1*
*
TNO Environmental and Energy Research
*
c
'Z3?ai TNO Instituteof Environmental Sciences Schoemakerstraat 97 P.O. Box 6011 2600 JA Delft The Netherlands Fax+31 15 61 68 12 Phone+31 15 69 69 00
TP'O-repot IMW-R 92/348a
RijkswaterstaatiRlZA Rljksintltui.it voor lngrsaf Zcetwate.rbchier en Afvaiwmrbehafldelirig Documentatie losi bus 7 5270 AA Ldysta
The acute toxicity of praseodymium chioride heptahydrate to the marine copepod Acartia tonsa
Authors : Dr C.T. Bowmer Ms J.A. Kauffman-van Bommel Ms C. Degeling
TNO Study no. : IMW-92-0069-01
Date : June4, 1993
Order no. : 51795
Al] rights reserved. No part of this publication may be reproduced and/or published by print, photoprint, microfilm or any other means without the previous written consent of TNO. In case this report was drafted on instructions, the rights and obligations ot contracting parties are subject to either the Standard Conditions for Research Instructions given to TNO', or the relevant agreement conciuded between the contracting parties. Submitting the report for inspection to parties who have a direct interest is permitted.
Sponsor: Rijksinstituut voor integraal zoetwaterbeheer en afvalwaterbehandeling (RIZA) Postbus 17 8200 M LELYSTAD The Netherlands
Approved by: Dr R.J. Dortland
Version: AcToxCrustAtonsall 992/10/19
Number of pages: 38
TNO
Netherlands organization for applied scierstific research Based on the necessity for a suataijaYe development of 'wciety, TNO Environmental and Enen1v Research aims al contributing, through research and adv os adequate erivironmental management, rational energy c,nsumption and the proper management and uae of subsur'ace natural reaourcea.
The Standard Conditions for Research Instructions given to TNO, as tiled af the Registry of the District Court and the Chamber of Commerce in The Hague shal apply to all inatructions given to TNO.
IMW-R 921348a
page 2 of 38
RA92348aI MWjt
CONTENTS
page TITLE
1
CONTENTS
2
CONFIDENTIALITY STATEMENT
3
GLP COMPLIANCE STATEMENT
4
CONTRIBUTING PERSONNEL
5
QUALITY ASSURANCE STATEMENT
6
SUMMARY
7
1. INTRODUCTION
8
2. MATERIALS AND METHOD 2.1 Test sub stance 2.2 Test organism 2.3 Dilution water quality 2.4 Test method 2.5 Treatment of the resuits 2.6 Chemical analysis
9 9 9 9 10 11 11
3. RESULTS 3.1 Environmental conditions 3.2 Chemical analysis 3.3 Mortality and other effects
13 13 14 14
4. REFERENCES
16
5. RETENTION OF RECORDS AND SAMPLES
17
6. DEVIATIONS FROM THE PROTOCOL
17
ANNEX A PRODUCT INFORMATION ON PRASEODYMIUM CHLORIDE 18 HEPTAHYDRATE ANNEX B INDIVIDUAL MORTALITY, CONDITION, pH, OXYGEN 19 CONCENTRATION AND TEMPERATURE MEASUREMENTS ANNEX C ESTIMATION OF THE LC50 AND ITS CONFIDENCE INTER VAL 36 ANNEX D CHEMICAL ANALYSIS
37
IMW-R 92/348a
page 3 of 38
RA92348aI MWjt
CON FIDENTIALITY STATEMENT
This report contains confidential and proprietary information of the Rijksinstituut voor integraal zoetwaterbeheer en afvalwaterbehandeling (RIZA) which must not be disciosed to anyone except the employees of RIZA, without the express and written approval of RIZA.
zw
IMW-R 92/348a
page 4 of 38
RA92348aI MWjt
GLP COMPLIANCE STATEMENT
'1, the undersigned, hereby declare that the work to which this report refers was performed under my supervision according to the procedures herein described. To the best of my knowledge this report provides an accurate record of the resuits obtained. This study was carried out in compliance with the OECD code of Good Laboratory Practice.'
Dr C.T. Bowmer Study Director
Date:
2 ƒ (~ <~ 13
IMW-R 921348a
page 5 of 38
RA92348aIMWjt
CONTRIBUTING PERSONNEL
Ms J.A. Kauffman-van Bommel Technician
Date:
Department of Biology
Ms C. Degeling Trainee technician Department of Biology
Date:
Date: Study Supervisor
Head Department of Biology
7
z-
93
page 6 of 38
IMW-R92/348a MAB
Report no.: IMW-R92/348a Quality Assurance Unit-IMW Study no. : IMW-92-0069-01 P. 0. Box 6011 2600 JA DELFT
QIJALITY ASSURANCE STATEMENT STUDY TITLE: The acute toxicity of praseodymium chioride heptahydrate to the marine copepod Acartia tonsa REPORT DATE: June 4, 1993 The following inspections relevant to this study have been carried out by the Quality Assurance Unit of the TNO Institute of Environmental Sciences (1MW), P. 0. Box 6011, 2600 JA Delft, the Netherlands.
