ML17223B308
| ML17223B308 | |
| Person / Time | |
|---|---|
| Site: | Saint Lucie  |
| Issue date: | 06/10/1991 |
| From: | AQUA SURVEY, INC. |
| To: | |
| Shared Package | |
| ML17223B307 | List: |
| References | |
| 91-089H, 91-89H, BR91-1513H, NUDOCS 9111140225 | |
| Download: ML17223B308 (40) | |
Text
, ATTACHMENT3, RECUR SURVEY, INC.
FINAL REFOPT THE ACUTE TOXICITY OE'ET?i CL 788-8700 FAX (908> 788-9165
RECUR SURVEY, INC.
STATEMENT OF NO CONFIDENTIALITY CLAIMS No claim of confidentiality is made for any information contained in this study on the basis of its falling within the scope of FIFRA Section 10 (d)
(1)
(A),
(B) or (C).
BETZ LABORATORIES, INC.
4636 Somerton Road
- Trevose, PA 19047 r
r Company Agent atic Toxicolo ist Laborator ana er Title Date Signature ASI Project Number:
91-089H 2 of 11 CLll 499 Point Breeze Roost
~ Flernington. New Jersey 08822
~ Telephone(908) 788-8700 FAX (908> 788-9155
7
~
7 FIQUR SURVEY INC THE ACUTE TOXICITY BIOASSAY OF CLAM-TROL CT-1 This study meets the requirements for 40 CFR Part
- 160, however dilution water chemical/physical characteristics were performed by Nytest Environmental, Inc. and might or might not meet these requirements.
Submitter:
Signature:
Betz laboratories, Inc.
4636 Somerton Road
- Trevose, PA 19047 Date Sponsor:
Signature:
Betz Laboratories, Inc.
4636 Somerton Road
- Trevose, PA 19047 Date Study Director:
ork Terrell D te 3 of 11 CL/I rt(rr) Point Breeze Road
~ Flemington, New Jersey 08822
~ Tetephone (908) 788-8700 FAX (O08) 788.O)85
R(j)UFI SURVCY, INC.
Summary A sample of Betz Clam-Trol, CT-l, received from Betz Laboratories, Inc., Somerton
- Road, Trevose, PA 19047 on March 21,
- 1991, was tested for potential acute toxicity to E'" "'"'
""'"'onducted under static conditions with daily renewal of the test solutions at nominal concentrations of 0.625, 1.25, 2.5, 5.0 and 10.0 ppm.. Concentration validation made before renewals at TO and T73 revealed a mean concentration of 0.738, 1.411, 2.834, 5.968 and 11.980 ppm while concentration validation made after renewals at T23 and T96 revealed a mean concentration of 0.490, 1.043, 2.545, 5.091 and 11.402 ppm.
After 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> mortality ranged from 254 at the 0.625 ppm concentration to 1004 at the 5.0 and 10.0 ppm concentrations.
The nominal 96 hour0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> LC50 was 1.24 ppm calculated by the Probit Method.
4 of 11 CLll 499 Point Breeze Road
~ Flemington. New Jersey 08822
~ Telephone (908) 788 8700 FAX (908) 788 9i65
RECUR SURVCY, INC.
I.
Ob)ective The ob)ective of this study was to determine the acute toxic effects of Betz CT-1 in an aquatic environment and to establish the 96 hour0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> median lethal concentration (LC50) to g. ~b ogius under static daily renewal conditions.
II.
Test material Source:
Name:
Density:
4 Solubility in Water:
Betz Laboratories, Inc.
Somerton Road
- Trevose, PA 19047 Clam-Trol~,
(CT-1) 100 Date Received:
March 21, 1991 Amount Received:
1 pint III. Materials and Methods A.
Method The method employed was a modification of the method outlined in Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms EPA/600/4-85/013 March 1985.
(See Appendix A for the Protocol and Appendix B
for the SOP).
B.
Test Organisms
~seciee The test species, representing a typical temperate p
4
- rrwwwww e "
5 of ll CLll 4r)P Pornt 8reeze Road
~ Flernington, New Jerzey 08822
~ Telephone (r)08) 788-8700 FAX(F08) 788-9165
~,
RCj)UR SURVCY, INC.
e e
sic o
t o Animals used for this test were mixed adult amphipods.
Animals appeared to be healthy and in good physical condition.
So c
cc a
o All animals utilized in this test were from Northwestern Aquatic Sciences, Inc. of Newport, Oregon.
The animals were acclimated to dilution water and test temperature for six days.
C ~
Test System ce of o Wate Water utilized for this test was filtered (0.45 microns) natural seawater obtained from Manasquan River Inlet, Manasquan, New Jersey.
(See Appendix D-1 for water characteristics.)
e eratur The test temperature was 15 +/- 34 C
The test vessels were 1 liter, 10 x 14 cm glass beakers containing 500 ml of test solution at a
depth of 7 cm.
o o er o A 24-hour light/0-hour dark photoperiod with utilized.
~oc~
Each test chamber contained ten organism in 500 ml of test solution, resulting in a loading factor of one organism per 50 mls of test solution.
6 of 11 CUI 499 Point Breeze Road
~ Flemington. New Jersey 08822
~ Telephone (908) 788-8700 FAX (908) 788-9168
RECUR SURVEY, INC.
D.
Test Design A range finding test was conducted separately.
Four concentrations of the test material were evaluated using 5 amphipods per treatment.
The definitive test was conducted using 10 organisms in each of 2 replicates at each of five treatment levels.
~ao t ol A control treatment was run concurrently with the test substance treatments using the same dilution water and the same number of organisms as per the test concentrations.
es
'tiat o Testing was conducted under static conditions with daily renewal of the test solutions.
