SOP Field sampling for nitrous oxide (N 2 O), carbon dioxide (CO 2 ), and methane (CH 4 ) fluxes. Procedure Overview

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1 SOP Field sampling for nitrous oxide (N 2 O), carbon dioxide (CO 2 ), and methane (CH 4 ) fluxes Procedure Overview These greenhouse gases are emitted from soils at various rates dependent on environmental conditions, vegetation on site, fertilization regimes, among other factors. The following procedure is designed to collect samples of the emitted gases over time to determine gas flux. Collected samples are later analyzed on a gas chromatograph (GC) with flame ionization (FID) and electron capture detectors (ECD) to quantify each greenhouse gas. Equipment: Chamber Top Assembly: PVC pipe, 11 7/8 inner diameter, schedule 40; cut this into 2 pieces. One piece will be 3 7/8 tall, with a 3/8 hole in the side (bottom of hole will be 1 from the top of the PVC edge). This will be the chamber top. The second piece will be 5 tall, with a ~80º bevel put on last 3/4 of bottom edge. This will be the sampling ring that goes into the ground. Gray PVC sheet, ¼ thick, cut into a 12 ¾ circle, with (2) ½ holes drilled equidistantly from each other in the center. Note: Both the PVC pipe and the sheet plastic will need to be cut and milled by an experienced shop with appropriate machinery Chemistry Machine Shop constructed all the PVC pieces used by our group in the past. Once milled, the PVC portion of the chamber tops and rings should last a long time; only the reflective tape and seals will need to be replaced from time to time. Epoxy glue Plastic joint (Cole-Parmer Instrument Co., # PK, straight ¼ union) Stainless steel tubing: 4.8 mm diameter by 10 cm length. Order 3 ft. lengths from Cole- Parmer (#EW Stainless Steel Tubing; dimensions, 3/16" ID, 1/4" OD, 1/32" wall) and cut to size using tubing cutter). Tractor tire inner tube (15.5R38 check area tire shops to see if they have any leaky ones; for this purpose, you can work around any holes. If not, you ll have to buy one new). Duct tape Reflective tape (Shurtape Hold Strong available at Lowe s) Rubber weather stripping with adhesive Field Sampling: Glass serum vials, 6.0 ml (22*38mm), Alltech #98768 Rubber septa (Voigt Global 20mm legless vial stoppers - item #73828A-RB (lot # worked well for us). Riparian Buffer Group p. 1

2 20mm aluminum crimp top seals (Fisher Scientific # C) Seal crimper Evacuation manifold Helium gas for the manifold; % purity, 244 ft 3 (from Chemistry Stores item # A) Digital thermometer (from Cole-Parmer; Low-Cost Waterproof Pocket Thermometers, item #EW ) A reflective cup pierced with multiple holes to be attached to a digital thermometer (a film canister or other similarly shaped plastic cylinder covered with reflective tape works well). Portable soil moisture sensor (we use the Campbell Scientific Hydrosense ) or prepare to take gravimetric soil moisture samples) 10 ml disposable plastic syringes 22 gauge, 5/8 inch long syringe needles Garden hand clippers Timepiece (to keep track of minutes elapsed a cell phone clock will work) Centimeter ruler Clipboards Data sheets (may want to print these on Rite-in the-rain paper) Pens/pencils Assembly of Chamber Top (Figures 1 and 2): 1. Epoxy the unbeveled PVC ring of the chamber top to the circular plates forming the cap of the gas chamber. 2. Drill two holes through the top plate and one threaded port through the side of the top to accept the vent and tubing (if this hasn t already been done). 3. Attach ¼ inch stainless steel tubing to plastic vent and thread into the side of the chamber cap. 4. Cut the tractor inner tube into rings ~7cm wide and stretch over the chamber cap such that half of the tubing hangs free. This will be stretched over the bottom ring of the chamber forming a seal between the two. Duct tape inner tubing to the chamber cap. 5. Apply reflective tape to outside top and sides of the chamber cap to prevent absorption of solar radiation. 6. Apply weather stripping to the bottom edge of the chamber cap to help form a tight seal between the cap and the bottom ring. Riparian Buffer Group p. 2

