SRC TR-11-071

HPV4 CHEMICAL DATA ADEQUACY REVIEW (REVISED)

Plumbane, tetraethyl-, CAS No. 78-00-2

Prepared by:

Carlye Austin, Teresa Manyin, and Joan D. Garey

Chemical, Biological and Environmental Center

SRC, Inc.

7502 Round Pond Road

North Syracuse, NY 13212

Contract No. EP-W-09-027

Task FG007.0.009.001

Submitted to:

OPPT/RAD/HPVCB (7403M)

U.S. Environmental Protection Agency

1200 Pennsylvania Avenue

Washington, D.C. 20460

Debra Milligan, Project Officer

Meena Sonawane, Work Assignment Manager

David Brooks, Alternate Work Assignment Manager

August 5, 2011
HPV4 Chemical Data Adequacy Review (Revised)

Plumbane, tetraethyl-, CAS No. 78-00-2

August 2011

DRAFT

Submitted to:

High Production Volume Chemicals Branch 

Risk Assessment Division 

Office of Pollution Prevention and Toxics 

Environmental Protection Agency 

WAM: Meena Sonawane

Alternate WAM: David Brooks

1200 Pennsylvania Avenue, NW 

Washington, DC 20460-0001

Prepared By:

SRC, Inc. 

Chemical, Biological and Environmental Center

2451 Crystal Drive, Suite 804 

Arlington, VA 22202
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

1

A. PREPARATION OF THE DATA ADEQUACY ASSESSMENTS

A-1. Source material used for assessment

For this unsponsored chemical, a comprehensive literature search was performed to 

identify available toxicity data for the following SIDS endpoints:

Acute toxicity to fish

Acute toxicity to Daphnia

Toxicity to aquatic plants

Acute (oral, dermal, inhalation) toxicity

Repeated-dose toxicity

Reproductive toxicity

Developmental toxicity

Gene mutation

Chromosomal aberrations

Studies relevant to non-SIDS endpoints (e.g., eye and skin irritation; neurotoxicity) conducted 

via oral, inhalation, or dermal routes were also identified through literature searching.

The following sources were searched for toxicity data:

PubMed - Bibliographic database that contains references to journal articles in the life 

sciences with a concentration on biomedicine

TOXNET -National Library of Medicine databases on toxicology, hazardous chemicals, 

environmental health, and toxic releases; includes: 

• TOXLINE with BIOSIS and NTIS;

• Development and Reproductive Toxicology File (DART);

• Chemical Carcinogenesis Research Information System (CCRIS) -

Carcinogenicity and mutagenicity test results for over 8000 

• chemicals);

• Hazardous Substances Data Bank (HSDB) - Comprehensive, peerreviewed toxicology data for about 5000 chemicals;

• Genetic Toxicology Data Bank (GENE-TOX) - Peer-reviewed genetic 

toxicology test data for over 3000 chemicals.

TSCATS - Unpublished technical reports submitted by industry to the EPA under the 

Toxic Substances Control Act]

Registry of toxic effects of chemical substances (RTECs)

Environmental Fate Database (EFDB)

ECOTOX -EPA’s peer-reviewed ecotoxicology database
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

2

EPA’s Integrated Risk Information System (IRIS) website

Agency for Toxic Substances and Disease Registry (ATSDR) website

National Toxicology Program (NTP) website

International Program on Chemical Safety Concise International Chemical Assessment 

Documents (IPCS INCHEM or CICADs)

European Chemical Substances Information System (ESIS) - source of IUCLID Datasets

Any available IUCLID Datasets found through the ESIS website and robust summaries 

submitted to EPA by former sponsors on the assigned chemicals were retrieved. Original 

sources were obtained for any relevant toxicity data found for the assigned chemicals. 

While IUCLID Datasets sometimes contained studies of interest, certain study 

descriptions contained insufficient detail to assess adequacy. In these cases, the primary source 

of the study, if available and accessible, was obtained and reviewed.

A-2. Evaluating data adequacy

Using standard OECD testing guidelines and professional judgment as guides, available 

toxicity studies for the unsponsored chemical were reviewed for adequacy. Summaries were 

prepared for all relevant data found, even if little or no study detail was available. Each study 

was summarized according to a standard format and assigned a Klimisch reliability code; all 

study summaries for the chemical were combined into a single summary document. For studies 

already assigned Klimisch codes (e.g., studies summarized in IUCLID datasets), an explanation 

was provided if the reviewer’s assessment of data adequacy differed from the pre-assigned 

Klimisch code. A table summarizing data availability/adequacy for each SIDS endpoint is 

included at the end of the summary document. 

