1. iso TOC cube: new innovations from a familiar company Advances in Stable Isotope Techniques & Applications, University of Calgary Tuesday June 4 th, 2013 Arthur Kasson, IRMS Product Manager elementar Americas Inc. 1

2. Compact, bench-top IRMS 100% high purity stainless steel horizontal vacuum housing Extremely high vacuum conductance means only a single turbomolecular pump is required 100V dynamic range linear analyzer Robust thorium coated ionization filament 10 year warranty of Faraday Collectors Modular electronics for simple maintenance High stability electromagnet with no need for water cooling Fully automated analysis and diagnostics through IonVantage software Market leading performance specifications Compact, bench-top IRMS 100% high purity stainless steel horizontal vacuum housing Extremely high vacuum conductance means only a single turbomolecular pump is required 100V dynamic range linear analyzer Robust thorium coated ionization filament 10 year warranty of Faraday Collectors Modular electronics for simple maintenance High stability electromagnet with no need for water cooling Fully automated analysis and diagnostics through IonVantage software Market leading performance specifications 2

3. Sensitivity Absolute sensitivity of CO 2 (molecules/ion) 850 DI mode 1200 CF mode H 3 + correction factor< 8.0 ppm/nA H 3 + factor stability< 0.03 ppm/nA/hr Mass Resolution (10% valley definition)100 GasIsotope Internal Precision 1σ ‰ Linearity ‰ / nA CO 2 δ 13 C δ 18 O ≤ 0.06 ≤ 0.02 ≤ 0.04 N2N2 δ 15 N≤ 0.06≤ 0.02 H2H2 δDδD≤ 0.20- IRMS Reference Gas IsoPrime100 Specifications 3

4. The main applications for TOC are environmental forensics and ecology 1. River nutrient and food webs 2. Agricultural run off into water systems 3. Soil dynamics 4. Water quality analysis and pollution 5. Marine & estuarine dynamics Total Organic carbon (TOC) IRMS Key Applications 4

5. 1. Variable sample feeding 2. High temperature digestion with matrix separation (up to 1200C) 3. Highly stable operation and results 4. NDIR detection of TOC in the ppb to percentage range (can also measure TN b ) 5. Ability to remove excess salt via ash crucible Total Organic carbon (TOC) IRMS Unique Capabilities 5

6. Preparation off-line Large sample sizes, complicated methodology, specialised equipment, analysis by dual inlet, slow Iso TOC cube Current Techniques Wet oxidation TOC-IRMS Using UV or persulfate for oxidation of the sample. Few samples can be analysed before halide gasses corrode reaction vessel, salt deposition in flow lines, consumption of halogen trap EA-IRMS Concentration by evaporation or lyophilisation, analysis by EA-IRMS Does not work well when high concentrations of inorganic salt are present 6

7. Elementar have over 30 years of experience in manufacturing TOC analysers The vario TOC cube is the latest version in the cube format Iso TOC Cube 7

8. Integrated 50 position liquid sampler with integrated automated sample feeder Iso TOC Cube Features 8

9. Screwless casing with easy access to 5 sides of the instrument Simple maintenance with immediate access to all parts Features Iso TOC Cube 9

10. Furnace Syringe pump Acid reservoir Drying tube Halogen trap Multiport valve Features Iso TOC Cube 10

11. Use of ball & clamp fittings throughout the instrument for simple maintenance with no need for tools Ball & Clamp Fittings Iso TOC Cube 11

12. Requires interface for IRMS for coupling to IsoPrime100Requires interface for IRMS for coupling to IsoPrime100 Benefits during IRMS Iso TOC Cube Iso TOC cube uses direct analysis of TIC & TOCIso TOC cube uses direct analysis of TIC & TOC Does not use subtraction method (TC - TIC = TOC) High temperature oxidation furnace for combustion of C to CO2High temperature oxidation furnace for combustion of C to CO2 Does not use wet oxidation (persulfate/UV oxidation) O2 carrier gas usedO2 carrier gas used Does not use synthetic air Combustion tube containing CuO @ approx 950 °CCombustion tube containing CuO @ approx 950 °C Does not use expensive platinum 12