An initial test material stock solution of 1,000 ppm was prepared with ASI DI water.
Test solutions were prepared from the stock solution and then dispensed into the test chambers.
The test was initiated when, within 30 minutes of test solution preparation, the test organisms were transferred to the test chambers.
The range-finding test spanned 72 hours8.333333e-4 days <br />0.02 hours <br />1.190476e-4 weeks <br />2.7396e-5 months <br /> and was conducted using 5 organisms at each of four treatment levels:
1.0, 10.0, 100 and 1000 ppm.
Test results indicated a toxic response between 1.0 and 10.0 ppm.
Exposure at the lower concentration level resulted in 204 mortality after 72 hours8.333333e-4 days <br />0.02 hours <br />1.190476e-4 weeks <br />2.7396e-5 months <br />, while exposure at the higher level resulted in 1004 mortality after 72 hours8.333333e-4 days <br />0.02 hours <br />1.190476e-4 weeks <br />2.7396e-5 months <br />.
Results from this test were used to determine the treatment levels for the definitive test.
The concentrations used were 0.625, 1.25, 2.5, 5.0 and 10.0 ppm.
The definitive test was for 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> (4
days) duration and was conducted using two (2) re-replicates of ten organisms at each of five treatment levels.
7 of ll CVI 499 Point Breeze Road
~ Flemington, New Jersey 08822
~ Terepnone (908) 788-8700 FAX <908> 788-9168
FlQUFl SURVEY, INC.
Test vessels were examined at each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> interval from test initiation through test termination.
Survival/mortality determinations were recorded for each chamber.
Dead animals were removed and discarded.
C e ica A
ses Concentrations were validated at TO, T23, T73 and T96 using Clam-Trol~,
CT-1 methyl orange method (supplied by Betz Lab) in the range of 0.2 5.0 mg/l.
A Bausch 6 Lomb Spectronic 21 was used for the absorbance readings.
(see appendix, page C-l.)
s'c 1
C e ic a ete The temperature, pH, dissolved oxygen (DO) concentration and salinity were measured and recorded for each treatment, level at the beginning of the test and at each 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> test interval thereafter.
IV.
Results The definitive test resulted in mortality ranging from 254 at the 0.625 ppm concentration to 100%
at the 5 and 10 ppm concentrations (See Table 1).
The test solution temperature was maintained at 15.0 C.
(See Table II).
Dissolved oxygen was maintained between 6.8 and 8.2 mg/l.
(See Table III).
The pH ranges during the test were between 7.7 and 7.8 (See Table IV).
~
The mean concentration validation before each renewal period at TO and T73, and at the end of each renewal period at T23 and T96 can be seen in Table V.
V.
Source of Documentation All original data documentation is being maintained.at Aqua Survey, Inc.,
499 Point Breeze
- Road, Flemington, NJ 08822, in Betz Laboratories Notebook 002, pages 81-92.
8 of 11 499 Point Breeze Road
~ Flemington, New Jersey 08822 i Telephone (908) 788-8700 FAX (908) 788-9165
RECUR SURVEY, INC.
Table I:
o t D a Exposure Time
~Co t~o g 6~
(hrs. )
A B
A B
A B
A B
A B
~0 A
B 0
0 0
0 0
0 0
0 0
0 0
0 0
0 0
0 0
10 0
0 0
0 90 90 0
0 8
70 00 00 96 10 0
30 0
50 70 0
0 Table II e
e tu ees Exposure Time
~Co t ol (hrs.)
A B
A B
A B
~50 A
B A
B
~0 A
B 0
0 0
15 0 15.0 5
0 0
5.0 5
0 5
0 0
5 0
15 0 15.0 15 0
5.0 15 0
15 0 15.0 5.0 15 0
5 0
5 0
5 0 15.0 15.0 5
5 0
0 5
5 0
.0 72 96 Table III 15 5
0 5
0 0
5 0
15 0
5 5
0 5
0 15 0
5 0
5 5
Dissol ed 0 e
m 1
0 0
Exposure Time
~co t~o
~06 5
(hrs. )
A B
A B
A B
A B
A B
~0 A
B 0
0 8
0 8
0 8.0 8
0 7
8 7
8 7
7 7
8 7
8 7
7 8
7 8
7 7
0 0
0 0
0 0
0 8
8 8
8 8
8.
72 8
1 8
8 8
96 7
3 7.3 7
0 7
0 7
0 6
9 6
8 6
7 Table IV Exposure Time
~Cont o
(hrs.)
A B
0.625 A
5 A
B
~5 A
B
~0 A
B
~00 A
B 0
0 8
7 8
8 7
8 7
7 7
7 8
8 7
7 8
8 7
8 7
7 7
7 7
8 8
7 7
8 7
7 8
7 8
7 8
8 7.8 7
7 8
72 7
8 7
8 7
8 8
7 8
7.8 7
8 7
8 8
7 8
6 7
7 7
7 7
7 7
9 of ll CVl 499 Point Breeze Road
~ Flernington, New Jersey 08822
~ Teiephone (908) 788-8700 FAX (908) 788-9165
RECUR SURVCV, INC.
