Title 40

PART 300 APPENDIX C



Appendix C to Part 300 - Swirling Flask Dispersant Effectiveness Test, Revised Standard Dispersant Toxicity Test, and Bioremediation Agent Effectiveness Test

40:30.0.1.1.1.12.1.2.3 : Appendix C

Appendix C to Part 300 - Swirling Flask Dispersant Effectiveness Test, Revised Standard Dispersant Toxicity Test, and Bioremediation Agent Effectiveness Test Table of Contents 1.0 Introduction 2.0 Swirling Flask Dispersant Effectiveness Test 3.0 Revised Standard Dispersant Toxicity Test 4.0 Bioremediation Agent Effectiveness Test 5.0 Bioremediation Agent Toxicity Test 6.0 Summary Technical Product Test Data Format References List of Illustrations Figure Number 1 Swirling Flask Test Apparatus List of Tables Table Number 1 Major Ion Composition of “Instant Ocean” Synthetic Sea Salt 2 Test Oil Characteristics 3 Oil Standard Solutions: Concentrations in Final DCM Extractions 4 Synthetic Seawater [Toxicity Test] 5 Test Oil Characteristics: No. 2 Fuel Oil 6 Analytes Listed Under the Corresponding Internal Standard Used in Calculating RRFs 7 Primary Ions Monitored for Each Target Analyte During GC/MS Analysis 8 Analytes and Reference Compounds 9 Operating Conditions and Temperature Program of GC/MS 10 Two-Way ANOVA Table 11 Product Test Data, Total Aromatics 12 Summary Statistics for Product Test Data, Total Aromatics 13 Example Two-Way ANOVA Table 14 Pairwise Protected LSD Mean Separation 1.0 Introduction

1.1 Scope and Application. The methods described below apply to “dispersants, surface washing agents, surface collecting agents, bioremediation agents, and miscellaneous oil spill control agents” involving subpart J (Use of Dispersants and Other Chemicals) in 40 CFR part 300 (National Oil and Hazardous Substances Pollution Contingency Plan). They are revisions and additions to the EPA's Standard Dispersant Effectiveness and Toxicity Tests (1). The new Swirling Flask Dispersant Effectiveness Test is used only for testing dispersants. The Revised Standard Dispersant Toxicity Test is used for testing dispersants, as well as surface washing agents, surface collecting agents, and miscellaneous oil spill control agents. The bioremediation agent effectiveness test is used for testing bioremediation agents only.

1.2 Definitions. The definitions of dispersants, surface washing agents, surface collecting agents, bioremediation agents, and miscellaneous oil spill control agents are provided in 40 CFR 300.5.

2.0 Swirling Flask Dispersant Effectiveness Test

2.1 Summary of Method. This protocol was developed by Environment Canada to provide a relatively rapid and simple testing procedure for evaluating dispersant effectiveness (2). It uses a modified Erlenmeyer flask to which a side spout has been added for removing subsurface samples of water near the bottom of the flask without disturbing a surface oil layer. Seawater and a surface layer of oil are added to the flask. Turbulent mixing is provided by placing the flask on a standard shaker table at 150 rpm for 20 minutes to induce a swirling motion to the liquid contents. Following shaking, the flask is immediately removed from the shaker table and maintained in a stationary position for 10 minutes to allow the oil that will reform a slick to return to the water's surface. A sample of water for chemical analysis is then removed from the bottom of the flask through the side spout, extracted with methylene chloride (dichloromethane-DCM), and analyzed for oil content by UV-visible absorption spectrophotometry at wavelengths of 340, 370, and 400 nm (2).

2.2 Apparatus.

2.2.1 Modified Erlenmeyer Flask. Use 125-ml glass Erlenmeyer flasks that have been modified to include an attachment of a glass side spout that extends from the bottom of the flask upward to the neck region, as shown in Figure 1.

2.2.2 Shaker Table. Use a shaker table with speed control unit with variable speed (40-400 rpm) and an orbital diameter of approximately 0.75 inches (2 cm) to provide turbulence to solutions in test flasks.

2.2.3 Spectrophotometer. Use a UV-visible spectrophotometer capable of measuring absorbance at 340, 370, and 400 nm. A Hitachi Model U-2000 or equivalent is acceptable for this purpose.

2.2.4 Glassware. Glassware should consist of 5-, 10-, 25-, 100-, and 500-ml graduated cylinders; 125-ml separatory funnels with Teflon stopcocks; and 10-, 100-, and 1,000-ml volumetric flasks and micropipettes.

2.3 Reagents. 2.3.1 Synthetic seawater. The synthetic sea salt “Instant Ocean,” manufactured by Aquarium Systems of Mentor, OH, can be used for this purpose. The synthetic seawater solution is prepared by dissolving 34 g of the salt mixture in 1 liter of distilled water (i.e., a salinity of 34 ppt). Table 1 provides a list of the ion composition of the seasalt mixture.

Table 1 - Major Ion Composition of “Instant Ocean” Synthetic Sea Salt

Major Ion % Total Weight Ionic Concentration at 34 ppt salinity (mg/1)
Chloride (C1−) 47.470 18,740
Sodium (NA=) 26.280 10,454
Sulfate (SO4−) 6.602 2,631
Magnesium (Mg==) 3.230 1,256
Calcium (Ca==) 1.013 400
Potassium (K=) 1.015 401
Bicarbonate (HCO3−) 0.491 194
Boron (B) 0.015 6.0
Strontium (Sr==) 0.001 7.5
SOLIDS TOTAL 86.11% 34,089.50
Water 13.88
TOTAL 99.99%