Type of inspection
Date and nuniber of inspections
Date of report to Study Director
experimental phase
September 22, 1992 (1) October 8, 1992 (1)
September 23, 1992 October 8, 1992
September 29, 1992 (1) November 2, 1992 (1)
September 29, 1992 November 2, 1992
December 7, 1992 (1) January 6, 1993 (1) June 9, 1993 (1)
December 8, 1992 January 6, 1993 June 9, 1993
experimental phase of equivalent* studies:
report audit:
Any serious deviations were reported to management at the same time as the report to the study director; any other, less serious deviations were reported to management upon receipt of the reply from the Study Director. * This type of study is carried out regularly by the personnel responsible for the present study; these dates refer to inspections of such equivalent studies with other test substances. 1, the undersigned, hereby declare that to the best of my knowledge this report provides an accurate record of the resuits obtained in this study.
/-
M. A. Bikker / Quality Assurance Officer Date:
/3/3
IMW-R 921348a
page 7 of 38
RA92348aIMWjt
SUMMARY
The acute toxicity of praseodymium chioride heptahydrate to Acartia tonsa (Copepoda: Crustacea) was determined according to a proposed ISO Guideline (ref. 1) and a PARCOM protocol (ref. 2), carried out under the OECD principles of Good Laboratory Practice (ref. 3). The test was carried Out under static conditions without replacement of the test solutions, the exposure time being 48 hours. The dosed amounts tested were 0, 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg of praseodymium chioride heptahydrate (in the form received from the supplier, expressed as praseodymium) per litre of ca. 28%c natura! seawater. The aqueous solubility of the test substance appeared to be good (visually assessed at the concentrations tested). All concentrations from 10 mg.1 -1 and higher appeared initially to be a milky white colour. However, within a short space of time a layer of flock was evident on the bottom of the test vessels. The resuits of the test based on dosed amounts of praseodymium were (confidence interval between brackets): 24h LC50 : 76.0 mgk' (14.6-396 mg.1') 48hLC50 : 19.3 mgk 1 (11.0-33.9mg.1 1 ) 48hLC100 48h NOLC 48hNOBC 1 )
:
>32
:
:
mg.1'
3.2 mg1 1 1.8
mg.1 1
Analyses of samples of selected test so!utions taken at the start and the end of the test indicated disso!ved praseodymium concentrations of 1 to 6% of the nominal dosed amounts. The range of variation of the pH and 02 conditions observed in this test are un!ikely to have adversely affected the re!iabi!ity of the resuits of this test.
1) Swimming behaviour/mobility.
zw
IMW-R 92/348a
page 8 of 38
RA92348aI MWjt
1. INTRODUCTION
The acute toxicity of praseodymium chioride heptahydrate to the copepod crustacean
Acartia tonsa was determined according to a proposed ISO Guideline (ref. 1) and a PARCOM protocol (ref. 2). In conformity with the OECD principles of Good Laboratory Practice (ref. 3) the test was carried out under static conditions without replacement of the test solutions, the exposure time being 48 h. The objective of the study is to determine the 48h LC50 (lethal concentration, 50%) and the 48h NOEC (no-observed-effect-concentration) of the test substance (as praseodymium) with the marine copepod Acartia tonsa. Relevant dates for the test were: Protocol signed by Study Director on: September 21, 1992. Period of range-finding test: September 22-24, 1992. Period of final test: October 7-9, 1992. In this 'a' version of report R921348 the draft analytical chemistry data in Annex D has been replaced by the definitive data. Through the above changes small alterations in the chemical data occurred and have been corrected in the report.
IMW-R 921348a
page 9 of 38
RA92348a!MWjt
2. MATERIALS AND METHOD
2.1 Test substance The substance examined was praseodymium chioride heptahydrate, a solid consisting of "green crystals and small chunks". The test substance was received on September 11, 1992 in a 15 mi plastic jar with a black screw-cap, labeiled: "02427 CX DX; Aidrich 20.413-7; praseodymium chioride heptahydrate; 99.99% Irritant! Hygroscopic!; F.W. 373.37; m.p. 115° d 2.250". The sample was stored at room temperature in a ciosed cupboard. The composition and properties of the test substance as specified by the supplier are recorded in Annex A.
2.2 Test organism The test animal was the marine crustacean Acartia tonsa (Copepoda: Crustacea), cultured in the laboratory under standard conditions according to the relevant Standard Operating Procedure. The animals used in this test were copepodites (subaduits) between 6 and 8 days old.
2.3 Dilution water quality The dilution water used was natural seawater, kept in storage tanks in the laboratory. The stock is regularly supplied with seawater, originating from the Eastern Scheidt, a virtualiy unpoliuted area in the Netherlands. Upon arrival the seawater is checked for the concentration of voiatiie organics, TOC and minerai oil; it is only used if concentrations of these compounds are found to be acceptably low. In addition, a 96 hour bioiogical test with
young specimens of either Chaetogammarus marinus or Mysidopsis bahia is carried
Out.