Table V
a 'o Conc.
ppm 0.625 1.25 2.5 5 '
10.0 0.749 1.305 2.695 5.901 11.980 0.726
- 1. 516 2.973 6 '34 Mean 0.738 1.411 2.834 5.968 11.980 o ce tration Vali atio fte e
ew Conc.
ppm Mean 0.625 1.25 2.5 5.0 10.0 0.498 1.137 2.528 5.056 11.402 0.482 0.949 2.561 5.126 0 ~ 490 1'43 2.545 5'91 11.402 10 of 11 CVI 4ort Point Breeze Road
~ Flemington, New Jersey 08822
~ Telephone (r708) 788 8700 FAx (o08> 788 oio8
"ONCENTRATZON 10.0000 5.0000 2.5000 1.2500 0.6250 NUMBER EXPOSED 20 20 20 20 20 NUMBER AFFECTED 20 20 15 9
5 PERCENT AFFECTED 100.0000000 100.0000000 75.0000000 45.0000000 25.0000000 BINOMIAL PROBABILITY
(~)
0.0000954 0.0000954 2.0694730 41.1901500 2.0694730 METHOD SPAN RESULTS EC50 95 PERCENT CONFIDENCE LIMITS BINOMIAL 1.397710 0.625000
2.500000
'DROVING AVERAGE PROBZT 0.109754 0.130604 1.295583 1.238264 0.927734
1.675829 0.903236
1.602543 INTERPRETATION OF STATISTICS The order of preference of results is:
BEST:
Probit Method BETTER: Moving Average GOOD:
Binomial GOOD:
Graphic Interpolation (Graph drawn by hand) assuming all are statistically valid.
To Determine if a method has given valid results:
GRAPHICAL:
Always valid BINOMIAL:
The program will tell you if this is not valid.
MOVING AVERAGE: The number of spans needs to be no less than one lower than the number of concentrations tested (excluding control).
PROBIT:
The program will tell you if this is not valid Page 11 of 11
A P
P N
D I
RECUR SURVe, lac.
A8Z MOc 9100 '7 Map 1i < 1991 PROTOCOL ACUTE TOXICITY BIOA88AY POR ~o~s SPONSOR:
0 6
6 om o
o vo e 0
7 STUDY NUMBER:
-0 9
TESTING FACILITY:
Aqua Survey, Inc.
(AS I) 499 Point Breeze Road Flemington, New Jersey 08822 United States of America PROPOSED START DATE:
a 20 9
PROPOSED COMPLETION DATE:
a 27 1991 STUDY DIRECTOR QA OFFICER:
SPONSOR APPROVAL:
Signature
)
Sinure Signature Dat'e f>>;
Date Date 1 of 6 499 Potnt 3reeze Hoed t=!en.in@ion,."ew
.'-..e'l,::"'""
'e.-.","e 9Q3)7ac.87CO V X:,9C""1733-~iQ=
?Q.
Box 3"059
~ Ba;on Rouge. iou)5lono:C3r
<!eon ne (3QQ) I' 4534 FAX ( 04j 92'93'
RECUR SURVqy, igC EUREOSE To determine the acute toxicity of the test material in the aquatic dIII I
I IY I
conditions for 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br />.
TEST ARTICLE The test article is ~ which was received from et bs c.
on 1YIIILtl' (description).
The sample is labeled with an ASI number ~~0 Any test article remaining after the test is completed will be disposed of according to instructions from the sponsor or returned to the sponsor.
TEST ARTICLE CHARACTERISATION:
Data concerning stability, uniformity, composition and additional chemical/physical characteristics of the test article will be provided by the sponsor on the Test Substance Form (supplied by ASI).
Chemical analyses will be the responsibility of the sponsor unless prior arrangements have been made for ASI to assume that responsibility.
CONTROL ARTICLE None required.
TEST SYSTEM "I
estuarine/marine amphipod.
JUSTIPICATION "I
I are a characteristic system in which the response to toxicological agents can be evaluated.
DILUTION RLTER Whenever possible, natural sea water will be used.
If use of natural seawater is not possible, an alternative dilution water, uncontaminated and of constant quality, will be used.
SOLVENTS Whenever possible, the toxicant will be introduced into the test solution without the use of solvents other than water. If alternative solvents are necessary, they will be used sparingly, not to exceed 0.5 ml/l in a solution.
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TEST MATERIALS The technical grade, end-use product or both will be used in this study.
The brand name or chemical names and a lot number are used to define a
test material.
Mixtures are defined by ingredient and percent of each ingredient, where possible.
TEST ORGANISMS' l
p d
organisms will be as uniform as possible in age and size.
The age of the organisms will be ascertained from the supplier along with any disease treatment schedule.
EXPOSURE CHAMBER One (1) liter all glass beaker or other appropriate size glass chambers will be used.
RANGE-FINDING TEST:
If the toxicity of the test substance is not already
- known, a range finding test will be performed to determine the range of concentrations to be used in the definitive test.
DEFINITIVE TESTi The purpose of the definitive test is to determine the concentration-response curves and the 48 and 96 hour0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> LC50 values.
A minimum of 20 organisms per concentration vill be exposed to five or more concentrations of the chemical.
Each designated treatment group vill be exposed to a concentration of toxicant that is at least 504 of the next highest concentration.
Test organisms will be divided into two chambers per treatment, group (10 organisms each).
All organisms will be randomly assigned to test vessels.
TEST PROCEDURES In preparation for the test, the chambers are filled with appropriate volumes of dilution water.
The test substance is introduced into each chamber.
The test is started by randomly introducing organisms acclimated in accordance vith the test design into the test chambers within 30 minutes after the addition of the test substance.
Organisms in the test chambers are observed periodically during the test, the dead animals removed and the findings recorded.
Dissolved oxygen concentration, pH, temperature and other water quality characteristics are measured at specified-intervals in test chambers.
The test. solution will be renewed daily.
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ACCLZMATXONc Organisms vill be acclimated to the dilution vater as long as possible.
The organisms will receive a standard diet daily until just prior to testing, at which time feeding will be discontinued.
Organisms will not be fed during test.