B. SPECIFIC INFORMATION ON THE AVAILABLE TOXICITY DATA

The available data for each of the SIDS toxicity endpoints are evaluated on the following 

pages. Studies for a common endpoint are grouped together in the document and an endpoint 

adequacy assessment is provided near the top of the first page of the summaries for that 

endpoint. If the data available for the endpoint is determined to be inadequate, a basis for that 

conclusion is supplied. NOTE: If no relevant studies were identified for the chemical, only the 

data availability/adequacy summary table was completed for the report.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

3

B-1. ACUTE TOXICITY TO FISH

Summary of endpoint: Adequate

Study 1: Acute toxicity flow-through study in European plaice

Title: The acute toxicity and bioaccumulation of some lead alkyl compounds in marine animals

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Ellesmere Port Research Laboratories of the Associated Octel 

Company Limited

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish – Flow-through

GLP compliant?: Not specified

Year study was performed: 1980

Test species: Pleuronectes platessa (European plaice; saltwater fish). Mean length: 5.2 cm; 

mean weight: 2.8 g. Source: Tor Bay, S. Devon; obtained via otter trawl.

Analytical monitoring: Yes

Exposure duration: 96 hours

Concentrations tested: 0, 0.02 to 0.2 mg Pb/L

Test method/conditions remarks: All fish were maintained in flowing sea-water for a 

minimum of 7 days prior to test item exposure. Seawater (salinity 34.9‰) was pumped directly 

from the sea and supplied at a constant temperature (15 oC) to the test system. At the design 

flow rate of 100 mL/min, the 90% replacement time was 7.5 hours. 

Stock solutions were prepared by agitating 22 liters of water with 10 grams of tetraethyl lead for 

4 hours, allowing the excess to settle for 2 hours, and then pumping the top 20 liters into a sealed 

stock vessel isolated from direct sunlight. A concentration of 0.02 to 0.2 mg Pb/L tetraethyl lead 

was maintained with a variation of ±10%.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

4

Ten fish were transferred to a test vessel and maintained in flowing seawater for 24 hours, at

which time a pre-selected amount of the stock solution was fed continuously into the vessel for a 

period of 96 hours. A control vessel was included. Test vessels were regularly monitored for 

mortality. Water samples were taken twice daily to analyze tetraethyl lead content. 

Temperature, dissolved oxygen, and pH were measured daily. Test animals were fed prior to but 

not during exposure.

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.23 mg Pb/L.

Results remarks: The LC50 value was calculated according to the methods of Doudoroff et al.

(1951). 

CONCLUSIONS

The study authors reported that the results show that the plaice is more resistant to the effects of 

tetraethyl lead than to tetramethyl lead.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. No guideline or method was cited, GLP 

compliance is unknown, individual data on mortality were not provided, and the exact tested 

concentrations were unclear. Dissolved oxygen, pH, and total organic carbon were not specified. 

Additionally, the test was performed with a non-standard marine species.

REFERENCE

Maddock, B.G. and D. Taylor. 1980. The acute toxicity and bioaccumulation of some lead alkyl 

compounds in marine animals. In: M. Branika and Z. Konrad (eds.), Lead in the Marine 

Environment. Proc. Int. Experts Discussion on Lead Occurrence. Fate and Pollution in the 

Marine Environment, Rovinj, Yugoslavia: 233-261. Pergamon, Oxford, UK.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

5

Study 2: Acute toxicity study in multiple fish species

Title: None specified

This study is: Adequate Not Adequate

Test duration is less than the OECD-recommended length of 96 hours and tests were conducted 

using non-preferred fish species. Additionally, the IUCLID summary, which was mostly in a 

language other than English, lacked study details. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Purity not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish – Flow-through

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1977

Test species: Gobius niger (black goby), Trachurus trachurus (Atlantic horse mackerel), Mugil 

cephalus (striped mullet), Atherina boyeri (sand smelt), Trachinus draco (greater weever)

Analytical monitoring: Not specified

Exposure duration: 30 minutes

Concentrations tested: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): Not specified.

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

6

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. Test duration is less than the OECD-recommended length of 96 

hours and tests were conducted using non-standard fish species. Additionally, the IUCLID 

summary, which was mostly in a language other than English, lacked study details. 

The IUCLID summary noted an assigned Klimisch code of 2 for the study.

REFERENCE

Primary reference (not available): Giaccio, M. 1977. Quad. Merceol. 16: 55-62. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

7

Study 3: Acute toxicity study in bluegill sunfish

Title: Toxicity of various refinery materials to fresh water fish.

This study is: Adequate Not Adequate

Test duration is less than the OECD-recommended length of 96 hours; no analytical monitoring 

was performed; loading rate exceeded the OECD-recommended limit.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Purity not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish – Static

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1954

Test species: Bluegill sunfish (Lepomis macrochirus), obtained from the Pennsylvania State 

Hatchery (Torresdale, PA) and Hiram People’s Hatchery (New Providence, PA). Average 

length: 7 cm. Average weight: 5 g.

