NEW ENGLAND BIOASSAY, INC.

002049

TOXICITY REPORT

ENVIRONMENTAL SCIENCE CORPORATIONMiddletown, CT.

FOR

SOLVENTS RECOVERY OF NEW ENGLANDSouthington, CT.

January, 1990

Prepared By:

NEW ENGLAND BIOASSAY, INC.146 Hartford Road

Manchester, Connecticut 06040

NEW ENGLAND BIOASSAY, INC.

January 25, 1990

Mr. Thomas McGloin Environmental Science Corporation Great River Center 362 Industrial Park Road Middletown, Connecticut 06473

Re: Comparative Toxicity Testing Solvents Recovery of New England and Quinnipiac River

This letter report contains the results of bioassay tests performed on effluent from Solvents Recovery of New England, of Southington, CT, and Quinnipiac River water collected upstream of the facility on four consecutive days (15-18 January 1990). Definitive LC50 evaluations were performed on each of the eight samples with two species: the water flea, Daphnia pulex; and the fathead minnow, Pimephales promelas. The test procedures, described in Appendix A, were performed in accordance with Methods for Measuring the Acute Toxicitv of Effluents to Freshwater and Marine Organisms (Weber and Peltier, 1985, EPA/600/4-85/013) and specifications of the State of Connecticut, Department of Environmental Protection.

The test results, expressed as percent effluent, are summarized in the tables below. The raw survival and chemistry data from the tests on the river water and effluent samples are given in Appendices B and C, respectively.

Sample Species Day LC50 NOAEL

River D. pulex 1 >100 50 2 >100 100 3 >100 12.5 4 >100 100

P. promelas I >100 100 2 >100 100 3 >100 100 4 >100 100

146 HARTFORD ROAD / MANCHESTER, CONN. 06040 /TELEPHONE (203)643-9560

NEW ENGLAND BIOASSAY, INC.

Mr. Thomas McGloin January 25, 1990 Page 2

Sample Species Day LC50 NOAEL

EffluenEffluentt DD.. pulepulexx 11 2 3 4

>10>1000 92.4

>100 >100

5500 50 100 25

P. promelas 1 2 3 4

>100 >100 >100 >100

100 100 100 100

The Quinnipiac River samples were clear and colorless. They were slightly supersaturated with dissolved oxygen, and their pH values were near neutrality. All four samples were nontoxic to the fathead minnow, but the samples from days 1 and 3 were toxic to the water flea.

The effluent samples were yellow and turbid, and those from the first two days had an oily odor. The day 1 and day 2 samples were supersaturated with dissolved oxygen, whereas the others were below saturation. The day 2 sample was considerably more alkaline than the others (pH 8.1). All four samples were nontoxic to the fathead minnow, but those from days 1, 2 and 4 were toxic to the water flea.

I appreciate your using New England Bioassay for your aquatic toxicity testing, and look forward to working with you again. Please call if you have questions or need additional information.

Carol E. Bower Director

CEBrklb 190-07 CTT:SRONE01

NEW ENGLAND BIOASSAY INC.

APPENDIX A

Acute Freshwater Methods (CT)

METHODS FOR ACUTE TOXICITY TESTING WITH FRESHWATER ORGANISMS

(Connecticut)

Unless otherwise noted, all test protocols follow those of the Connecticut Department of Environmental Protection (DEP) and the U.S. Environmental Protection Agency (EPA) as specified in Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms. 3rd ed. (1985, W.H. Peltier and C.I. Weber, eds., EPA/600/4-85/013).

Effluent Collection and Handling

Grab or composite effluent samples are collected by personnel at the client's facility, refrigerated until pick-up, and then transported by NEB personnel to the testing laboratory. Subsamples are submitted to an approved analytical laboratory when appropriate.

Toxicity Testing

TEST SPECIES:

The test organisms are the water fleas, Daphnia pulex and Daphnia magna. and the fathead minnow, Pimephales promelas. All species are reared at New England Bioassay from stocks originally obtained from the Connecticut DEP or USEPA. The ages of the Daphnia and fish are 0-24 hours and 25-35 days, respectively, at the start of testing. Fish from a single day's hatch are used whenever possible.

DILUTION WATER:

Artificial freshwater (deionized, activated-carbon-filtered water reconstituted with sodium bicarbonate, potassium chloride, calcium sulfate, and magnesium sulfate) is prepared in the New England Bioassay laboratory. The pH is 7.0 to 7.6, and the hardness is approximately 50 mg/L as calcium carbonate, unless otherwise specified for tests conducted with D. pulex and P. promelas. High-hardness water (160 mg/L) is used in bioassays in which D. magna is the test organism.

