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Standard Operating Procedures

CRDS Water Isotope Analysis - δD δ18O

Introduction

Dr DeSoto is a Picarro L2120i liquid water cavity ring-down spectroscopy (CRDS) laser. dD and d18O of water are derived from absorption of specific spectral lines in the near-infrared range. Water vapor is introduced into an optical measurement cavity with an effective path length of up to 20 kilometers. We have two other Picarro liquid water isotope analyzers, Gorky, a L1102i that also measures dD and d18O and Phoenix, a L2140i that measures dD, d18O, and d17O.

Currently, each injection takes approximately 8 minutes and 30 seconds. A full tray of 54 vials with 10 injections each, then, will take about 3 days and 5 hours.

Safety

Users use be careful with syringe needles and vials. If either of these items are no longer useful (i.e. broken), they must be discarded into the broken glass container. The vaporizer is 110 *C and the septum nut can be hot to the touch.

Quick Start

  1. Clean up previous run – Close Coordinator, empty the existing tray, throw caps in trash, discard water down sink, vials go in beaker in drying oven.
  2. Load new waters - load 200 uL of water into 300 uL fused insert vials
  3. Take notes - Write the tray description and any other notes in the notebook while loading waters into vials.
  4. Prepare autosampler - Put vials in tray, clean syringe, modify job, don’t change the septum yet
  5. Fill out the daily log (DrDeSoto_DailyLog.ods)
  6. Start Run – open Coordinator Launcher, select High Precision, Start Autosampler, click Change Septum, wait for first injection to complete
  7. Take more notes – write down time and date of run start, what you did with the syringe, septum etc.
  8. Change Septum – when coordinator indicates the first injection is complete, press “Septum Changed” when done
  9. File management – make sure a directory has been created on the server for these samples (e.g. S:\data\projects\...).
  10. Tray Description – create tray description file in newly created folder, use excel, save as yymmdd_TrayDescription.csv
  11. Data - use matlab and the gorky script to reduce data from DeSoto

Exhaustive Protocol

Clean up previous run

Close the Coordinator...be patient, it will close eventually. Remove vials from the previous run and take them to the wet lab. Open each vial, throw caps in trash, and discard the water down the sink. If the vials contained dirty or salty samples, they must be soaked in DI water. You will likely have to pipette DI water into each vial. Once vials are clean, gently place vials in beaker and put them into the drying oven.

Load new waters

Load 200 uL of water into 300 uL fused insert vials. Use the pipette, pipette tips, and trays in 303B drawers labeled ‘Laser’. Vials are located on the lab bench above these drawers. Should you wish to load 1 mL of water, old vials are located in a drawer labeled ‘Laser – extras’. Use one full tray of dry clean pipette tips and also have one empty pipette tray available. For each distinct water, use a clean dry pipette tip, discarding the wet tips into the tray that started out empty. In this way, you will have one pipette tip tray with dry clean tips and one tray to receive the wet tips. If the pipette itself becomes wet, either on the inside or outside, ensure it is dry before proceeding. Use wipes or paper towels and / or compressed air to dry it out. Make sure the caps are just tight enough but not too tight. If the cap septum is overly concave, it is too tight. Put all standard waters and all sample waters back in the refrigerator when loading is complete. Put the wet pipette tip tray in the drying oven. As you are loading waters, write the water type or sample ID in the notebook.  

Take notes

Write the date, your name, sample set id, and the complete tray description as well as any other notes you desire in the notebook (Dr DeSoto) while loading waters into vials.

Prepare autosampler

Place the tray of samples and standards onto the tray-holder into either the rear or front position. Make certain the tray is properly seated in one of these two positions. Visually check the caps one last time; no cap should be overly concave.

DrDeSoto syringe cleaning
Using the Helper Syringe to apply 1% HCl to top of to-be-cleaned syringe.