Analytical monitoring: No

Exposure duration: 48 hours

Concentrations tested: Not specified

Test method/conditions remarks: The test chemical was added to the 5-gallon test jar at the 

start of the test period. No subsequent additions were made during the course of the test. Test 

jars were filled to the 15-liter mark and placed in a 20 oC thermostatically-controlled water bath. 

Carbon-filtered Philadelphia tap water was used. Fish were acclimatized to the test water and 

temperature for 10 days prior to the test period. Ten fish per jar were tested. Based on the 

average fish weight of 5 g and the number of fish per unit volume of test solution (10 fish/15 L), 

the loading rate was 3.3 g fish/L, which exceeds the OECD-recommended maximum loading 

rate of 1 g for static tests. Characteristics of dilution water: total alkalinity as CaCO3 = 38-81 

ppm; pH = 6.9-7.5; total hardness as CaCO3 = 84-163 ppm; specific conductivity = 228-301 

micromhos; dissolved solids = 154-210 ppm; sulfates as SO4 = 56-98 ppm; chlorides as Cl = 3-

26 ppm. Dissolved oxygen concentration was maintained at 5 ppm or greater throughout the test 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

8

by gentle aeration. 

Mortality was recorded after 24 and 48 hours. The LC50 was obtained by interpolation from a 

graph of the logarithm of the concentration versus the percentage mortality.

RESULTS

Endpoint value(s): The 48-hr LC50 value was 1.4 mg lead/L. The 24-hour LC50 value was 2.0 

mg lead/L.

Results remarks: None.

CONCLUSIONS

No conclusions were specified.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. 

Test duration is less than the OECD-recommended length of 96 hours; no analytical monitoring 

was performed; loading rate exceeded the OECD-recommended limit.

The IUCLID summary noted an assigned Klimisch code of 2 for the study.

REFERENCE

Turnbull, H. et al. 1954. Toxicity of various refinery materials to fresh water fish. Ind. Eng. 

Chem. 46: 324-333.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

9

Study 4: Acute toxicity study in bluegill sunfish

Title: None specified

This study is: Adequate Not Adequate

The IUCLID summary lacked sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Purity not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1964

Test species: Bluegill sunfish (Lepomis macrochirus)

Analytical monitoring: Not specified

Exposure duration: 96 hours

Concentrations tested: Not specified

Test method/conditions remarks: None specified

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.2 mg/L.

Results remarks: None.

CONCLUSIONS

None specified.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

10

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacked sufficient information to 

assess data adequacy.

REFERENCE

Primary reference (not available): Makina, J.F. 1964. Toxicology of petrochemicals in the 

aquatic environment – water and sewage works, vol 3, pp. 456-460. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

11

Study 5: Acute toxicity study in bluegill sunfish

Title: Initial submission: 96-hr LC50 to Bluegill Sunfish 

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Composition of test item (w/w): 61.3% tetraethyl lead; 35.68% 1,2-dibromoethane; 

0.026% blue dye; 2.93% kerosene and inerts; 0.06% antioxidant AO-29.

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish – Static

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1976

Test species: Lepomis macrochirus (Bluegill sunfish); 3.6 cm standard length.

Analytical monitoring: Not indicated

Exposure duration: 96 hours

Concentrations tested: 0, 0.010, 0.025, 0.050, 0.075, 0.10, 0.25, 0.5 mg/L

Test method/conditions remarks: The test material, as either a 0.1 or 0.01% solution in 

acetone, was introduced into all-glass vessels containing ten bluefish in 15 liters of water. Fish 

were not fed for 48 hours prior to testing, nor during exposure. No aeration was provided during 

the test. Dissolved oxygen and pH were measured at the beginning of exposure. Calculation of 

the 96-hour LC50 was via straight line graphical interpolation. An acetone control was included, 

and prepared by adding, to clean supply water, a volume of acetone equal to the greatest volume 

of test solution used. A negative water control was also included.

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.15 mg/L.

Results remarks: The test temperature was maintained at 20 ± 0.2 oC. The pH measurements 

across tested concentrations ranged from 7.7 to 8.0 and dissolved oxygen ranged from 8.6 to 8.7 

ppm. Mortality was 100% at 0.25 and 0.50 mg/L, 10% at 0.10 mg/L, and 0% at all other tested 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

12

concentrations.

CONCLUSIONS

Tetraethyl lead exhibits extreme acute toxicity to bluegill sunfish.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. No guideline or method was cited, and GLP 

compliance is unknown. Additionally, it is unclear whether analytical monitoring was performed

and total organic carbon was not specified.

REFERENCE

TSCATS submission (OTS0546521, Doc ID 88-920010136, received 10/27/92): Haskell 

Laboratory, DuPont Chemical Company. 1976. Initial submission: 96-hr LC50 to Bluegill 

Sunfish with Cover Letter Dated 101592.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

13

Study 6: Acute toxicity study in fathead minnows

Title: Initial submission: Report of 96-hr LC50 of Lead, Tetraethyl in Fathead Minnows 

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Composition of test item (w/w): 61.3% tetraethyl lead; 35.68% 1,2-dibromoethane; 

0.026% blue dye; 2.93% kerosene and inerts; 0.06% antioxidant AO-29.