Test organisms will be observed prior to testing for signs of disease,
- stress, physical damage and mortality.
- Injured, dead and abnormal individuals will be discarded.
If organisms show any of these signs, remedial action will be taken during the acclimatization period and not during the test phase.
Organisms that have been treated for disease are acceptable for testing provided they do not exhibit signs of disease during the 48 hours5.555556e-4 days <br />0.0133 hours <br />7.936508e-5 weeks <br />1.8264e-5 months <br /> of pretest.
Organisms will not be used if they appear to be diseased or stressed or if more than 34 die during the 48 hours5.555556e-4 days <br />0.0133 hours <br />7.936508e-5 weeks <br />1.8264e-5 months <br /> immediately prior to testing.
S ~
TEMPERATURE:
LaaEI1!a
+/
measured daily in at least one test vessel of each test concentration as long as organisms are surviving at that level.
CONCENTRATION MEASUREMENT:
Analytical measurements of the test substance will be made for each concentration at To, T>, Tz, and T9< of the test by a method valida'ted before beginning the test by appropriate laboratory practices.
pie The pH will be measured in at least one test vessel at the beginning of the test and daily thereafter at each toxicant concentration as long as animals are surviving. at that level.
SALINITY:
The salinity will be measured in at least one test vessel at the beginning of the test and daily thereafter at each toxicant concentration as long as animals are surviving at that level.
DZSSOLVED OXYGEN Dissolved oxygen (DO) will be measured at the beginning of the test and daily thereafter.
Measurements will be taken from each concentration as long as animals are surviving at that concentration.
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~Nq PHOTOPERZODs A 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> light and 0 hour0 days <br />0 hours <br />0 weeks <br />0 months <br /> dark photoperiod will be employed.
LOADINQ:
The size of the test vessels will be such that the loading factor is no greater than 1 organism/10 ml of test solution.
OBSERVATIONS:
The organisms will be observed at. 24, 48, 72, and 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> with particular attention being given to general behavior and mortality.
A study is not acceptable if more than 104 of the control organisms die during a test.
REPORT:
The following are included in the final report:
Test Material - If technical grade product is tested, the
- source, batch and exact purity are reported.
When end-use product is tested, the exact percent of active ingredient and the type of formulation (i.e. granular, wettable powder) are reported.
2 ~
Dilution Rater Dilution wate source, its chemical/physical characteristics, and pretreatment are reported.
3 ~
Exposure Chambers The volume, diluent depth, and container construction material are reported.
4 ~
Test Organisms Source, quarantine, holding and acclimatization, inclusive of feeding schedules and disease treatment procedures, are reported.
The age and scientific names are reported.
Range Pinding Test Procedures and results are reported for the range finding study.
Included at a minimum are sample size, concentrations tested and mortality data.
6 ~
Definitive Test - Procedures used to prepare toxicant stock solutions are described.
The method of dosing is reported.
The criteria for determining effects are defined.
-Raw data or percentage of deaths/effects at each treatment level along with the number of organisms exposed at each level is reported.
Toxic
- symptoms, both physical and behavioral, observed during the test are report:ed.
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Calculato4 LCSO Value - LC50 values (48 and 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br /> if possible) with the corresponding 954 confidence intervals and slopes of the concentration response lines and method of calculation will be reported.
8.
Temperature/DO/pH/salinity - Temperature, dissolved
- oxygen, pH and salinity measurements are reported along with the range and mean of temperature.
9.
Chemical Analyses - When chemical analyses are made to validate toxicant concentrations, the analytical method is reported as well as the results.
Residue observed in exposure chambexs is described in the report.
10'esting Protocols This protocol is referenced in the report.
11.
Deviations - Deviations fxom protocol or standard operating procedures are reported.
2.2.
Certification - The Study Directox and the QA Manager certify the final report.
RECORDS'll raw data, data manipulations, draft reports and final reports will be retained by Aqua Survey, Inc. for a period of five years from the date of completion of testing.
Records will be surrendered to the sponsor upon their written request.
QUALXTY ASSURANCE:
This study will be conducted in accordance with the EPA Good Laboratory Practice Regulations (Federal Register, Volume 48, November 29, 1983) and Methods.for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms EPA/600/4-85/013 March 1985,.
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ASZ HO 625 June 14, 1991 STAHDARD OPERATZNQ PROCEDURE Acute Toxicity Bioassay for Amphipods (Daily renewal)
Marino and Estuarine The objective of this is to determine the acute toxicity of the test article in the aquatic environment to Marine and Estuarine Amphipods under static daily renewal conditions for 96 hours0.00111 days <br />0.0267 hours <br />1.587302e-4 weeks <br />3.6528e-5 months <br />.
1.
Sufficient volume of filtered (0.45 microns)
Manasquan sea water properly prepared and stored; 2.
Approximately 200 amphipods as uniform as possible in age and size; 3 ~
20 1000 ml all glass beakers or other appropriate size glass chambers; Appropriate size chiller and chiller bath or other chiller unit; 5.
Appropriate volume of DZ water; 6.
One stainless steel 5-inch diameter or other appropriate size sieve; 7.
Several'isposable clipped (5-mm bore) pipettes; 8.
Data collection forms; 9.
Temp mentor; 10'lass thermometers (graduated 0.5'C);
11.
DO meter; 12.
pH meter; 13.
Salinity refractometer; 14.
6 4-liter or other appropriate size mixing chambers; 1 of 3
15.
One(1) and 10 ml graduated glass pipettes; 16.
2 100-ml or other appropriate size graduated cylinders; 17.
One 1000-ml or other appropriate size volumetric flask; 18.
40 fathoms sea salt; and 19.
Analytical balance.
u e 1.
