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Volume III, Appendices EM - National Marine Fisheries Service ...

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Appendix 7. Petroleum Hydrocarbon Tissue Sampling Protocol<br />

Supplies for sampling<br />

All instruments used in handling (e.g. scalpels and forceps, cutting boards) or storing (e.g. jars, foil,<br />

sheets) samples must be made of a non-contaminating material consisting of stainless steel, glass,<br />

Teflon, or aluminum.<br />

- Solvent-rinsed glass containers with Teflon-lined lids for tissues<br />

- Solvent-rinsed Teflon sheets for tissues<br />

- Aluminum foil (if Teflon sheets are not available) sample to the dull side<br />

- Sterile syringes and needles<br />

- Amber glass vials or glass vials covered with foil with Teflon lids (for bile, urine)<br />

- Teflon screw top vials (for blood storage and urine)<br />

- Stainless steel scalpels, knifes, forceps<br />

- Isopropyl alcohol (99.9% pesticide free IPA) to rinse instrument<br />

- Wooden tongue depressors (can be used to handle tissues if necessary)<br />

- Whirl-pak bags or Zip-lock freezer bags<br />

- 10% buffered formalin and appropriate containers for histopathology samples<br />

- Permanent marker or pen<br />

- Evidence/Custody tape and labels<br />

- Sample Log/Chain of Custody forms<br />

Sampling Protocol<br />

Tissues to collect for petroleum hydrocarbon analysis in order of preference:<br />

a. bile<br />

b. urine<br />

c. whole blood<br />

d. stomach and intestinal contents<br />

e. blubber/fat<br />

f. liver<br />

g. kidney<br />

h. lung<br />

i. intestine<br />

j. brain<br />

k. muscle<br />

i. Samples taken for analysis should only be collected from alive or freshly dead<br />

animals. If a necropsy cannot be performed within 24 hrs after death, the carcass<br />

should be frozen for later examination.<br />

ii. Recommended minimum sample size is 10-20 g of tissues (approx. 1-2<br />

tablespoons) and 5 ml for fluids (blood, urine, bile, feces, stomach contents).<br />

However, analysis can be performed on as little as 100 L of bile; therefore collect<br />

whatever amount is present.

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