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish – Static

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1975

Test species: Pimephales promelas (Fathead minnow); 3.7 cm standard length.

Analytical monitoring: Not indicated

Exposure duration: 96 hours

Concentrations tested: 0, 0.005, 0.0075, 0.01, 0.025, 0.05, 0.075, or 0.1 mg/L

Test method/conditions remarks: The test material, as a 10, 0.1, or 0.01% solution in acetone, 

was introduced into all-glass vessels containing ten fathead minnows in 15 liters of water. Fish 

were not fed during exposure. No aeration was provided during the test, but test vessels were 

vigorously aerated for 30 minutes prior to introduction of fish and test material. The pH was 

measured at the beginning of exposure. Calculation of the 96-hour LC50 was via straight line 

graphical interpolation. An acetone control was included, and prepared by adding, to clean 

supply water, a volume of acetone equal to the greatest volume of test solution used. A negative

water control was also included.

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.037 mg/L.

Results remarks: The test temperature was maintained at 20 ± 0.2 oC. The pH was 7.8. 

Mortality was 100% at the two highest test concentrations, 90% at the third-highest 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

14

concentration, and 0% at all other tested concentrations.

CONCLUSIONS

Tetraethyl lead exhibits extreme acute toxicity to fathead minnows.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. No guideline or method was cited, and GLP 

compliance is unknown. Additionally, it is unclear whether analytical monitoring was 

performed, and dissolved oxygen concentrations and total organic carbon were not measured.

REFERENCE

TSCATS submission (OTS0555568, Doc ID 88-920010086, received 10/27/92): Haskell 

Laboratory, DuPont Chemical Company. 1975. Initial submission: Report of 96-hr LC50 of 

Lead, Tetraethyl in Fathead Minnows with Cover Letter Dated 101592.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

15

Study 7: Acute toxicity study in European seabass larvae

Title: Acute toxicity of alkyl leads to some marine organisms

This study is: Adequate Not Adequate

The test duration was less than the OECD-recommended length of 96 hours. Additionally, the 

test substance was a formulation (composition not specified); the tested concentrations were 

unclear; a non-standard fish species was tested; and missing experimental details included 

loading rate, pH, salinity, dissolved oxygen concentration, total organic carbon levels, and 

whether the test was conducted in static or flow-through chambers.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Purity not provided; test substance described as a commercial product, “TEL-CB”

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Fish

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1978

Test species: 6-mm larvae of Morone labrax (European seabass)

Analytical monitoring: No

Exposure duration: 48 hours

Concentrations tested: Not specified

Test method/conditions remarks: The tetraethyl lead solution was prepared by adding 2 ml of 

product to 1 L of filtered natural seawater (0.45 μm) and slowly stirring magnetically for 1 hour. 

The upper one fourth of the saturated solution was used for preparing experimental dilutions in 

natural filtered seawater. Analytical measurement by atomic absorption spectrophotometry gave 

a content of 2 mg total lead/L for the stock solution. The experiment was carried out in the dark, 

at 20 oC.

RESULTS

Endpoint value(s): The 48-hr LC50 was 65 ppb Pb. One-hundred percent mortality was seen 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

16

after 48 hours at 130 ppb Pb.

Results remarks: All larvae died almost immediately at concentrations of 650 ppb Pb. There 

were no mortalities at 10 ppb Pb.

CONCLUSIONS

Alkyl leads show higher toxicity than inorganic lead.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The test duration was less than the OECD-recommended length of 

96 hours. Additionally, the tested concentrations were unclear; a non-standard fish species was 

tested; and missing experimental details included loading rate, pH, salinity, dissolved oxygen 

concentration, total organic carbon levels, and whether the test was conducted in static or flowthrough chambers.

REFERENCE

Marchetti, R. 1978. Acute toxicity of alkyl leads to some marine organisms. Marine Pollution 

Bulletin, 9: 206-207.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

17

B-2. ACUTE TOXICITY TO DAPHNIA

Summary of endpoint: Adequate

Study 1: Acute toxicity flow-through study in brown shrimp

Title: The acute toxicity and bioaccumulation of some lead alkyl compounds in marine animals

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 9

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Aquatic Invertebrates – Flow-through

GLP compliant?: Not specified

Year study was performed: 1980

Test species: Crangon crangon (brown shrimp). Mean length: 4.8 cm; mean weight: 1.1 g. 

Source: Tor Bay, S. Devon; obtained via push net.

Analytical monitoring: Yes

Exposure duration: 96 hours

Concentrations tested: 0.01 to 0.2 mg Pb/L

Test method/conditions remarks: All shrimp were maintained in flowing sea-water for a 

minimum of 7 days prior to test item exposure. Seawater (salinity 34.9‰) was pumped directly 

from the sea and supplied at a constant temperature (15 oC) to the test system. At the design 

flow rate of 100 mL/min, the 90% replacement time was 7.5 hours. 