Test Article Preparation a.
Stock solution Fill volumetric flask approximately half full with DZ water.
Measure or weigh out appropriate amount of test article using glass pipettes or analytical balance.
Place test article in volumetric flask and bring up to volume with DZ water.
Place stir bar in flask and place on star plate for approximately 5 minutes or until sample is thoroughly mixed.
The stock solution is usually 1,000 or 10,000 ppm.
b.
Test solution Dilution water (filtered Manasquan sea water) should be adjusted to the appropriate salinity with DZ water and 40 fathoms sea salt if needed.
The temperature should be adjusted by refrigeration or water bath.
Appropriate volumes of dilution are then measured using graduated cylinders and placed in mixing chambers where appropriate volumes of stock solution are added and placed on a stir plate and allowed to thoroughly mix.
Appropriate volumes of the test solutions are then poured into the test chambers (labelled according to concentration and replicate and pre-rinsed with the appropriate test solution) and placed into the water bath.
2 ~
Range-Finding Test Xf the toxicity of the test article is not known, a range-finding test will be performed to determine the range of concentrations to be used in the definitive test.
Test concentrations used for the range finding test are usually
.1, 1,
10, 100 and 1,000 ppm, using one replicate of 5 organisms each.
2 of 3
3.
Definitive Test All organisms are randomly assigned to test vessels.
Carefully pour sand with organisms (acclimated according to protocol) on to a stainless steel sieve and placed in a vessel of dilution water.
Allow the sand to filter through the sieve, leaving the amphipods in the sieve suspended in the water.
Organisms are then placed two in each test vessel using a clipped pipette until each vessel contains 10 organisms.
4.
Daily Reading All readings are taken in accordance with the protocol from the mixing chambers prior to being placed in the test chambers with the exception of the final reading which is taken from the test chambers.
6.
Feeding None.
Daily renewals 4
Test chambers are carefully cleared of all detritus and dead organisms using a disposable pipette.
Each chamber is then gently decarted into a clean chamber until approximately 100 ml of test solution remains in the chamber, taking care not to pour off any of the test organisms.
Survival/mortality counts and behavioral and morphological observations are made at. this time or in accordance with protocols.
Appropriate amounts of new test solutions are carefully poured into the exposure chambers so that the solution runs down the inner chamber walls.
7 ~
Reporting All test data will be reported in accordance with requirements in the protocol.
3 of 3
CLAM-TROL CT-1 METHYLORANGE METHOD APPARATUS REQUIRED Code Cylinder, graduated, 100 mL 121 Cylinder, graduated, 250 mL Flask, volumetric, 1 L, glass (4 required) 935 Plpet, glass, graduated, 1 mL 140 Plpet, glass, volumetric, 1 mL 888 Pipet, glass, volumetric, 3 mL OO Pipet, glass, volumetric, 5 mL 124 Plpet, glass, volumetric, 10 mL 123 Pipet, glass, volumetric, 15 mL 881 Pipet, glass, volumetric, 20 mL Pipet, glass, volumetric, 25 mL 117 Plpet, glass, volumetric, 30 mL The general apparatus required for the test ls deter-mined by the specific test procedure used.
Apparatus not available through Betz Lab Supply should be obtained through a local supplier.
917 CHEMICALS REQUIRED 1, 2-Dichloroethane (reagent) grade or equivalent)
Code 1668 Beaker, glass, 50 mL (2 required)
Code Cylinder, graduated, 25 mL 2822 Funnel Rack, separatory 938 Funnel, separatory, with a Teflon stopcock, 250 mL ~~
(2 required)
Glass Rod 114 Optical Cell, (2 required)
Spectrophotometer GENERAL APPARATUS ~
CT-1 Buffer Reagent Methanol (reagent grade or equivalent)
Drying Reagent, with a plastic dipper
SUMMARY
OF METHOD 1591 1271 GENERAL PROCEDURE Use a well-ventilated or hooded area to run the test.
Always use a safety bulb when plpettlng Ilqulds.
1,2-Dlchloroethane (also known as Ethylene Dichlo-ride) is a priority pollutant and a specifically-listed RCRA~egulatedmaterial subject to specific disposal re-strictions and/or prohibitions. For this reason, all used 1,2-dichloroethane should be segregated from other waste streams. Dispose ofwaste 1,2-dlchloroethane in an approved manner (e.g., labpacklng or incineration).
In this procedure the dye inthe CT-1 Buffer Reagent com-plexes with the active ingredients in Clam-Trol CT-1. This complex is extracted into 1, 2-dlchloroethane. The or-ganic layer containing the complex is separated from the aqueous layer and driedwith a drying reagent containing anhydrous sodium sulfate. The color intensity ofthe 1, 2 dichloroethane layer is then measured in a spectropho-tometer at 415 nm.
This method must be customized to each specific appli-cation. Varythe volumes of sample, CT-1 BufferReagent, and
- 1. 2 - dlchloroethane according to the test range (see Table 1). Ifa higher absorbance is needed, increase the volume of sample or decrease the volume of 1, 2-olcnioroethane. When increasing the sample volume it may be necessary to increase the volume of CT-1 Buffer Reagent used. For samples <150 mLuse 10 mLof CT-1 Buffer Reagent; for samples between 150 and 300 mL use 15 mL of CT-1 Buffer Reagent.
Make sure that enough 1, 2 - dlchloroethane is used to leave a small plug of solvent in the separatory funnel when the bottom layer of solvent is removed and to fill the optical cell properly.
AP Ma 9003
~1990 SETZ LAQORATORIES, INC. ALLRIGHTS RESERVED.