Before entering the storage tanks, the water is filtered through a sandfiiter. Furthermore, immediately before use, the water is filtered over a carbon filter and a 5 pm membrane filter. The salinity of the seawater is normally ca. 33%. For this test with Acartia tonsa it was diluted to ca. 28%o with distilled water.
zw
IMW-R 921348a
page 10 of 38
RA92348aI MWjt
2.4 Test method The 48 h test was carried Out according to a protocol which has been developed from a proposed ISO Guideline (ref. 1) and a PARCOM protocol (ref. 2). The test was conducted in accordance with the OECD principles of Good Laboratory Practice (ref. 3). The duration of the test was 48 h and appropriate test concentrations were determined in a preliminary range-finding test. A quantity of the test substance (6629 mg) equivalent to 2500 mg of praseodymium was accurately weighed out and dissolved in 250 ml of 'Milli-Q' water to achieve a stock solution of 10 gt 1 . Aliquots of 0.1, 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 ml were made up to 1 litre with ca. 28%c natural seawater to achieve test concentrations of 1.0, 1.8, 3.2, 5.6, 10, 18 and 32 mg praseodymium.1 1 . Natural seawater was used as a control medium. About 10 ml of each of these test solutions was placed in each of ten small glass test vessels (scintillation vials, coded 1 to 10). Natural seawater (ca. 28%o) was used as a control solution. The control and test solutions were not replaced. However, after 24 and 48 h exposure, the numbers of living and dead animals were counted and the condition of the survivors was visually estimated. The pH and oxygen concentration of the clean control and test solutions were measured before introduction to the test vessels. The pH and oxygen concentration were measured in all replicates where mortality had occurred after 24 and 48 hours. Regardless of mortality, these parameters were measured in one replicate of the control and each test concentration after 48 h exposure. The test was carried out in a temperature- and light-controlled room at 20 ± 1°C and a 16 h light-8 h dark regime with two transition periods of 0.5 h of twilight. The temperature of the control solution was measured at the beginning and end of the test and the salinity of the dilution water was measured at the start of the test. The test animals were not fed during the test.
IMW-R 92/348a
page 11 of 38
RA92348aIMWjt
2.5 Treatment of the resuits
LC5O value The effect of a test substance on the mortality of animals is expressed by a quantity denoted as the LC50 (= Lethal Concentration, 50%), i.e. the exposure concentration of the substance which would prove lethal to 50% of an infinite population of the exposed animals. For these calculations the combined mortality data per test concentration were used. The 24h and 48h LC50 values and their confidence intervaJ were calculated by means of a parametric model developed by Kooijman (ref. 4). A summary of this method is given in Annex C.
NOLC value The 48h 'no Qbserved lethal çoncentration' (NOLC value) is the highest concentration tested showing no mortality (= lethality) throughout the exposure time. The 48h NOLC value is estimated by comparing effects with regard to mortality/survival of the exposed animals with those of the control animals.
NOEC value The 48h 'o Qbserved effect Qoncentration' (NOEC) is the highest concentration test in which at least 90% of the animals survived for 48 hours and did not show any signs of harm caused by the test substance. LC100 value The LC100 value is the lowest concentration tested showing total mortality.
2.6 Chemical analysis
The actual concentrations of soluble praseodymium in the test media was deterniined by chemical analysis. Samples of ca. 100 ml were taken at t = 0 h from the control medium and the dosed amounts of 1.0, 5.6 and 32 mg.l' as well as from the same "spent" media at the end of the test.
Time
IMW-R 92!348a
page 12 of 38
RA92348aI MWjt
All samples were filtered through a 0.45 tm filter. The samples were transferred to polyethylene bottles and then a 100 jiJ aliquot of 70% HNO3 was added to each sample for conservation. The samples were stored in a refrigerator until they were transferred to the department of Analytical Chemistry. The samples were analysed by Inductively Coupled Plasma - Atomic Emission Spectrometry (ICP-AES). Details on the chernical analysis are given in Annex D.
IMW-R 92/348a
page 13 of 38
RA92348aI MWjt
3. RESULTS
The aqueous solubility of the test substance appeared to be good (visually assessed at the concentrations tested). All concentrations from 10 mg.1' and higher appeared initially to be a milky white colour. However, within a short space of time a layer of flock was evident on the bottom of the test vessels.
3.1 Environmental conditions The following environmental parameters were measured in the test solutions. pH The pH values of all test and control solutions during the test period varied between 7.1 and 8.1. The pH values measured are recorded in Annex B, Table B2. Oxygen concentration The oxygen concentration on all test and control solutions during the test period was greater than 7.3 mg.1 -1 . The oxygen concentrations measured are recorded in Annex B, TableB3. Salinity The saJinity of the dilution water used for this test was 28.7%. Temperature The temperature as measured in the control solution, varied between 19.8 and 20.8°C (start and finish); these values measured are recorded in Annex B, Table B4. The range of variation of the above parameters is unlikely to have affected the reliability of the resuits of this test.
Me
zw
IMW-R 921348a
page 14 of 38
RA92348aIMWjt
3.2 Chemical analysis
The resuits of the chemical analysis are given in Table 1; these data are derived from Annex D. Table 1 Chemical analysis of soluble praseodymium during the acute toxicity test with Acartia tonsa. The purisy of praseodyinium chioride heptahydrate was 99.99%. Samples just after dosing (t = 0 h)
Dosed amount of praseodymium (m9.1 1
)
Pr (m9.1 1
)
% of dosed
Samples of spent solutions (t = 48 h) Pr (m9.1 1
)
% of dosed
% of t = 0 h
amount
amount
0
<0.1
<0.1
1.0
<0.1
<0.1
5.6
0.1
2
0.2
4
200
2.0
6
0.3
1
15
32
The actual concentrations of soluble praseodymium were far below the dosed arnounts. Just after dosing 2% and 6% of the dosed amounts of 5.6 and 32 mg Pr.1 1 were measured respectively. After 48 h exposure 200% and 15% of the measured concentrations just after dosing was stili present respectively. These resuits suggest the complexation of praseodymium in seawater.
