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Measuring N and O isotopes in clean green New Zealands water: method adaption. National Isotope Centre GNS Science Andy Phillips The Methods available. 1) Ion exchange column collection 2) Microbial bacteria method 3) Enzyme

  1. Measuring N and O isotopes in clean green New Zealand’s water: method adaption. National Isotope Centre GNS Science Andy Phillips

  2. The Methods available. • 1) Ion exchange column collection • 2) Microbial bacteria method • 3) Enzyme method • 4) Cadmium reduction GNS Science GNS Stable Isotope Laboratory

  3. Cadmium Method 1 st Step • 1 ST Attempt • Making Spongy Cadmium using Zinc Sticks • Sample preseravtion NaOH pellets • Imadazole as sample Buffer • 2 nd Attempt • Cadmium powder HCl wash • Sample preseravation HCl pH 2-3 • Addition of Sulfanilic Acid to remove Nitrite GNS Science GNS Stable Isotope Laboratory

  4. Nitrate to Nitrite by Cadmium reduction • This takes place in 6M NaCl, the excess Cl ions • restricts oxygen exchange from the water. • NaCl pre-baked at 700C for 2 hrs • MgO pre-Baked at 450C for 1 hr • Cd powder 0.7g per sample has10% HCl added to it the clumps that form are chomped up to form a granular consistancy and then rinsed 10 times with DIW and stored under it until ready to use. GNS Science GNS Stable Isotope Laboratory

  5. • Sample was added to 6M sodium chloride solution,100mg of MgO and 7g of spongy Cadmium in a 50mL centrifuge tube, giving a total volume of 20mL. • The bottle was shaken horizontally for 17 hours for the reduction to take place. • Centrifuge to trap all the Cadmium at the base of the tube. GNS Science GNS Stable Isotope Laboratory

  6. Day2 Azide Reaction • HNO2 + HN3------N2O + H2O + N2 • Azide solution was added to Acetic Acid to give a 1:1 mix and sparged with He for 30mins • 450uL Azide mix is injected in sealed 12mL capped vials containing sample from the Cd step plus 2M NaCl • The vials are left for 30mins at room temperature • To stop the reaction 450uL of 6M NaOH is injected into the vial GNS Science GNS Stable Isotope Laboratory

  7. Cd Step ( m L) Azide Step ( µL) Sample Name Sample Info Nitrate (ppm) mL sample 6M NaCl 100 m g MgO 0.7 g Cd DI w ater Final NO2- Conc Sam ple from Cd A:A 2M NaCl Final ppb N2O nm ol 6.7 1 BLANK No Cadmium step 0.012 13.3 100 0.0 0.01 7500 450 50 7.5 4.3 6.7 2 BLANK Cd 0.001 13.3 100 0.70 0.0 0.00 7500 450 50 0.6 0.4 6.7 3 USGS 34 a 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 6.7 4 I AEA a 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 6.7 5 KNO3 a 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 6 KNO3 c from b Repeat Azide step 1.000 0.35 200 450 7350 8.8 5.0 7 KNO3 d from b Repeat Azide step 1.000 0.35 80 450 7470 3.5 2.0 6.7 8 KNO3 e 0.2 ppm 0.200 13.3 100 0.70 0.0 0.13 840 450 6700 14.0 8.0 6.7 9 KNO3 f 0.05 ppm 0.050 13.3 100 0.70 0.0 0.03 3360 450 4190 14.0 8.0 6.7 10 KNO3 g 0.02 ppm 0.020 13.3 100 0.70 0.0 0.01 7500 450 50 12.5 7.1 6.7 11 MAA1 a No Cadmium step 1.000 7.0 100 6.3 0.35 320 450 7230 14.0 8.0 6.7 12 MAA2 a No Cadmium step 1.000 7.0 100 6.3 0.35 320 450 7230 14.0 8.0 6.7 13 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 14 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 15 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 16 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 17 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 18 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 19 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 20 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 21 USGS 34 c from a Repeat Azide step 1.000 0.35 320 450 7230 14.0 8.0 6.7 22 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 23 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 24 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 25 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 26 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 27 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 28 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 29 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 30 USGS 34 d from a Repeat Azide step 1.000 0.35 320 450 7230 14.0 8.0 6.7 31 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 32 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 33 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 34 0 0.000 # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 35 0 0.000 # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 36 0 0.000 # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 37 Dup (high NO3) # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 38 Dup (low NO3) # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 39 Dup (med NO3) # DI V/ 0! 100 0.70 # # # # # DI V/ 0! # DI V/ 0! 450 # DI V/ 0! # DI V/ 0! # # # 6.7 40 USGS 34 b Dup 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 6.7 41 I AEA b Dup 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 6.7 42 KNO3 b Dup 1.000 7.0 100 0.70 6.3 0.35 320 450 7230 14.0 8.0 43 MAA1 b from a Repeat Azide step 1.000 0.35 320 450 7230 14.0 8.0 44 MAA2 b from a Repeat Azide step 1.000 0.35 320 450 7230 14.0 8.0 45 Tester (USGS34 a) Repeat Azide step 1.000 0.35 320 450 7230 14.0 8.0 240.5 23.1 #DIV/0! GNS Science GNS Stable Isotope Laboratory