WATER QUALITY DETERMINATION:

Sample temperature, pH, conductivity, hardness and concentration of dissolved oxygen are measured initially and daily throughout testing, in accourdance with standard procedures (American Public Health Assoc. et al., 1985, Standard Methods for the Examination of Water and Wastewater. 16th edition). Temperature and conductivity are measured with YSI Model 33 S-C-T meter;

Acute Freshwater Testing (CT) Page 2

dissolved oxygen is measured with YSI Model 51B meter; pH is measured with Beckman Model 12 pH/ISI meter; and hardness is determined by the EDTA titrimetric method.

GENERAL TEST CONDITIONS AND PROCEDURES:

The bioassays are static renewal or non-renewal tests. In non-renewal tests, the test organisms are exposed to the same solution for the entire test period. In renewal tests, the test solutions are replaced with fresh solutions of the same concentrations at a specified interval or intervals during testing.

All tests are performed at ambient temperature (20-22°) with a photoperiod of 16 hours of light and 8 hours of darkness. Refrigerated solutions are brought to ambient temperature before testing. Test durations are 48 hours CDaphnia species) or 96 hours (fish), unless noted otherwise. When the initial concentration of dissolved oxygen in the undiluted sample is less than 40% of the saturation value, samples are aerated before the organisms are introduced and subsequently during testing; otherwise no supplemental aeration is provided during testing. Ten organisms are used in each exposure chamber in each test. The organisms are not fed during testing.

The measured effect in each test is death or immobility, evidenced by an organism's failure to respond to gentle prodding. At 24-hour intervals throughout testing, survival data are collected and recorded, and dead organisms are removed from the test chambers.

REFERENCE TOXICANT:

Copper nitrate, obtained from the Water Compliance Unit (WCU) of the Connecticut DEP, is used as a standard to authenticate the sensitivity of the laboratory stocks of Daphnia and fish. Definitive tests with the reference toxicant are performed weekly, concurrent with effluent bioassays, according to standard protocol (see below).

Specific Toxicity Tests

DEFINITIVE TEST:

In a definitive test, organisms are exposed in duplicate chambers to five, two-fold dilutions of effluent (e.g. 6.25, 12.5, 25, 50, and 100%). Control organisms are exposed in duplicate chambers to dilution water (0% effluent). A

Acute Freshwater Testing (CT) Page 3

definitive test is used to determine the median lethal concentration (LC50) of an effluent (see "Statistical Analysis of Data" below).

RANGE-FINDING TEST:

A range-finding test is an abbreviated test for preliminary determination of effluent toxicity. In this test, organisms are subjected in single exposure to four or five widely-spaced effluent dilutions (e.g. 1, 10, 25, 50 and 100%) and compared with control organisms in dilution water. The duration of range-finding tests can vary from 24 to 96 hours, depending on the species used and the reason for performing the test.

SCREENING ("PASS/FAIL") TEST:

The Connecticut DEP stipulates the use of screening ("pass/fail") tests for industrial cooling waters and municipal discharges. In a screening test, a single effluent concentration, usually 100%, is tested in five replicates against three replicate controls (dilution water). Mean survival of 90% or greater in the test solution constitutes a "pass" score, provided that 90% or more of the control organisms in each test chamber also survive. If an effluent fails a screening test, dilutions generally are prepared and a definitive test is performed.

pH ADJUSTMENT TEST:

In pH adjustment tests, undiluted wastewater is adjusted to two pH values, 6.0 and 8.5, and then screening tests are performed. These tests are designed to determine whether ammonia is the primary toxicant in the effluent. The principle underlying this test is that in water, ammonia exists in two forms, ionized and free (un-ionized), the relative proportions of which are determined mainly by pH. The higher the pH, the greater the proportion of free ammonia; each increase of one pH unit increases free ammonia by a factor of 10. Free ammonia, a poison, is the only form that can readily enter into living cells and tissues. It is thus expected that any given concentration of total ammonia (ionized plus free) will be more toxic at a higher pH value, where the free form predominates.

Acute Freshwater Testing (CT) Page 4

Statistical Analysis of Data

All computer programs for statistical analysis of data were obtained from the Connecticut DEP/WCU, and are identical to those used by the WCU. Mortality data are analyzed statistically by four different methods to determine the "median lethal concentration" (LC50) and the "no observed acute effect level" (NOAEL) of the effluent. The LC50 is the concentration that is lethal to 50% of the organisms within the test period. The NOAEL is the highest concentration at which 90% or greater of the organisms survive. Either or both of these values is used for calculating how much effluent can be discharged safely into a given stream.

The computational methods used to calculate the LC50 values are: i) binomial distribution; ii) moving average-angle; iii) probit; and iv) trimmed Spearman-Karber. The value with the "best statistical fit", i.e. that which best matches the raw data and has the most narrow 95% confidence interval (the range of values within which the true LC50 value could occur 95% of the time), is then selected.

FORMS:METHCT-FW REV 12/89

NEW ENGLAND BlOASSAY, INC

APPENDIX B

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APPENDIX C

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