Stock solutions were prepared by agitating 22 liters of water with 10 grams of tetraethyl lead for 

4 hours, allowing the excess to settle for 2 hours, and then pumping the top 20 liters into a sealed 

stock vessel isolated from direct sunlight. A concentration of 0.01 to 0.2 mg Pb/L tetraethyl lead 

was maintained with a variation of ±10%.

Ten shrimp were transferred to a test vessel and maintained in flowing seawater for 24 hours, at 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

18

which time pre-selected amount of the stock solution was fed continuously into the vessel for a 

period of 96 hours. A control vessel was included. Test vessels were regularly monitored for 

mortality. Water samples were taken twice daily to analyze tetraethyl lead content. 

Temperature, dissolved oxygen, and pH were measured daily. Test animals were fed prior to but 

not during exposure.

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.02 mg Pb/L.

Results remarks: The LC50 value was calculated according to the methods of Doudoroff et al.

(1951). 

CONCLUSIONS

None specified.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. No guideline or method was cited, GLP 

compliance is unknown, individual data on mortality were not provided, and the exact tested 

concentrations were unclear. Additionally, the test employed a non-standard species and 

dissolved oxygen, pH, and total organic carbon were not specified.

REFERENCE

Maddock, B.G. and D. Taylor. 1980. The acute toxicity and bioaccumulation of some lead alkyl 

compounds in marine animals. In: M. Branika and Z. Konrad (eds.), Lead in the Marine 

Environment. Proc. Int. Experts Discussion on Lead Occurrence. Fate and Pollution in the 

Marine Environment, Rovinj, Yugoslavia: 233-261. Pergamon, Oxford, UK.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

19

Study 2: Acute toxicity flow-through study in blue mussel

Title: The acute toxicity and bioaccumulation of some lead alkyl compounds in marine animals

This study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Ellesmere Port Research Laboratories of the Associated Octel 

Company Limited

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Aquatic Invertebrates – Flow-through

GLP compliant?: Not specified

Year study was performed: 1980

Test species: Mytilus edulis (blue mussel). Mean length: 6.4 cm; mean weight: 28.5 g. Source: 

Teign Estuary, S. Devon; obtained via hand collection.

Analytical monitoring: Yes

Exposure duration: 96 hours

Concentrations tested: 0.01 to 0.2 mg Pb/L

Test method/conditions remarks: All mussels were maintained in flowing sea-water for a 

minimum of 7 days prior to test item exposure. Seawater (salinity 34.9‰) was pumped directly 

from the sea and supplied at a constant temperature (15 oC) to the test system. At the design 

flow rate of 100 mL/min, the 90% replacement time was 7.5 hours. 

Stock solutions were prepared by agitating 22 liters of water with 10 grams of tetraethyl lead for 

4 hours, allowing the excess to settle for 2 hours, and then pumping the top 20 liters into a sealed 

stock vessel isolated from direct sunlight. A concentration of 0.01 to 0.2 mg Pb/L tetraethyl lead 

was maintained with a variation of ±10%.

Ten mussels were transferred to a test vessel and maintained in flowing seawater for 24 hours, at 

which time a pre-selected amount of the stock solution was fed continuously into the vessel for a 

period of 96 hours. A control vessel was included. Test vessels were regularly monitored for 

mortality. Water samples were taken twice daily to analyze tetraethyl lead content. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

20

Temperature, dissolved oxygen, and pH were measured daily. Test animals were fed prior to but 

not during exposure.

RESULTS

Endpoint value(s): The 96-hour LC50 value was 0.10 mg Pb/L.

Results remarks: The LC50 value was calculated according to the methods of Doudoroff et al.

(1951). 

CONCLUSIONS

None specified.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. No guideline or method was cited, GLP 

compliance is unknown, individual data on mortality were not provided, and the exact tested 

concentrations were unclear. Additionally, the test was conducted with a non-standard species 

and dissolved oxygen, pH, and total organic carbon were not specified.

REFERENCE

Maddock, B.G. and D. Taylor. 1980. The acute toxicity and bioaccumulation of some lead alkyl 

compounds in marine animals. In: M. Branika and Z. Konrad (eds.), Lead in the Marine 

Environment. Proc. Int. Experts Discussion on Lead Occurrence. Fate and Pollution in the 

Marine Environment, Rovinj, Yugoslavia: 233-261. Pergamon, Oxford, UK.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

21

Study 3: Acute toxicity study in Eastern Oyster

Title: Bioaccumulation and Depuration of Lead by Eastern Oysters (Crassostrea virginica) 

Exposed to a Tetraethyl Lead Motor Fuel Antiknock Compound

This study is: Adequate Not Adequate

The concentration of tetraethyl lead in the test substance was not specified.