  8. d15N d18O Stdev d15N Stdev d18O USGS 34 -5.6 -24.9 0.1 0.5 IAEA -1.7 21.6 0.0 0.3 KNO3 0.4 10.1 0.0 0.1 USGS 34 c -5.6 -25.1 0.1 0.4 IAEA c -1.7 21.7 0.1 0.2 KNO3 h 0.4 9.9 0.1 0.3 USGS 34 b1-1 h -5.4 -24.0 0.3 0.1 USGS 34 b1-1 h -5.6 -24.8 0.1 0.1 USGS 34 b1-30 min -5.6 -24.8 0.1 0.1 USGS 34 b1-10 min -6.20 -22.34 0.4 0.7 Average 0.1 0.2 GNS Science GNS Stable Isotope Laboratory

  9. Results SAMPLES nA d15N d18O d15N corr for 17O d15N known d18O known d15N line d18O line SLOPE 0.43 0.87 INTERCEPT -4.46 -0.44 11.05 -5.66 USGS 34 -25.2 -5.27 -1.8 -27.9 -1.9 -28.4 9.64 -1.76 IAEA-NO3 21.4 -2.42 4.7 25.6 4.7 25.1 9.93 0.40 KNO3 10.1 0.15 10.7 11.73 10.7 12.0 11.08 -5.58 USGS 34 -24.5 -5.21 -1.8 -27.9 -1.7 -27.6 10.20 -1.72 IAEA-NO3 21.9 -2.39 4.7 25.6 4.8 25.6 9.07 0.39 KNO3 10.2 0.14 10.7 11.73 10.7 12.2 9.74 -29.17 MAA1 -3.1 -30.59 -60.6 -60.7 -3.1 10.32 -5.08 MAA2 -2.6 -5.27 -3.9 -1.9 -2.5 9.60 -0.90 Tap Water 1.8 -0.99 8.0 2.5 4.61 -5.60 Ice Core Water a -3.8 -5.79 -3.1 -3.9 4.47 -6.24 Ice Core Water b -6.4 -6.39 -4.5 -6.8 USGS 34 c -5.6 -25.1 -5.24 -1.8 -28.3 IAEA c -1.7 21.7 -2.32 4.9 25.4 KNO3 h 0.4 10.1 0.12 10.6 12.0 USGS 34 b1-10 min -6.20 -22.34 -5.92 -3.4 -25.1 USGS 34 b1-1 h -5.4 -24.0 -5.04 -1.4 -27.0 USGS 34 b1-1 h -5.6 -24.5 -5.20 -1.7 -27.5 USGS 34 b1-30 min -5.6 -24.8 -5.18 -1.7 -27.9 GNS Science GNS Stable Isotope Laboratory

  10. GNS Science GNS Stable Isotope Laboratory

  11. GNS Science GNS Stable Isotope Laboratory

  12. GNS Science GNS Stable Isotope Laboratory

  13. GNS Science GNS Stable Isotope Laboratory

  14. GNS Science GNS Stable Isotope Laboratory

  15. GNS Science GNS Stable Isotope Laboratory

  16. GNS Science GNS Stable Isotope Laboratory

  17. GNS Science GNS Stable Isotope Laboratory

  18. GNS Science GNS Stable Isotope Laboratory

  19. Ammonium • Using method by Zhang and Altabet • Samples need to have had Sulfanilic Acid added • 2mL 6M NaCl and 0.8mL Hypobromate + sample • 30 mins RT then inject Arsenite to nullify the Hypobromite. • Caps are screwed on and the Azide is added. GNS Science GNS Stable Isotope Laboratory

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