Clean the syringe. Press “Menu” (F1), on autosampler controller, select "Change Syr" (also F1) (NOT "Clean Syr" - this is an automated method that we do not use). Remove the syringe from the autosampler by unscrewing the top red nut holding the plunger in place and by rotating the black syringe-body lock to a vertical position. Find the syringe cleaning kit. Fill the 'helper' syringe that is in the kit (not the one you just removed) with the 1% HCl solution and set aside. Place the syringe you just removed from the autosampler into the HCl solution and pull the plunger up half way or as far as you can. Use the 'helper' syringe and apply HCl to the top of the autosampler syringe (e.g. where the plunger meets the syringe glass body). Set the helper syringe aside. Continue moving the autosampler syringe up and down while it is in the HCl solution until plunger can be removed from syringe body (this usually takes a while and you must be patient and gentle). Wipe off plunger with a wipe. Reinsert the plunger into the body and continue moving the plunger up and down while still in the HCl. Remove the plunger again and wipe it off. Inject all HCl back into HCl container. Fill the autosampler syringe with DI water by pulling plunger all the way to the top. Release this water into a wipe, and repeat ~5 times. Rinse heavily with the same DI while keeping syringe tip in the DI water. Reinsert the, now clean, syringe into the autosampler and indicate complete on autosampler controller. Rinse the 'helper' syringe with DI water in the same way. If you can’t remove the plunger during cleaning or break it or it was already broken, obtain a new 10 uL syringe from drawer under Dr DeSoto. Discard the old syringe to the broken glass box in 303B.

Modify the autosampler job according to your current number of vials and your preferred number of injections per vial. From the home screen on autosampler controller, turn the wheel to highlight the appropriate job and center click. 'First' should be 1, 'Last' should be the last vial in your tray (e.g. 54), 'Increment' should be 1, 'Count' should be the number of injections per vial you wish to have (e.g. 10). To change any of these values, rotate the wheel until the value is highlighted, center click, and turn the wheel to edit the number. Center click again when you are done editing. Press 'Home' (the F4 button) when you are finished editing the job.

Daily Log

Fill out all information within the file ‘DrDeSoto_DailyLog.ods’. Make sure you note the H2O ppm background before before starting the run.

Start Run

Open the Coordinator Launcher, select High Precision or High Throughput, Start Autosampler (F4 from Home screen). Assuming you are running only one tray, rotate wheel until 'selected' is highlighted and center click. If you have two trays, select 'All' with a center click. Click "Change Septum" (upper right corner of Coordinator window) and wait for first injection to complete.

Tray Description

While waiting for Dr DeSoto to be ready to have its septum changed, type in tray description, use C:\DrDeSoto\TrayDescriptions\_Rear_TrayDescription.csv (or *\_Front_* if you are loading a tray for the front position) and save it as yymmdd_TrayDescription.txt.

Load Sample Description by clicking ‘Load Sample Descriptions’ and then browse to the tray description you just typed in (c:\DrDeSoto\).

Change Vaporizer Septum

DrDeSoto septum change
The swage-lok nut, large gauge needle, and vaporizer septum.

After the first injection is complete the coordinator will indicate when its appropriate to change the septum. Remove the magnetic cover and use the slotted wrench side of this cover to loosen the nut. Remove the nut with your fingers taking care as it is hot. The septum should be stuck inside the nut. Use the large-gauge needle that is in the syringe cleaning kit to extract the septum. Discard the old septum into the trash. Place the new septum into swage-lok nut using the blunt end of large-gauge needle. Place the nut back onto the vaporizer taking care not the allow the septum to fall out when you turn the nut over. Hand tighten the nut; you should feel the nut bottom out. Press “Septum Changed” which is the same button you pushed to begin changing septum in Coordinator window.

Take more notes

Write down the time and date of run start. This is essential as files are named within the instrument based on time. Also write down what you did with the syringe, the septum, etc.

Data

Make sure a directory on the server has been created (e.g. S:\Data\projects\amazingscience\) and then create ‘raw’, ‘reduced’, and 'traydescriptions' folders within there. Copy the data from C:\IsotopeData\ to the 'raw' folder. Your file will have the date and time you started it as part of the filename (e.g. HBDS2213_IsoWater_20160223_151837.csv) where HBDS2213 is the instrument serial number and the time is so lovingly put in GMT, thanks Picarro. Also copy the TrayDescription file you created on DeSoto to the 'TrayDescription' folder.

Type 'gorky' into matlab command prompt. Browse to raw file you just transferred from DeSoto and open it / select it. Follow prompts in matlab. In the end, a reduced txt file is created 

Troubleshooting