TEST SUBSTANCE 

Identity: TEL Motor Antiknock-Red-01, a tetraethyl lead motor fuel antiknock compound

Remarks: Concentration of tetraethyl lead not specified; lead content: 0.6247 g Pb/mL; clear red 

liquid; solvent/carrier: reagent grade absolute ethanol

METHOD

Method/guideline followed: Methods of Butler et al. (1960) and Butler (1965)

Test type: Acute Toxicity to Aquatic Invertebrates – shell deposition test (flow-through)

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1976

Test species: Eastern oyster (Crassostrea virginica), 2-4 cm from umbo to distal valve edge, 

wild-caught (Pensacola, FL)

Analytical monitoring: Yes

Exposure duration: 96 hours

Concentrations tested: Nominal: 0 (solvent control), 1.0, 10, 56, 100, 200 μg Pb/L; measured: 

0, 1.0, 5.2, 36, 72, 131 μg Pb/L

Test method/conditions remarks: Oysters were held in tanks of flowing, natural seawater of 

ambient salinity and temperature for a minimum of two weeks before testing. Oysters were 

cleaned of attached organisms and ground by hand on a fine-grit grinding wheel to remove 

approximately 2-5 mm of peripheral shell. Six groups of ten oysters each were exposed to each 

test concentration for 96 hours. The test was conducted in a glass system that was covered, 

where feasible, to reduce volatilization of the test substance. The system was 18 L, received 

seawater at a rate of 30 L/hour, and ~99% replacement occurred every 3 hours. New shell 

growth was measured to the nearest 0.1 mm using a binocular dissecting scope equipped with a 

micrometer.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

22

RESULTS

Endpoint value(s): The 96-hour EC50 value for shell growth was > 1 but < 5.2 μg Pb/L.

Results remarks: The percentage reduction of new shell growth of exposed oysters, as 

compared to control oysters was 43, 85, 85, 85, and 100% at concentrations of 1.0, 5.2, 36, 72, 

and 131 μg Pb/L, respectively. Salinity was 30-32‰ and temperature was 28-30 °C.

CONCLUSIONS

Shell growth of oysters exposed to the test substance in seawater was reduced by more than 50% 

in concentrations of tetraalkyl lead ≥ 5.2 μg Pb/L during an acute exposure.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The concentration of tetraethyl lead in the test substance was not 

specified.

REFERENCE

TSCATS submission (OTS0200597, Doc ID 88-7800249, received 12/10/79): Haskell 

Laboratory, EI DuPont de Nemours & Company. 1978. Bioaccumulation and Depuration of 

Lead by Eastern Oysters (Crassostrea virginica) Exposed to a Tetraethyl Lead Motor Fuel 

Antiknock Compound with Attachments and Cover Letter Dated 120379, conducted by EG&G 

Bionomics, Marine Research Laboratory, Appendix A. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

23

Study 4: Acute toxicity study in brine shrimp

Title: Acute toxicity of alkyl leads to some marine organisms

This study is: Adequate Not Adequate

The test substance was a formulation; composition was not specified. Additionally, the tested 

concentrations were unclear; a non-standard species was tested; and missing experimental details 

included loading rate, pH, salinity, dissolved oxygen concentration, total organic carbon levels, 

and whether the test was conducted in static or flow-through chambers.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: None provided

METHOD

Method/guideline followed: Not specified

Test type: Acute Toxicity to Aquatic Invertebrates 

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1978

Test species: 24-hour-old nauplii of Artemia salina (brine shrimp)

Analytical monitoring: No

Exposure duration: 48 hours

Concentrations tested: Not specified

Test method/conditions remarks: The tetraethyl lead solution was prepared by adding 2 ml of 

product to 1 L of filtered natural seawater (0.45 μm) and slowly stirring magnetically for 1 hour. 

The upper one fourth of the saturated solution was used for preparing experimental dilutions in 

natural filtered seawater. Analytical measurement by atomic absorption spectrophotometry gave 

a content of 2 mg total lead/L in the stock solution. The experiment was carried out in the dark, 

at 20 oC.

RESULTS

Endpoint value(s): The 48-hr LC50 was 85 ppb Pb. One-hundred percent mortality was seen 

after 48 hours at 260 ppb Pb.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

24

Results remarks: Almost immediate 100% mortality occurred at 770 ppb Pb. No effect was 

seen after 6 hours at 500 ppb Pb.

CONCLUSIONS

Alkyl leads show higher toxicity than inorganic lead.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The test substance was a formulation; composition was not 

specified. Additionally, the tested concentrations were unclear; a non-standard species was 

tested; and missing experimental details included loading rate, pH, salinity, dissolved oxygen 

concentration, total organic carbon levels, and whether the test was conducted in static or flowthrough chambers.