3.3 Mortality and other effects
The LCSO, LC100, NOLC and NOEC values are recorded in Tabie 2 and data on survival in Table 3. Details on mortality and condition are presented in Annex B, Tabie Bi. The slope of the concentration-effect curve (the inverse of parameter 3 of the model, see Annex C) was 1.3 with a 95% confidence interval of 0.87 to 2.9. The resuits of the chemical anaiysis do not aliow the extrapolation of the LC50 vaiues in terms of measured concentrations of solubie Pr. The actual concentrations of solubie Pr, however, were far below the dosed amounts.
IMW-R 921348a
page 15 of 38
RA92348aI MWjt
Table 1 Resuits of the test with praseodymiumchloride heptahydrate and Acartia tonsa (confidence interval between brackets). Parameter
Dosed amount
Effect
mg Pr.1 1
(h = hours)
24h LC50
mortality
76.0 (14.6-396)
48h LC50
mortality
19.3 (11.0-33.9)
48hLC100
mortality
48h NOLC
mortality
3.2
48h NOEC
swimming behaviour / mobility
1.8
>32
Table 3 Summary data on survival1 ). Time
1)
Dosed amount(mg_Pr.1 1 )________
1
1.8
3.2
5.6
10
10
10
10
10
10
10
10
10
10
10
9
8
9
9
10
10
10
10
7
6
6
3
(h)
0
0
10
24 48
1.0
For details on condition seethe raw data in Annex B, Table Bi.
AIMVA-
10
18
32
IMW-R92/348a
page 16 of 38
RA92348aI MWjt
4. REFERENCES
Water Quality - Determination of acute lethal toxicity to marine copepods (Copepoda: Crustacea), UK proposal to ISO, TC147ISC5IWG2 (November, 1990). Bjornstad, E. Paris Commission: Ring-test protocol, Acute toxicity to the marine copepod Acartia tonsa, Water Quality Institute, Horsholm, Denmark (January, 1991). Good Laboratory Practice in the testing of chemicals. Organization for Economic Co-operation and Development, Paris (1982). Kooijman, S.A.L.M. (1981). Parametric analyses of mortality rates in bioassays. Water Res. 15,pp. 107-119.
zw
IMW-R92/348a
page 170f 38
RA92348aIMWjt
RETENTION OF RECORDS AND SAMPLES
All the data generated and all other records and information relevant to the quality and integrity of the study have been filed under the study reference IMW-92-0069-0 1 in the archives of the TNO Institute of Environmental Sciences, Schoemakerstraat 97, 2628 VK Delft, The Netherlands. These records will be retained for a period of at least ten years after the cover date of this report. A sample of the test substance has been deposited under the reference IMW-92-0069-A in the sample archives of the TNO Institute of Environmental Sciences at the same address; this sample will be stored for a period of at least 10 years.
DEVIATION FROM THE PROTOCOL
• The name of the test substance was praseodymium chloride heptahydrate and not 'praesodymium chloride hexahydrate'. • The test animals were between 6 and 8 days old at the start of the test and not between 6 and 7 days as stated in the protocol.
IMW-R 92/348a
page 18 of 38
RA92348aIMWjt
ANNEX A PRODUCT INFORMATION ON PRASEODYMIUM CHLORIDE HEPTAHYDRATE
aldrlch chemical co.. chemtsts helping chemists in research & iridusry
PRODUCT INFORMATION PRODUCT NUMBER: 20413-7
t.LQT NUMBER: 02427CX
PRODUCT NAME: PRASEODYMIUM CHLORIDE HEPTAHYDRATE, 99.99 FORMULA: PRCL3.7H20
FORMULA WEIGHT: 373.37
APPEARANCE
GREEN CRYSTALS AND SMALL CHUNS
X—RAY DIFFRACTIOP4 PATTERN
CONFORMS TO STANDARD PATTERN
TITRATION
37.8% PR (COMPLEXOMETRIC)
TRACE ANALYSIS, ICP
RARE EARTH ELE ME NT S YB 30 PPM DV 25 PPM
OTHER EL EM EN T S PPM CA 10 PPM Nl 6 MB 0.7 PPM
'.LORICH war.nt, that tts products conform to the information contsined In this nd other Aidr$ch pubilcations. Purchaser must determine the suitability of the )roduct lor t, particuiar usa. S.* reverse dde of invoice or paclçinQ slip for addt• onal term, and conditions of 5.1..
•
AIdrch Chemical Company RUTH WANTZ APRIL 24, 1990
akirich chemical co. P.C. Box 355,
MUwiuk, WIscen*In 53201 USA e (414)273.3850
AWAO-
zw
IMW-R 921348a
page 19of38
RA92348aIMWjt
ANNEX B INDIVIDUAL MORTALITY, CONDITION, pH, OXYGEN CONCENTRATION AND TEMPERATURE MEASUREMENTS (copy of raw data)
Table Bi Data on mortality and condition of the test animals.