13. IRMS Interface – Trap TIC/TOC CO 2 13

14. IRMS Interface Introduction Iso TOC Cube 14

15. CO2 Column Trapping Iso TOC Cube 15

16. IRMS Interface – Release Trapped CO 2 16

17. CO2 Column Release to the IRMS Iso TOC Cube 17

18. 18 System configuration Exchange from O 2 to He carrier gas Purge and trap adsorption column for peak focusing (no liquid nitrogen needed) reduction furnace to remove interference species and residual oxygen

19. 13/05/2011 (7.33 to 10.24) TOC-IRMS Settings C-concentration ~ 1ppm Sample volume ~ 3.5ml CO2 Adsorption by Silica gel Trap 550 uA IR bridged (1/16th" Haler) Peak Ave time N = 285 He Purge = 30s SamplenAδ 13 C CitricAcid15.30-18.04 CitricAcid24.65-17.27 CitricAcid34.60-17.23 CitricAcid44.37-17.28 CitricAcid54.25-17.48 CitricAcid64.48-17.15 CitricAcid74.53-17.24 CitricAcid84.43-17.24 CitricAcid94.38-17.24 CitricAcid104.46-17.23 Ave-17.26 SD0.089 ~1ppm C Precision Iso TOC Cube 19

20. 13/05/2011 (10.41 to 13.15) TOC-IRMS Settings C-concentration ~ 0.15ppm Sample volume ~ 3.5ml CO2 Adsorption by Silica gel Trap 550 uA IR bridged (1/16th" Haler) Peak Ave time N = 285 He Purge = 30s SamplenAδ 13 C DI water 50.35-33.70 DI water 60.36-34.19 DI water 70.32-34.06 DI water 80.37-34.64 DI water 90.33-34.54 DI water 100.32-34.74 Ave-34.31 SD0.40 ~0.15ppm Water Iso TOC Cube 20

21. 10/05/2011 (7.38 to 16.12) TOC-IRMS Settings C-concentration ~ 10ppm sample vol ~ 1.5ml CO2 Adsorption by Silica gel Trap 200 uA IR bridged (1/16th" Haler) Peak Ave time N = 285 He Purge = 30s SamplenAδ 13 cAvestd devActual ANU Sucrose 16.33-11.65 ANU Sucrose 26.29-10.72 ANU Sucrose 36.35-10.57 ANU Sucrose 46.44-10.52 -10.550.04-10.50 Citric Acid 16.48-16.18 Citric Acid 26.47-16.24 Citric Acid 36.4-16.17 Citric Acid 46.35-16.10 -16.170.06-16.00 RossmSugarA16.35-15.92 RossmSugarA26.18-15.90 RossmSugarA36.26-15.94 RossmSugarA46.13-15.90 -15.920.02-16.30 Glutamic AcidA16.28-26.19 Glutamic AcidA26.16-26.21 Glutamic AcidA36.13-26.11 Glutamic AcidA46.04-26.21 -26.180.05-26.10 Glutamic AcidB15.95-27.89 Glutamic AcidB25.98-27.92 Glutamic AcidB35.91-27.95 Glutamic AcidB46.29-27.98 -27.940.04-28.40 Testsugar16.02-25.57 Testsugar26.11-25.55 Testsugar36.01-25.56 Testsugar46.13-25.56 0.01-26.00 RossmSugarB15.98-25.16 RossmSugarB25.98-25.18 RossmSugarB35.99-25.13 RossmSugarB45.96-25.13 -25.150.02-25.70 ANU Sucrose 55.96-10.80 ANU Sucrose 65.93-10.57 ANU Sucrose 75.89-10.53 ANU Sucrose 85.88-10.54 -10.54670.02-10.50 TOC-IRMS Accuracy Iso TOC Cube 21