REFERENCE

Marchetti, R. 1978. Acute toxicity of alkyl leads to some marine organisms. Marine Pollution 

Bulletin, 9: 206-207.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

25

B-3. TOXICITY TO AQUATIC PLANTS

Summary of endpoint: Inadequate

Study 1: Acute toxicity study in marine diatom

Title: The acute toxicity and bioaccumulation of some lead alkyl compounds in marine animals

This study is: Adequate Not Adequate

The exposure period appears to have been only 6 hours in duration; OECD guidelines specify a 

duration of 72 hours. Additionally, no guideline or method was cited, GLP compliance is 

unknown, a non-standard test method was used, and the exact tested concentrations were unclear. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Ellesmere Port Research Laboratories of the Associated Octel 

Company Limited

METHOD

Method/guideline followed: Based on a technique developed from the organic productivity 

assessment method of Steeman-Nielsen (1952).

Test type: Toxicity to Algae – Static

GLP compliant?: Not specified

Year study was performed: 1980

Test species: Phaeodactylum tricornutum (marine diatom). Source: Plymouth collection; subculture on modified Erd-Schrieber.

Analytical monitoring: Yes

Exposure duration: 6 hours

Concentrations tested: Not specified

Test method/conditions remarks:

Seawater (salinity 34.9‰) was pumped directly from the sea and supplied at a constant 

temperature (15 oC) to the test system. Stock solutions were prepared by agitating 22 liters of 

water with 10 grams of tetraethyl lead for 4 hours, allowing the excess to settle for 2 hours, and 

then pumping the top 20 liters into a sealed stock vessel isolated from direct sunlight.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

26

A standardized quantity of algae was incubated for 6 hours in the presence of a range of 

concentrations of the test item and a standard amount of C14-labeled carbonate. The 

accumulation of C14 by the control cultures indicates the “normal” activity of the algae which 

can be compared to activity of algae exposed to the test item.

RESULTS

Endpoint value(s): The 6-hour EC50 value was 0.10 mg Pb/L, based on a reduction in 

photosynthetic activity.

Results remarks: The EC50 value was calculated according to the methods of Doudoroff et al.

(1951)

CONCLUSIONS

None specified.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The exposure period appears to have been only 6 hours in 

duration; OECD guidelines specify a duration of 72 hours. Additionally, no guideline or method 

was cited, GLP compliance is unknown, a non-standard test method was used, and the exact 

tested concentrations were unclear. 

REFERENCE

Maddock, B.G. and D. Taylor. 1980. The acute toxicity and bioaccumulation of some lead alkyl 

compounds in marine animals. In: M. Branika and Z. Konrad (eds.), Lead in the Marine 

Environment. Proc. Int. Experts Discussion on Lead Occurrence. Fate and Pollution in the 

Marine Environment, Rovinj, Yugoslavia: 233-261. Pergamon, Oxford, UK.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

27

Study 2: Toxicity study in diatom

Title: Not specified

This study is: Adequate Not Adequate

The IUCLID summary, which was mostly in a language other than English, contains insufficient 

information to assess data adequacy.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Toxicity to Algae

GLP compliant?: Not specified

Year study was performed: 1992

Test species: Cyclotella meneghiniana f. unipunctata (diatom)

Analytical monitoring: Not specified

Exposure duration: 24 hours

Concentrations tested: Not specified

Test method/conditions remarks: Growth rate was measured.

RESULTS

Endpoint value(s): Not specified

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

28

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which was mostly in a language 

other than English, contained insufficient details to assess data adequacy.

The IUCLID dataset summary reported an assigned Klimisch code of 2.

REFERENCE

Primary reference (unavailable): Giri, B.S. and N.A. Devi. 1992. Indian J Exp Biol 30: 201-

204. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

29

Study 3: Toxicity study in green algae

Title: Acute toxicity of alkyl leads to some marine organisms

This study is: Adequate Not Adequate

The test duration was less than the OECD-recommended length of 72 hours. Additionally, the 

test substance was a formulation (composition not specified); the tested concentrations were 

unclear; a non-standard species was tested; and missing experimental details included pH, 

salinity, and dissolved oxygen concentration.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: None provided

METHOD

Method/guideline followed: Not specified

Test type: Toxicity to Algae

GLP compliant?: The study pre-dates GLP.

Year study was performed: 1978

Test species: Dunaliella tertiolecta (green algae)

Analytical monitoring: No

Exposure duration: 48 hours

Concentrations tested: Not specified

Test method/conditions remarks: The tetraethyl lead solution was prepared by adding 2 ml of 

product to 1 L of filtered natural seawater (0.45 μm) and slowly stirring magnetically for 1 hour. 

The upper one fourth of the saturated solution was used for preparing experimental dilutions in 

natural filtered seawater. Analytical measurement by atomic absorption spectrophotometry gave 

a content of 2 mg total lead/L in the stock solution. The experiment was carried out at 20 oC 

using closed vessels to avoid gas-phase leaching of the volatile compound. The toxicity to algae 

was measured as the reduction of photosynthetic activity by the 14C method.