INSTITUUT VOOR MIUEUWETENSCHAPPEN TNO AFDELING BIOLOGIE Proeicode Forrwtier : IMWB/FM/4022 Voor Registratie overlevende dieren, tEn oonditie en ae opmerkingen bij de acult sen statische toets met Acartia tonsa (481t); t dier per pioelvat Behooil bij : Werkvoorscliatl IMWBISM/4020
Datum TIjdsfl
o7o
10 5
Proeltijd (h) 0 cond
Concentratie mg.r'ig.r',vo&%
Paraat
1 2 3 4 5 6 7 8 9 10 __
c
24 cond.
lo 5
48
Conditie dieren: a - ncmaal b - ge4i aan cortole haag, versuft
a. D.m
I6 fïY
c
Ck
a - nonnal b - eial to corimi
IMW-R 92/348a
page 20 of 38
RA92348aIMWjt
INSTITUUT VOOR MILIEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE Proef Code t6 «Tr
Fonrvt,er IMWB/FM/4022 Voor Regstralre overlevende dieren, lijn oorwitie en afle opmerkingen bij de acute seni-statische toets met Acaflia tonsa (48h); t dier per proel val Behoort bij = Werkvoorschnd IMWB/SM/4020
Datum Tijdstip Proetijd (h) 22
Concenmg.r1 4rç4ireM 1
2
3
4 $ 6
0 cond.
o 5
-
24
cofl(j
48 cclxi
Conditie dieren: a - normaal b - geliØ aan oentrole traag, versult : r9
erge
d.kç i,lWT)C. 0.11
normad - ecpalIocortol dead
Paraat 7
8
9
10
IMW-R 921348a
page 21 of 38
RA92348aI MWjt
INSTITUUT VOOR MILIELJWETENSCIiAPPEN TNO AFDELING BIOLOGIE
1
Proef code
Fomijtier : IMWBIFM/4022 Voor Registratie overlevende dieren, tuin conditie en ale opmerkingen bij de acute seni-statische toets met Acaffia tonsa (48h); t dier per proetvat Behoort bij Wedcvoorschritt IMW13/SM14020
Datum Tijdstip PToettijd (h) gz -----2.5
0 co4ld.
24 %o5
cood. 48
Concentratie 1
2
41
-.
3
mg.r' J 1 vo&% (.7
4 5 6
Mie..gBieogi.. øaie.1,.D.e
8
9
b
t,
L
cond.
Conditie dieren: a - formaat b - geliØ aan coroêe traal ver
Paraat 7
- eciat to coatml dead
10
page 22 of 38
IMW-R 92/348a RA92348aIMWjt
INSTITUUT VOOR MILIEUWETENSCHAPPEN TNO AFDELING BIOLOGIE
Proef code
Fomsiher IMWB/FM/4022 Voor RestraUe overlevende dieren. Ivin conditie en aae opmetkalgen bil de aJte setTi-atische toets met Acattia tonsa (48h) t dier per proefvat Behoort bij Werkvoorschrifl 1MWB/SM/4020
tijd (h) ------0 cond.
'2
1 2 3 4 5 6 7 8 9 10
b
24 cond
6
L
Z
cond.
Conditie dlersn: a - normaal b - ~ aan coritrole onbewegeç traag, versuft
-.
Paraat
Concentratie mg.r'grvofk
Datum Ti)dst
-rom - equal to control - rwronte, -dead
lW.T?. O
zw
c
IMW-R 921348a
page 23 of 38
RA92348aIMWjt
INSTITUUT VOOR MIUEUWETENSCHAPPEN TNO AFDELING BIOLOGIE
Proef code
Ri RT,
Forn*jtier IMWB/FM14022 Voor Regis*ratle overlevende dieren, hun conditie en afle opmerkingen biÈ de acute semi-alische toets met Acafla tonsa (48h); t dier per proefvat Behoort bij Werkvoorschritt IMWBISW4020
Datum lljdstIl
C7
U) 3S
Proeftijd (h)
48 cond.
Conditie dieren: a - normaal b - geI aan contiole gekØi. traag. :
v92C501
Paraat
5.4
1 2 3 4 5 6 7 8 9 10
0 cond.
24 cond.
15
Concentratie mg-t4ig-t'/veM
PIW.TNO. 0.m
c
6 6
6 6
6 6
noffrsd - ecp.ral to control daad
-
h dwged
6
Jk
MW-R 92/348a
page 24 of 38
RA92348aIMWjt
INSTITUUT VOOR MIUEUWETENSCIIAPPEN TNO AFDELING BIOLOGIE
1
proelcode
Fornulier IMWB/FM/4022 Vooi' Restratie overfevende dieren, hun conditie en afle opmerkrngen bij de acute seni-statische toets met Acan'ia lonsa (48h) t dier per proefvat Behoort bij Werkvoorsctwitt IMWB/SM/4020
Datum TIjdstl
Proef. tld
1
2
3
4 5 6
b
24 con
c
d 6
..
'
Conditie dlern: a - normaal b - gel*aancoritrole c = onbewe^ traag. verstit
Bieoe.. effiC.
7
8
9 10
.
Cio
1,9
Paraat
ConCentratIe mg.rlJ4,e%
0 cond.
to 2.5
Tii:J
0.0
-normal = eiaI to control dead
6
.
—
IMW-R 92/348a
page 25 of 38
RA92348aI MWjt
INSTITUUT VOOR MILIEUWETENSCI-IAPPEN TNO AFDELING BIOLOGIE Proetcode Fomulier IMWB/FMJ4022 Voor Res*raie overlevende dieren, hun conditie en afle oprnerlungen bij de aCut3 seni-atische toets met Acan'ia tonsa (48h); t dier per proetval Behoo,t bi : Wettvoorsctirut IMWB/SM/4020
Datum 'n)dstlp Proeftijd
COnCentratie mg.F'ig!l/voL% &u
Paraat
1 2 3 4 5 6 7 8 9 10 '
0 cond.
