CRUDE OIL AND OIL DISPERSANT EXPOSURE EXPERIMENT PROTOCOLS I

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CRUDE OIL AND OIL DISPERSANT EXPOSURE EXPERIMENT PROTOCOLS I

CRUDE OIL FUTURES OPTIONS MOCK TEST GUIDE 1 PARTICIPATE
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EXHIBIT A TO THE ISDA NORTH AMERICANUS CRUDE OIL

Preparing WAF

Crude Oil and Oil Dispersant Exposure Experiment Protocols


I. Preparing stock solutions of Water Accommodated Fraction of crude oil (WAF)

  1. Calculate the amount of WAF stock will be required for the experimental exposures (for each treatment container, prepare 50ml of WAF stock; exposures set up in small round plastic cups with lids).

  2. Vigorously stir oil with water in a 1:99 ratio using a magnetic stirrer for 2 hrs. The oil should be pipetted out of the bottle with a 1000ul pipet with the tip cut off. Since a significant amount of the oil will stick to the sides of the pipet, it is impossible to be 100% exact when measuring it out. Let stand for 24 hours in the preparation beaker to allow for the separation of oil and water phases (do not use sep funnel) (from Negri and Heyward 2000).

  3. For oil + dispersant mix, prepare solution in the same way using a 1:10 concentration of dispersant : oil (maximum recommended; see Negri and Heyward 2000). This solution will be noticeably cloudier than the oil only solution. It is best to use the WAF/CEWAF (Chemically Enhanced WAF- the oil with dispersant mixed in) as soon as possible, to minimize loss of volatiles (CROSERF).

  4. Dispersant only solutions should be prepared in the same way, replacing the volume of crude oil with sea water.

  5. Prepare ~50% extra stock solutions to allow for chemical sampling/assay. Keep in mind that the stock solutions will be diluted as well, so appropriate volumes need to be prepared.



II. Exposure experimental set – up

  1. A total of 18 wild anemones were harvested for both the WAF exposure and the CEWAF exposure.

  2. These were moved into six different treatment cups, with three anemones each in a control, 0.01%, 0.1%, 1%, 10%, and 100% WAF/CEWAF solutions. Since the WAF/CEWAF concentrations are a percent volume, the appropriate amount of sea water (e.g. 45 ml for the 10% treatment, 49.5 ml for the 1%) were pipetted into the containers at this time. The anemones were then let to attach for at least 24 hours by the aquaria.

  3. When dispensing the WAF/CEWAF, a glass or plastic pipet was used to extract the appropriate amount straight from the WAF/CEWAF preparation beaker. Because some crude oil stuck to the pipets (crude oil is very sticky), a kim wipe was used to minimize the transfer of raw, insoluble oil to the exposure containers. For the 100% WAF/CEWAF treatment, the exposure solution was simply poured from the preparation beaker into the exposure cup. This resulted in a small visible slick on the top of each 100% treatment.

  4. Treatment continued for 12 hours by the aquaria, after which the anemones were frozen for RNA extraction.


III. Some Useful Literature


Negri, A.P., Heyward, A.J.
Inhibition of fertilization and larval metamorphosis of the coral Acropora millepora (Ehrenberg, 1834) by petroleum products
(2000) Marine Pollution Bulletin, 41 (7-12), pp. 420-427.


-useful paper on exposing coral gametes and larvae to oil and corexit solutions. The protocol for making the WAF/CEWAF were taken from this paper, along with the exposure concentrations.



Mitchell, F.M., Holdway, D.A.
The acute and chronic toxicity of the dispersants Corexit 9527 and 9500, water accommodated fraction (WAF) of crude oil, and dispersant enhanced WAF (DEWAF) to Hydra viridissima (green hydra)
(2000) Water Research, 34 (1), pp. 343-348


-this paper presents a more straightforward toxicity testing design. They also include a different WAF preparation; everyone has their own special recipe.


Barron, M.G., Ka'aihue, L.
Critical evaluation of CROSERF test methods for oil dispersant toxicity testing under subarctic conditions
(2003) Marine Pollution Bulletin, 46 (9), pp. 1191-1199


-this paper contains modifications to the CROSERF WAF/CEWAF preparation guidelines that Negri and Heyward 2000 loosely follow. It is interesting only in an abstract sense, as much of what they discuss (and, indeed, the CROSERF guidelines themselves) are not possible in this lab, or at least too finicky to be worth the effort.


Note: The CROSERF guidelines – CROSERF stands for Chemical Response to Oil Spills: Ecological Effects Research, and is the body which has “standardized” toxicity testing of oil and dispersant compounds – can be found in the Oil Spill Dispersants: Efficacy and Effects book on page 200.


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FOR REFERENCE ONLY INE CRUDE OIL OPTIONS CONTRACT SPECIFICATIONS
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Tags: crude oil, of crude, protocols, exposure, experiment, crude, dispersant