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[[Nitrate]], the most oxidized form of nitrogen, is regulated to protect people and animals from harmful levels as there is a large over abundance due to anthropogenic factors. Widespread field testing for nitrate could begin to address the nitrate pollution problem, however, the Cadmium Reduction Method, the leading certified method to detect and quantify nitrate, demands the use of a toxic heavy metal. An alternative, the recently proposed [[Environmental Protection Agency]] Nitrate Reductase Nitrate-Nitrogen Analysis Method, eliminates this problem but requires an expensive proprietary spectrophotometer. The development of an inexpensive portable, handheld photometer will greatly expedite field nitrate analysis to combat pollution. To accomplish this goal, a methodology for the design, development, and technical validation of an improved open-source water testing platform capable of performing Nitrate Reductase Nitrate-Nitrogen Analysis Method. This approach is evaluated for its potential to i) eliminate the need for toxic chemicals in water testing for nitrate and nitrite, ii) reduce the cost of equipment to perform this method for measurement for water quality, and iii) make the method easier to carryout in the field. The device is able to perform as well as commercial proprietary systems for less than 15% of the cost for materials. This allows for greater access to the technology and the new, safer nitrate testing technique.
==Enzymes and Photometers Poster at Pittcon 2016 ==
* W.H. Campbell, E.R. Campbell, D.A. Squires, J. Walbeck, W. Ashbaugh, A. Davidson, B.T. Wittbrodt, J.M. Pearce. Enzymes and Photometers. ''This poster was presented at Pittcon 2016 - Session 820 - Portable Instruments'' [ download poster]
NECi Superior Enzymes develops wet chemistry methods that replace metals and other chemicals with recombinant protein reagents. These protein reagents are produced in tightly controlled protein expression systems for lot-to-lot consistency and therefore can be regarded as any other reagent-grade chemical. NECi’s nitrate reductase enzyme reagents for nitrate quantification have been on the commercial market for over ten years[3]. Method validations for the nitrate reductase method include USGS, ASTM, and pending validations for the U.S. EPA Clean Water Act+, Safe Drinking Water Act+ and Standard Methods+. NECi Superior Enzymes has since developed enzyme-based test methodologies for quantification of phosphate[2], and the research and development team is currently developing methods for the quantification of ethanol, glycerol, and galactose. The idea for this project was prompted by the lack of economical photometers compatible with standard 1ml cuvettes to provide quantitative data from the methods described above.
+Pending Methods
# Wittbrodt BT, Squires DA, Walbeck J, Campbell E, Campbell WH, Pearce JM (2015) Open-Source Photometric System for Enzymatic Nitrate Quantification. PLoS ONE 10(8): e0134989. doi:10.1371/journal.pone.0134989
# Campbell ER, Warsko K, Davidson A-M, Campbell, WH, (2015) Determination of phosphate in soil extracts in the field: A green chemistry enzymatic method. Methods X 2, pp211-218. doi:10.1016/j.mex.2015.04.003
# Campbell WH, Song P, Barbier GG (2006) Nitrate reductase for nitrate analysis in water. Environmental Chemistry Letters 4(2) pp 69-73. doi:10.1007/s10311-006-0035-4
==Bill of Materials==

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