24
comi.
Q
6
48 coixi
Conditie dieren: a - formaat b = geliaanconttole Iraa. versdt
e_. eiwn o.*
- equaltoax*ml dead
L
-
IMW-R 921348a
page 26 of 38
RA92348aI MWjt
INSTITUUT VOOR UILIEUWETENSCHAPPEN TNO AFDELING BIOLOGIE Proef code I?(
Forimlier : IMWB/FM/4022 Voor Registratie ovedevende dieren, hun conditie en ale opmeilungen bij de acule sers-stalische toets met Acartla tonsa (48h); 1 dier per proelvat Behoort bij Wedcvoorschritl IMWB/SM/4020
Datum
td
-S'
'
=
1o25
Concentratie mg.r 1 1
2
3
t-1Nch%
4 5 6
0 cond.
Paraat
sz 7 8
9
10
'
24 corst.
b
48 cond.
C
Conditie dieren: a - normaal b - geIiaancor*ro4e
Fi7
b
=t
- equalt000nitrol - irimobile, s,ggish. diuged - dead
&rU
ii
c
-
IMW-R 92!348a
page 27 of 38
RA92348aIMWjt
Table B2 pH measurements.
INSTITUUT VOOR MUJEUWETENSCHAPPEN TNO AFDELING BIOLOGIE
Prosicode Pr?
Fomulier : IMWB.'FM/4023 Voor Regietie ., pH.waarden Ii vese i Qeblutse oplos*.en Ii de acute sei'sche toets met Asta tonea (48 h); 1 er per soetvet Behoort bij We,kvoordt MTB.'SM/4020
ccntsetI.m9J4*4.r1Ns% (h) -
?.1
1
2 3 4 5 6 7 8 9 10
c S C S C S C S C S C S C S C $ C S C S
eij~
.
24
• 5,'
r
-
. . . . .
tijd (ii)
. .
. . . .
•
.
. .
•
•
1 2 3 4 5 6 7 8 9 10 C $ C $ C $ C S C S C S C S C S C S C S
0 24
-
,. . . . . . . .
. . •
. .
.
48
c: dei (schone) soete4Js19 s: upent (wie) betscØoseNig
-
-
zw
.
.
. .
-
IMW-R 921348a
page 28 of 38
RA92348a1 MWjt
RISTTTUUT VOOR MUJEUWETENSCHAPPEN fl10 AFDEUHG WOLOGIE Fonmjlier : IMWBIFW4O23 ________ Voor Retjie val pH-waardon Ii ves en gebt*ts oesen in de ~e se-etisdle toets met Acailia tonsa (48 h) 1 def per Betooa bij Wwtcvoorsctdt MTBISW4020
an TIJdstq
Pro&code: c
Paraat
ConcsntraUe mg.r'4tLr'NOLS „8
Prost. tijd (h)
1 2 3 4 5 6 7 8 9 10 C S C $ C S C S C $ C S C S C $ C S C S
%
oBo
0 24
- -
a...
48
qz.
Paraat
Col*lalism9i 14igJtvsL% 3 •
tijd (t)
1 2 3 4 5 6 7 8 9 10
-
C $ C S C S C S C $ C S C S C S C $ C S
0 24 48
-
-
-
-
c deal (schone) to ik s: apset (wie) IoetsopIosng
-
- -
-
-
-
IMW-R 921348a
page 29 of 38
RA92348aIMWJt
INSTITUUT VOOR UILIEUWETENSCHAPPEPI TNO AFDEUNG BIOLOGIE FomjIier : IMWBFW4023 voor : Registretie van pH-waarden ki verse en gebde optoss1gefl in de aae sen-s*aisde toets met Acatia tonsa (48 b); 1 dierperproefvat Behoo,t bij WeikvoosscJ't1 MTB1SM/4020
Da$isfl
_.% CoiicsntiitfS ing.I44J 1 ?vat
Tijmo Proef-
tijd (h)
Paraat
Ç
1 2 3 4 5 6 7 8 9 10 C $ C S C S C S C $ C S C $ C
_•3_ 0
$ C $ C S
3
24 !2,2
Proetcodo 101 t Z C A1
0
0
48 -
Concantreti. mg.r'4+r1ivsL%
)atum TI~IP Proeftijd (h)
-
Paraat
to
1 2 3 4 5 6 7 8 9 10 C S C 5 C $ C $ C S C S C S C S C S C
24 48
9,o
-
-
c dea' (ecIoe) toefptoeiaiu ipent (vul.) bopkieii151
•:
è.
,'
mw
$
-
IMW-R 92/348a
page 30 of 38
RA92348aIMWjt
INSTITUUT VOOR MILIEUWETENSCHAPPEN TNO AFDELING BIOLOGIE
:
Proetoode:
Fomulier IMVVB~4= Voor Re9istraie ven pH.waaden 1s verse en gebludde cØosskigen Ii de ate wn~~ toets met Aca,tia Ioreta (48 h); 1 dIe( pef Behoort bij Werkvoorsctwlt MTBfSW4020
DMM T~IP 'P~ qd (h)
ticsf1retIS 1
mg.t'gi'/vot% 9
2 3 4 5 6
7
AT
rt'
-
Parsaf 8 9 10
C $ C S c S C S C $ C S C S C S C S C S
_133_
°
.