3 7. Place one septum into one of the holes in the chamber cap and a thermometer pierced through a septum into the other hole. Figure 1. Left photo: The chamber cap located is made up of the top plate (gray) and the bottom PVC ring. Note the vent hole drilled into the PVC in the front. Right photo: Inside of a finished cap complete with vent tubing, weather stripping, and inner tube seal (folded back, ready for deployment). Note that the weather stripping on this cap is loose and should be replaced. Riparian Buffer Group p. 3

4 Figure 2. View of the gas chamber cap attached to the bottom ring. Two port holes on top are fitted with rubber septa, one of which holds a digital thermometer. A digital thermometer is also shown with a reflective cup for the ambient air temperature measurement. Procedure: 1. Install chamber rings into the desired location a few days prior to sampling. Use a mallet and a board to drive the ring into the soil until 6 or 7 cm remain exposed. 2. Before sampling, prepare glass vials by crimping the septa and aluminum rings tight to each vial using the handheld crimper. 3. Each plot to be sampled will require 4 vials for each chamber, plus an additional vial per plot for a ambient air sample; # vials = (# rings * 4) + 1. Label these vials to reflect the 0, 15, 30, and 45 minute samples taken from each ring (1-4, or A-D). 4. Using the evacuation manifold and helium (set the helium regulator to about psi), alternate evacuating and flushing with helium on 20 second intervals until the vials have Riparian Buffer Group p. 4

5 been evacuated 5 times. Extend the final evacuation to 90 seconds. Close port valves before turning off the evacuation pump to prevent back-flush into the vials. 5. To begin sampling in the field, clip all living vegetation to the soil surface (DO NOT pull vegetation as this will disturb the soil and affect results) inside and immediately outside the chamber ring installed in the field plot in step Measure the inside height (anchor length in cm) of the ring in 3 places. The average of these will be used to calculate the internal volume of the chamber when added to the known volume of the chamber cap. 7. Place the chamber cap over the ring, taking care to point the chamber vent away from the prevailing wind. 8. Fold down the free half of the inner tubing to create a tight seal between the cap and bottom ring - this is most easily done by kneeling on the cap. Make sure no vegetation is caught between the chamber top and bottom, or within the inner tube seal, as this will affect the gas-tight seal needed. 9. Install septa and septa with thermometer into the holes in the chamber top. Note: Keep thermometers shaded prior to this to prevent false initial temperature readings. 10. Immediately after the chamber is sealed and thermometer and septa are installed, purge the 10 ml syringe by pumping it 3 times into the air, downwind. Draw a full syringe (go past 10 ml it is easier to feel this happen than it is to read the 10 ml mark on the syringe) of sample from the gas chamber through the septum on the cap not pierced by the thermometer. Inject this sample into the corresponding vial. 11. Record the internal temperature of the chamber at the time the chamber gas was sampled using the thermometer passing through the septum on the chamber cap. 12. Continue taking samples after 15, 30 and 45 minutes and recording chamber temperatures at each of these times. (DO NOT remove chamber cap, septum, or thermometer until the 45 minute sample has been taken). Note: While waiting to resample this chamber, others can be set up and sampled to save time (approximately 5 or 6 chambers can set up to be sampled at one time, provided the chambers are within a reasonable walking distance). Additionally, steps can be completed at any time while waiting for the 45 minutes to elapse. 13. Attach an available thermometer to the reflective cup to record the air temperature for the plot. Use the reflective cup to prevent direct sunlight from hitting the tip of the thermometer (this can take several minutes). Riparian Buffer Group p. 5

6 14. Take a sample of the ambient air by purging the syringe 3 times (as if to take a chamber air sample), and pull a full syringe of air from downwind. Place in the appropriate sample vial. 15. Record soil temperature at each gas chamber at 5 and 10 cm by pushing an available digital thermometer into the soil outside each chamber. (Insert thermometer half way for 5 cm reading and all the way for the 10 cm reading). 16. It is easiest to remove the chamber cap from the ring by again kneeling on the cap and folding up the inner tubing. Rings should be left in field plots for future sampling, unless they conflict with field management. You have now finished your first gas sampling experience! 17. Gas sample vials should be stored at room temperature until ready for analysis on the GC. Written By: Eric Kilburg Date: 28 July 2010 Approved By: Leigh Ann Long Date: 9 August 2010 Riparian Buffer Group p. 6

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