22. TOC-IRMS Accuracy Iso TOC Cube 22

23. 23 Stability (precision) and memory effect Absolute shifts between 11-39‰ Low C (5-12nA signal) Absolute shifts between 6-73‰ High C (16-55nA signal) Very good precision: ≤ 0.20‰ and ≤ 0.1‰ for shift below 30‰ Still very good precision: ≤ 0.20‰ (shift larger than 60‰ ≤ 0.30‰) Signal Absolute shift Injection 2,3,4  13 C stdev Injection 3,4,5  13 C stdev [nA][‰] 511.140.050.04 530.100.030.04 538.620.190.10 537.700.150.07 1223.080.02 1237.560.060.04 1218.430.100.04 1210.920.050.03 1230.340.090.04 1237.790.070.05 1214.550.090.02 1223.000.100.05 1638.870.130.08 1617.510.090.10 1615.650.180.02 5573.070.270.14 5562.300.230.13 5515.410.05 5516.100.130.02 5511.160.040.06 555.610.040.02 With 5 injections all data ≤ 0.10‰ (shift larger than 60‰ ≤ 0.20‰)

24. 24 Accuracy All data corrected with 2 points calibration curve Caffeine (IAEA-600)  13 C Glucose (IAEA-CH6)  13 C -27.77‰ -10.45‰ Average difference between measured and target value Only in 3 over 13 measurements differences ≥ 0.15‰ y = 1.00031x + 0.04485 r 2 = 0.9999 0.09‰

25. 25 Linearity of the system and concentration range (data courtesy of Dr. Chiara Cerli – U. Amsterdam) except for L-trypthophane (0.07 ‰/nA) humic acid (0.06 ‰/nA) glucose (0.04 ‰/nA) Between 5-150ppm (5-70nA) Very good linearity: ≤ 0.03 ‰/nA Between 1-150ppm (1-70nA) Very good linearity: ≤ 0.03 ‰/nA

26. 26 Real samples Rice (std dev 0.05‰) Moss (std dev 0.17‰) Forest floor (std dev 0.15‰) Peat (std dev 0.03‰) H horizon from Podzol (std dev 0.02‰) DOM extracted from: 0.5 ml injection, different concentrations Measured very well

27.  13 C 10-150 mgC L -1 stdev10-100 mgC L -1 stdev [‰] Citric acid-25.350.18-25.440.11 Benzoic acid-28.920.34-28.910.40 L-Trypthophane-11.350.31-11.470.09 Acetovanillone-30.530.15-30.610.07 Acetovanillone (fix C)-30.580.03-30.570.03 Urea-50.050.17-50.130.12 Caffeine-49.060.23-49.160.18 Mixture-28.930.62-29.080.68 Humic acid-26.180.30-26.210.35 Humic acid (fix C)-26.340.03-26.350.03 Melamine-19.770.23-19.890.15 Glucose-11.160.21-11.240.19 DOM_Mossnd -32.700.17 DOM_Moss (fix C)-32.810.09-32.800.09 27 Constant C injection (25mgC, ~22nA) Inj vol 0.08-2.5ml Very good std dev: ≤ 0.09 ‰ Constant inj vol (0.5ml) Various C concentrations (10-150 mg L -1 ) Good std dev: ≤ 0.30 ‰ Real samples: concentration vs volume (data courtesy of Dr. Chiara Cerli – U. Amsterdam)

28. 28 Conclusions stable system (large ‰ shift) accurate Wide range of system linearity (2-80nA <0.03‰/nA) flexibility for handling real samples Injection volumes (0.05-4ml) 1 nA/µg C Performances very satisfying `high throughput, easy to use and maintain Very interesting tool that widen opportunities for studies in environmental research

29. Special thanks go out to Dr. Chiara Cerli at the University of Amsterdam (Institute for Biodiversity and Ecosystem Dynamics) for providing us with additional data. 29 Acknowledgements Paul Wheeler, Mike Seed, Will Price, Rob Berstan Federherr E., H.P. Sieper, Lutz Lange, H.J. Kupka, R. Dunsbach, F. Volders

30. Schematic Outline of the Iso TOC Cube INTRODUCTION 30

31. Sample vial Sparger Furnace Condenser Sample Syringe Halogen trap Nafion Membrane Drying tube IR detector 31

32. Acidify Sparge Vessel to remove TIC STEP ONE 32

33. 33

34. Flush sample lines to waste and fill syringe STEP TWO 34

35. 35

36. Dose sample for TIC removal STEP THREE 36

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38. Measure TIC STEP FOUR 38

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40. Measure TOC STEP FIVE 40

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