24
-
.
.
_
J'NoL%
. .
7
1 2 3 4 5 6
.
24
.
6'
.
•
•
.
•
- (schone)
•
zw
cp
Paraat 8 9 10
.
Ioe&ptoe&nij c deen (wie) toeblo'o. s
T:I=MMIIulr!_tt.nenIDm
r'
- .
C S C S C $ C S C $ C S C S C $ C S C S
0
48
.
concer*rade mg
-
tijd
I%Ob
. . .
. . . . . . . . . .
o
_j!;•••
.
.
.
. .
.
.
.°
IMW-R 92/348a
page 31 of 38
RA92348aIMWjt
Table B3 Oxy gen concentration measurements (mg. 1-1).
INS1TTUUT VOOR MIUEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE Fomvijer : IMWB/FW4024 Voor Re9islretie O2-waaien ii verse en gebudde oplossingen in de aaite semt-adie loets met Acartia Ww (48fl); 1 derperproetvat Behoort bij Wedrvoo,sclwdt IMWBISM/4020 .'
Proefcod.: t AT
Concsnt~ mr1 4gJ4 N.L% o
Datum lljds9 Pf tijd (h)
1
-.---
•
Paraat
2 3 4 5 6 7 8 9 10
C S C S C S C S C S C $ C $ C S C $ 0 S
-Iz
0
-
-
24 48
Datum Tiffistip Proeftijd
Conceatratle mgr'4i9r11ve1.% t
Paraat
1 2 3 4 5 6 7 8 9 10 C S C S C $ C $ C S C S C S C S C $ 0 $
24
10
48
-
-
0
c: deai ( schone) loetsoplosskig ile) toetsoplossing
8_.
wfl
zw
0
0
-
-
IMW-R 92/348a
page 32 of 38
RA92348aIMWjt
INSTITUUT VOOB UIUEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE Pmetcode: RT.~ Fr17
Fom'uulier : IMWB/FM4024 Voor Reistretie 02-waarden In verse en gebluide oplossIngen In de aafle semi-statische toets met Aca,tia Ww (481»; 1 Cker per pmetvat BehOOrt b Werkvoo,schrift IMWBFSM/4020
mgI4hgJ4hiol tijd
8 9 10
1 2
C S C $ C $ C $ C $ C $ 0 S C S C $ C S
L!_
°
-
24 48
Datum Tijds9
Con rails mg.r't1NsL% 3z
proef-
- -
1 2 3 4 5 6 7 8 9 10
(t»
C S C S C $ C S C S C S C S C S C $ C S
0 24
---
_:_ 48
- -
c: clean (schone) loetsoptossIng S: speril (vuile) toetsoptossIng
d.nØ -. &IWfl Cum
zw
-
IMW-R 92/348a
page 33 of 38
RA92348aIMWjt
INS1TTUUT VOOR MHJEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE " Fomuber IMWB,1MI4024 voor ReisVie 02-waden In verse en getvWde oplossingen 1 der per poe*vet Behooil bij : Weftvoorsctwilt IMWBISW4020
cbz
----
0
Parf
concwMft fflg tgJ?ve*% 5
T~ Proef-. tijd (*1) -.-------
Pro&code: AT-?
In de aQJte senil-siatisct'ie bets msi Acattia lonsa (48*1);
1 2 3 4 5 6 7 8 9 10 C S C S 0 $ C S C S C S C $ C S C S C S
V,
24
mgX'igri% tijd
8 9 10
1 2
C S C S C S C S C S C S C $ C S C $ 0 S
0
.
24
?. 3
-
- --
.
c clean (schone) betsoplossing S: speri (vuile) toetsopbssing
-. uw Cum
-
-
'7 U
- 0
-
IMW-R 92!348a
page 34 of 38
RA92348aI MWjt
INSTITUUT VOOR MIUEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE Fomuher :I1WB/FM14024 , Voor Registratie 02-waarden In verse en getiniltie oplossingen in de aajte sw*sl 1 ner per — Behoort bij Weoorsctwilt IMWBSW4020
Pmefcode: fçrt
loets met Acartia lonss (48*);
osipnlraRe mgJl/pgJlNot%
tijd (h)
Paraat
1 2 3 4 5 6 7 8 9 10 c S C S C S C S C S C $ C S C S C $ C S
_9
s
0
24
Cøratletng.F1 4kØJ 11v0L%3z
Paraat
tijd (Ii)
1 2 3 4 5 6 7 8 9 10 C $ C S C S C $ C S C S C S C S C S C S
-
o
- -
24
-
1912S.o ø
-
1
clean (Schone) toelsopiosstrug
S: spevi (vuile) loetsoçrbssIng
ewn
-
DuS
Time
5,2 dl.
IMW-R 92/348a
page 35 of 38
RA92348aIMWjt
Table B4 Temperarure measurenients (°C).
