new method (U.S. Geologial Survey methods I-2610-91 and I-4610-91) and the old method (U.S. Geological Survey methods I-2600-85 and I-4600-85) revealed positive bias in the former of 0.02 to 0.04 mg/L for surface-water samples in agreement with previous studies. Concentrations of total phosphorus detemrined for ground-water samples by both methods were statistically equivalent.

Open-File Report↗