C-1 Page t
Ihble 1. Suggestod Votumoa for Various Rangoe of CT-1 Range CT-1 (mg/L) 0.2-3.0 1.0 - 25.0 0.2-1.0 Volume CT-1 Buffer (mL) 15 10 15 Volume Dlchloroethane (mL) 10 Volume Sample (mL)
Optical Cell Size 1.0 cm 2.5 cm>><<
50cm *>>"
<<The1.0-cm cell (Code 1312) can be used with Hach spectrophotometers using a 1-cm cell adapter (Code 2776C).
- "The 2.&an cell ls the standard Hach 1-in. cell (Betz Code 2601).
<<<<>> Five centimeter cells are not available for use with the Hach photometers. Many laboratory spectrophotometers require an adapter to accommodate &cm cells. Check with the instrument manufacturer.
1.
Transfer an aliquot ofthe water sample to a separa-tory funnel (the sample). Transfer the same volume of distilled (or deionized) water to a second separa-tory funnel (the blank). Run the blank once for each set of samples tested (see Notes 1, 2, and 3).
2.
Add CT-1 Buffer Reagent to both the sample and the blank.
3.
Using a plpet, add 1, 2-dlchloroethane to both se-paratory funnels.
4.
Insert the stoppers in each ofthe separatoryfunrels.
Invert and brieflyopen the stopcock to vent the fun-nels (see Notes 4 and 5). When venting the funnels, point the tip of the funnel away from yourself and others.
5.
Shake the funnels moderately for 30 sec, vent the funnels, then allow them to stand for 10 mfn (but no longer than 15 min) ~
6.
Collect the lower layer (1, 2 - dlchloroethane) from each funnel in 5&mLbeakers leaving about 1-2 mL in the funnel. Th!s willprevent significant removal of water.
7.
Using the plastic dipper, add 2 scoops of Drying Re-agent to each beaker and stir with a glass rod for 15 sec (but no longer than 30 sec).
8.
Wait approximately 1 to 2 min (but not more than 5 min). Then carefully decant the extract offof the dry-ing reagent into an optical cell.
9.
Set the spectrophotometer at 415 nm and zero with 1, 2-dlchloroethane. Measure andrecord the absor-bance of the blank and the sample (see Note 6).
- 10. The sample absorbance minus the blank absor-bance is used to determine the concentration of CT-1 in the sample.
From a prepared calibration cutve, determine the CT-1 concentration in the sam-ple (see Calibration Curve Preparation).
- 11. Clean the cells after each measurement (see Note 7).
CALIBRATIONCURVE PREPARATION 1.
Prepare a 1000 mg/L CT-1 stock solution by accul rately weighing 1.00 g of CT-ti into 1 Lof alstillea (or deionized) water.
2.
Pipet designated volumes of the stock sotution into 1-L volumetric flasks. These are the stanaara solu-tions used in preparing a calibration cuive. Use Table 2 to make appropriate dllutlons of the stock solution for each specific app! Icatton.
3.
Followthe General Procedure using the specific so-lution volumes that have been determinea tor the application and prepare a calibration curve, Deter-mine the absorbance of a blank solution using ais-tilled (or deionized) water. This blank can oe sub-tracted from the sample absorbance or usea to zero the spectrophotometer so that the calibration curve goes through the origin. The calibration auve should be linear over the indicated ranges.
NOTES 1.
For maximum accuracy the calibration curve snould be checked by every operator using this test and shouldbe ver!fied a minimum oftwice per mcntnus-ing a freshly prepared CT-1 standard.
2.
Ablank measurement must be recorded for earn set of samples.
The blank reading may vary siigntly; however, the absolute difference between tne sarn-ple and the blank remains relatively constant.
3.
Chlorine causes a negative interference in the test.
This can be eliminated by adding 0.1 N Soaium Thiosulfate (Code 235) to the water sample before running the test. The amount added be is based on the concentration of chlorine in the system.
For a 100mL water sample containing 0.3 mg/L chlorine, add 10 drops of O.l N Sodium Thiosulfate to remove the Interference.
Page 2 C-2 AP 368 9003
~Ie 2.
DHutfone for Callbratlon Curve Prepara-tion Based on a En'olutfon Volume of 1L Concentration CT-1 Desired (mg/L)
CT-1 Stock Solu-tion Added to Make 1 L (mL) 0.2 0,4 0.6 0.8 1.0 5.0 10.0 15.0 20.0 OZ 0.4 0.6 0.8 1.0 5.0 10.0 15.0 20.0 25.0 4.
Asllghtemulslonmay formwhenusing natural water samples.
When this happens, vary step 5 of the pmcedue. Shake the hmneI for30 sec, vent It, then allow lt to stand for 5 min. Gently Invert the funnel once then allow the funnel to stand for 5 min.
5.
It Is important to vent the separatiy funnel bothbe-fore and after shaking it. Othiwise, a pressure will build up in the funnel that can cause the stopper to be forced out of the top of the ftInnel.
6.
Use caution when Insetting or removing the sample cell ln the photometer. The 1, 2-dlchloroethane can damage the call compartment.
7.
It ls imperative that the sample cells are kept clean during the running ofthe test. ItIs recommendedthat the cells are cleaned after each measurement using the following procedure:
a)
Rinse the cell three times with distilled (or deionized) water.
b)
Rinse the cell three times with methanol.
c)
Rinse the call three times with 1, 2 - dlchloro-ethane to remove methanol from the cell.
8.
This method isbasedupon Wang, L K.; Langly, D.F.
Ind. Eng. Chem., Prod. Res.
Dev., 1975, 14, 3, 210-212.
AP 388 9003
~1990 aETZ LABORATORIES,INC.ALLRIGHTS RESERVED.