INSTITUUT VOOR MILIEUWETENSCHAPPEN TNO AFDEUNG BIOLOGIE Protcod: Forniuher : IMWB/FM/4025 Voor : Registratie van temperatuur in verse en gebnikte oplossingen in de RI acute seni-statische toets met Acartia tonsa (48 h); 1 dier per proetvat Behoort bij : Werkvoorscflritt IMWB/SM14020
Datum Tijdstip Proeftijd (dagen) 0 •
51992 Atdefrig 9olog*, IW.'TNO. Delft 11920501
Temperatuur (°C) s c
____________
Paraat
IMW-R 92/348a
page 36 of 38
RA92348aIMWjt
ANNEX C ESTIMATION OF THE LC50 AND ITS CONFIDENCE INTERVAL
At a given time, the mortality probability of an individual is assessed to be logistically related to the logarithm of the concentration, i.e. pi
- e 1 + P0 where e i = (c / (x)'13 and - 1+ e
p1 is the mortality probability in the ith concentration Po is the mortality probability in concentration o
a is the LC50
f3 is a parameter inversely proportional to the maximum gradient of the dose response function c1 is the ith concentration. The parameters Po' a and 0 are estimated from the counts by means of the maximum likelihood method; i.e. the parameter values to be selected maximize the probability of the counts as a function of the three parameters. Since the distribution of a will not be symmetrical the variance-covariance matrix is not estimated for the parameters Po' (X afld f3 themselves, but for Po' y = in a and 0. The variance-covariance matrix is estimated by the inverse of the information matrix. The 95% confidence limits of the LC50 are now given by exp (± 2 [var (y)] 1/2) = a . exp (± 2 [var (in (X)] 112)
Almo-
zw
IMW-R 92!348a
page 37 of 38
RA92348aIMWjt
ANNEX D CHEMICAL ANALYSIS 1 * * 1
TNO Environmental and Energy Research
TNO Institute of Environmental Scierices Department of Analytical Chemistry Schoemakerstraat 97 P0. Box 6011 2600 JA Delft The Netherlands Telex 38071 zptno ni Fax +31 15 61 68 12 Phone+31 15 69 69 00
QA.UF5ED BYSTERL.5
TNO-report Institute of Envirorunental Sciences TNO Dept. of Biology attn. Drs. R.N. Hooftman P0 Box 6011 2600 JA DELFT
Subj eet: Analysis of praseodyiniuni in water samples from a 48 h test with praseodymium (III) chioride heptahydrate and the marine copepod Acartia tonsa. TNO Study no Report no. Order no. Date Pages
IMW-92-0069-01 TR 92/0595 51808 31-12-92 2
CONTENTS: Statement of GLP Compliance Samples Analytical method and results Quality control Deviations from the protocol All rights reserved. No part of this publication may be reproduced and/or published by pnrtt, photoprint, microfilm or any other means without the previous written consent of TNO.
Issued by: Dept. of Analytical Chemistry E.M. sberg (Technician) Date: '
In oase this report was drafted on instructions, the nghts and obligations of contracting parties are subect to either the Standard Conditions for Research Instructions given to TNO or the relevant agreement conciuded between the contractirtg partres. Submitting the report for inspection to parties who have a direct interest is perrnitted.
rincipal Investigator Date:
© TNO
(Head) Date: Netherlands organuzatlon for applied scientific research Based er the necessity for a sastariabie denelopmerit of society, TNO Environmeptal and Eriergy Research aims at contnbuhtrg, througri research and advice, to adeguate elivironmental management. rationai ertergy COnsumpt.On and the proper management and use of subsurlace natural resources.
4
IlIIt.
T
93
Entered in the 5TERLAB-r'egister for laboratones under number 26 for the areas as defined in the accredtation. liie Standard Corrditions for Research Instructions given to TNO. as fiied at the Registry of the Distnct CouP and the Chamber of Comrnerce in The Hague shali appiy to all instructions given to TNO.
IMW-R 921348a
page 38 of 38
RA92348aIMWjt
1 1
TR 92/ 595
1
page 2
Statement of GLP Compliance The Principal Investigator of the department of Analytical Chemistry identified on the cover page of this report, declares that the work to which this report refers, was carried Out under his supervision according to the procedures herein dèscribed, that to the best of his knowledge this report provides an accurate record of the results obtained and that this part of the study was carried out in compliance with the OECD code of Good L.aboratory Practice. Sanrnles Samples were delivered by the Department of Biology on October 30, 1992. The analyses were performed on November 2, 1992. Analytical method and results The concentration of praseodymium in the solutions was determined by inductively coupled plasma atomic emission spectrometry. The standard solutions were prepared from the test substance. The resuits are listed in the following table. Table 1. Concentration of praseodymium in mg.1' sample code
date of sampling Pr
RIZG AT2 0 1.0 5.6 32
mg.l' mg.1' mg.1 1 mg.1 1
071092 071092 071092 071092
clean clean clean clean
<0.1 <0.1 0.1 2.0
RIZG AT2 0 1.0 5.6 32
mg.1' mg.l' mg.1' mg.1'
091092 091092 091092 091092
spent spent spent spent
<0.1 <0.1 0.2 0.3
Quality control Two samples were also analyzed after the addition of a praseodymium standard solution. The recovery of the additions was 98%. The results are not corrected for the recoveries. The detection limit of 0.1 mg.1' praseodymiuni (see table) is defined as three times the standard deviation of the praseodymium concentration of a blank solution. Deviations from the protocol As from the lst of October, K.J.M. Dejonghe was replaced by M. van Son as Priricipal Investigator. In the protocol should be mentioned that Ms E.M. Stofberg is Technician for the Department of Analytical Chemistry.
OEKO
zw
C23