C-3
TOTALANALYTICALSERVICES FOR A SAFE ENVIRONMENT I
Project Ko>>t 9117LL91 Log in Ko. t 7861 P.O.
No.
t Pend! ng Oete tAprLL 30, 1991 ANALTTICALOATA REPORT PACKAGE FOR Aqtte SUrvey Lno>>
499 Point Breete Rd.
Flenfngton, NY OBB22 ATTN:
REFt Jilt) Todd Lab Meter SAHPLE IDENTIFICATION LABORATORY NUHBER SAHPLE HATRLX SEE NEXT PAGE xv ME CERTIFY THAT THIS REPORT LS A TRUE REPORT OF RESULTS OSTALNE0 FROH OUR TESTS OF THIS HATERIAL.
RE)PE LLY SUBHIT EDr N
EHV ONHEHT L L)i PARAG )(. SHAH, Ph. 0.
ORGANIC LAB~
HANAGER OOU HEELEY ORY OLRECT R
art¹ 73469 7
))sport orl sample(s) furnished by client apprise to sample(s).
Report on sample(s) obtained by us applies only to tot sampled. (nformatxxt contaned harem e not lo be used for reproductton excepl by sceeal permisaon. Sample(s) writ be relined lor thirtydays maximum after dale of report unless spectficasy rsouested otherwise by client. In the event that there are portions or ~of sample(s) remain(ng alter Nytest has cpm(usted the reouired tests, feytest shaN have the opuon of retumeg such sample(s) to the client at the client's expense.
box 1538 a 60 seaview blvd., port washington, ny 3 <050 a(536) 625-5500 D-1
0 C
LABORATORY NUMBER SAMPLE IDENTIFICATION TYPE OF SAMPLE 7861001 7861002 7861003 LG SALT LG FRESH LP FRESH Water Water Water D-2
nytest environments DATA REPORTING QUALIFIERS U
Indicates compound was analyzed for but not detected.
Report the minimum detection limit for the sample with the U (e.g.
3.0 U ) based on necessary concentration dilution actions.
(This is not necessarily the instrument detection limit.)
The footnote should read U-Compound was analyzed for but not detected.
The number is the minimum attainable detected 1'mit for the sample.
Indicates an estimated value.
This flag is used e'ther when estimating a concentration for tentatively identif'ed compounds whe e a 3.:3. response is assumed or when the mass spectral data indicates tne presence of a co-pound that meets the identification cr'ter'a but the result is less than the specified detection limit but greate than zero (e.g.: If limit of detection is 3.0 ug/1 and a concentration of 3 ug/1 is calculated, report as 3Z.)
B This flag is used.
when the analyte is found in the blank as well as a sample.
3:t 'ndicates possible/probable blank contamination and warns t.".e data use= to take appropr'ate action.
This flag ident'fies al'a=geted compounds that were ound above the metnod detection limits.
NA This flag indicates that "he compounds are not applicable.
A Aldol condensation produc".
Note: Data on soil samples expressed on a dry weight basis.
All non-water samples are reported on soil forms. Th's includes samples whose matrix is listed as miscellaneous.
The initial and continuing calibration dates and times for the volatile fraction are listed on the BFB summary forms.
The initial and continuing calibration dates and times for the semi-volatile fractions are listed on the DFTPP summary forms.
D-3 00003
nytest environment@
Report: of Tests Water Suitability Test We find as follows:
The sample was analyzed according to Standard Methods for Examination of Water and Wastewater, 14th Ed.,
page 888.
Sample ZD:
LP FRESH Growth Ratio 1.08 Lab ED No.:
7861003 Remarks:
The sample is of acceptable suitability for the use in bacteriological testing.
A ratio greater than 3.0 would indica e
growth promoting substances present in the water.
A ratio less than 0.8 indicates the presence of growth inhibiting substances.
OO000
nytest environmental REPORT OF ANALYSZS Log Zn No.: 7861 We find as follows:
Parameter(s)
Sample Zdentification LG SALT LG FRESH LP FRESH Method (7861001)
(7861002)
(7861003)
Blank pHSalinity, ppt'.66 29 '
NA
,<1 I
Results in mq(1:
Ammonia Nitrogen Bromide Fluoride Zodine Nitrate Phosphate Residual Chlorine Sulfide
'ulfate Total Cyanide Total Suspended Solids
- 0. 130 40.8 0.576
<1
<0.04
<0.04
<0.1
<0 ~ 10 1910
<0 F 01 3
0.22
<0. 1
<0 '
<0.05
<1.0
<0.02
<1
<0.04
<0.04
<0.1
<0.10
<1
<0.01
<1 ppt
~ parts per thousand D-5 00005
nytest environments REPORT OF ANALYSIS Log In No.:7861 We find as follows:
Results in mg/1:
Parameter(s)
Sample Identification LG SALT LG FRESH LP FRESH Method (7861001)
(7861002)
(7861003)
Blank Aluminum Arsenic Beryllium Boron Cadmium calcium Chromium Cobalt Copper Iron Lead Magnesium Manganese Mercury Molybenum Nickel Potassium Selenium Silver Zinc Total Organ ic Carbon
<0.2
<0.010
<0.005 3
3
<0.005 271
<0.010
<0.05
<0.025
<0.050
<0.050 932
<0.015
<0.0002 0-16
<0.040 348
<0.010
<0.010
<0.020 20 '
11.3
<0.010
<0.005
<0.025
<0.050
<0.0002
<0.040
<0.020
<0.2
<0.010
<0.005
<0-3.
<0.005
<1 ~ 0
<0. 010
<0.05
<0.025
<0.050
<0.050
<1.0
<0.015
<0.0002
<0.010
<0.040
<1.0
<0.010
<0.010
<0.020
<1 '
00006
1ST NYTEST EHVIRONHENTAL INC.
TCL PESTICIOE/PCS ORGANICS ANALYSIS OATA SHEET SAHPLE HATRIX: llATER CONC. LEVEL: LOM EXTRACTION OATE: 4/9/91 ANALYSIS OATE: 4/18/91 SAHPLE '10 LAS SANPLE 10t OIL FACTOR:
X HOISTVRE:
VG/L LG SALT 7861001 1.00 HA CHPO 0 CAS Number PEST ICIOE/PCS CCSIPOJHO 1
[ 319-84.6 2
I 319 85 7 3
I 319868 4
I 58 89 9 5
i 76-44-S 6
I 309-00-2 7
I 1024.57.3 8
I 959 98 8 9
I 60 57 1
10
/ 72.55-9 11 I 70 20 8 12 I 33213 65 9 13 I 72 54 8 14 I 1031-07-8 15 I 50 29 3 16
] 5349C-70.5 17
( 72.C3.5 18 I 57 74 9 19 I 8001-35-2 20 I 12674-11.2 21 I
11104 28 2 22
) 11141.16.5 23 i 53C69.21-9 24
[ 12672.29-6 25 I 11097.69.1 26 I 11096 82-5 I
[ Alpha.BHC f Beta-BHC i Delta.BHC
[ Ganma-BHC(Lindane)
] Heptachlor
( Aldrin
[ Heptachlor Epoxide
( Endosulfan I
( Oieldrin
) 4,4'-OOE f Endrin
( Endosulfan II t 4,4-000
( Endosulfan Sulfate I C,4'-OOT
) Endrin Ketone
( Hethoxychlor
( Chlordane Toxaphene
( Aroclor-1016
( Aroclor-1221
) Aroclor' 1232
) Aroclor-1242
( Aroclor-1ZCS t Aroclor-125C
[ Aroclor-1260 I
I I
I I
I I
I I
I I
I I
I I
I I
I I
I I
I I
I I
I I
0.050 U.
0-050 U.
0.050 U.
0.050 V.
0 050 U.
0.050 U.
0.050 U.
0.050 U.
0.100 U.
0.100 U.
0.100 U.
0.100 U.
0.100 U.
0.100 U.
0-100 V.
0.100 U.
0.500 U.
0.500 U.
1.000 U.
0.500 U.
0.500 U.
0.500 U.
0.500 U.
0.500 U.
1.000 U.
1.000 U.
D-7 00007
ORGANOPHOSPHOROUS PESTICZDES Lcg in No.:
7861 Sample ZD No.:
LG SALT Lab ZD No.:
7861001 Parameters (ug/1)
Detection Limit (ug/1)
Found Vapona (Dichlorvos)
Phorate Diasinon Naled (Dibron)
Dementon - S Disulfoton Ronnel Chlorpyrifos (Dursban)
Bolstar (Sulprofos )
Stirophos Fensu1f othion Azinphos methyl (Guthion) 0.2 0.2 0.6 0.2 0.2 0 '
0 '
0.3 0 '
2.0 5.0 1 '
ND ND ND ND Coumaphos 1.5 Malathion 0.3 Parathion 0.3 ND ~ Not Detected 0-8 on008
1OT NYTEST EHVIROHHENTAL INC TCL PESTICIDE/PCS ORGAHICS ANALYSiS OATA SHEET SAHPLE HATRIX: HATER CONC. LEVEL: LOM EXTRACTIOH OATEt 4/9/91 ANALYSIS DATE: 4/19/91 SAHPLE IO:
LAS SAHPLE ID:
OIL FACTOR:
I NOISTUREt UO/L PBLKI A040005 1.00 NA CHPO ¹ CAS Nunber PEST ICIOE/PCS CCHPtRIHD 1
( 319.84-6 2
( 319.85-7 I 319 86 8 4
( 58.89-9 5
( 76-44.&
6 I 309.00-2 7
I 1024.57 3
8
( 959-98 8 9
( 60-57-1 10
( 72-55-9 11 I 70-20-8 12 I 33213-65.9 13 I 72-54-8 14
( 1031-07.8 15 I 50.29-3 16
( 53C9C-70.5 17 I 72 43 18 I 57.74-9 19 I 8001-35-2 20 I 12674-11-2 21 I 11104-28-2 22 I 11141-'16.5 23
( 53469-21-9 24
( 12672.29.6 25
( 11097.69-1 26
( 11096-82-5 I
( Alpha BHC
( Beta.BHC
( Oelta.BHC
( aaaea.BHC(Lindane)
( Heptachlor
( Aldrin
( Heptachlor Epoxide
( Endosulfan I
( Oieldrin
( C,4'-OOE
( Endrin
( Endosultan tt
( C,4-000
( Endosulfan Sulfate
( 4,4'-ODT
( Endrin Ketone
( Hethoxychtor
( Chlordane
( Toxaphene
( Aroclor-1016
( Aroclor-1221
( Aroclor-1232
( Aroctor-1242
( Aroclor-1248
( Aroctor-1254
( Aroclor-1260 0.050 U.
0.050 U, 0.050 U.
0.050 U.
0.050 U.
o.oso u.
0.050 U.
I O.oSo u.
0.100 U.
0.100 U.
I 0.100 U.
I 0.100 U.
0.100 U.
I 0.100 U.
0.100 U.
I
- o. loo u.
o.sao u.
o.soo u.
1.000 U.
I o.sao u.
o.soo u.
Q.saa U.
o.soo u.
(
0.5QQ U.
1.000 U.
I 1.0oo u.
D-9