RESULTS

Endpoint value(s): The 48-hr EC50 was 150 ppb. One-hundred percent reduction of 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

30

photosynthesis was seen after 48 hours at 300 ppb.

Results remarks: No effects on photosynthesis were observed at concentrations less than 100 

ppb Pb. Complete inhibition of photosynthetic activity was observed at 500 ppb after 6 hours.

CONCLUSIONS

Alkyl leads show higher toxicity than inorganic lead.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The test duration was less than the OECD-recommended length of 

72 hours. Additionally, the test substance was a formulation (composition not specified); the 

tested concentrations were unclear; a non-standard species was tested; and missing experimental 

details included pH, salinity, and dissolved oxygen concentration.

REFERENCE

Marchetti, R. 1978. Acute toxicity of alkyl leads to some marine organisms. Marine Pollution 

Bulletin, 9: 206-207.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

31

B-4. ACUTE TOXICITY

Study 1: Acute oral toxicity study in rats

Summary of endpoint: Adequate

Title: The Effects of Fasting on the Acute Oral Toxicity of Nine Chemicals in the Rat

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2, supplied by E.I. du Pont de Nemours

Remarks: Purity 98.5%, containing 1% ethyl chloride. Tested as a 0.10-0.20% suspension.

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: Not specified

Year study performed: 1984

Test species: Male Crl-CD rats (Charles River Breeding Laboratory, North Wilmington, MA), 

60 days old, weighing 220-280 grams

Number of animals/sex/dose: 10 males/dose level

Route of administration: Oral

Vehicle: Corn oil

Doses/concentrations tested: 15, 20, 22, 26, 30, 35, or 40 mg/kg for fasted rats; 17, 22, or 28 

mg/kg for non-fasted rats

Statistical methods used: LD50 values were estimated by probit analysis.

Test method/conditions remarks: Rats received a single oral gavage dose of the test substance. 

Seven groups of 24-hour fasted rats and three groups of non-fasted rats were tested. A postdosing 14-day observation period was included, during which rats were weighed and observed 

daily.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

32

RESULTS

Endpoint value(s): The oral LD50 value was 29 mg/kg for fasted rats and 20 mg/kg for nonfasted rats.

Results remarks: Mortality (out of 10 rats at each concentration) was 0, 0, 3, 7, 6, 4, and 9 at 

15, 20, 22, 26, 30, 35, and 40 mg/kg, respectively, for fasted rats and 3, 6, and 10 at 17, 22, and 

28 mg/kg, respectively, for non-fasted rats. At 22 mg/kg (the only dose level common to both 

fasted and non-fasted groups), the number of deaths among non-fasted rats was greater. Time of 

death in both fasted and non-fasted groups was similar, with delayed deaths occurring in both 

groups. The mean time to death was greater in the low-dose groups. Clinical signs of toxicity 

included tremors, convulsions, weakness, diarrhea, and weight loss. Surviving fasted rats 

showed only an immediate transient weight loss, whereas non-fasted rats had a mild to moderate 

decrease followed by recovery two to three days post-dosing.

CONCLUSIONS

Tetraethyl lead would be considered extremely toxic in both fasted and non-fasted rats. LD50

values in fasted and non-fasted rats were not statistically different.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. The study appears generally acceptable, 

although it is missing information including guideline followed and GLP compliance, and only 

male rats were tested.

REFERENCE

Dashiell, O.L. and G.L. Kennedy. 1984. The effects of fasting on the acute oral toxicity of nine 

chemicals in the rat. J. Appl. Toxicol., 4: 320-325. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

33

Study 2: Acute oral toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary lacked sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1972

Test species: Rat

Number of animals/sex/dose: Not specified

Route of administration: Oral

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: Not specified

RESULTS

Endpoint value(s): The oral LD50 value was 12.3 mg/kg.

Results remarks: Not specified
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

34

CONCLUSIONS

Not specified

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacked sufficient information to 

assess data adequacy. 

REFERENCE

Primary reference (unavailable): Experientia, vol 28, p. 923. 1972. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

35

Study 3: Acute oral toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary, which was mostly in a language other than English, lacked sufficient 

information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1963

Test species: Rat

Number of animals/sex/dose: Not specified

Route of administration: Oral

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): The oral LD50 value was 35 mg/kg.

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

36

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which was mostly in a language 

other than English, lacked sufficient information to assess data adequacy. 

The IUCLID summary reported an assigned Klimisch code of 2.

REFERENCE

Primary reference (unavailable): Magistretti, M. et al. 1963. Med Lavoro 54: 486-495. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

37

Study 4: Acute oral toxicity study in rats

Title: The LD50 value of tetraethyl lead

The study is: Adequate Not Adequate

The study tested an insufficient number of animals per dose level. Additionally, information was 

missing on guideline followed and mortality by sex. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure solution, obtained from Ventor Corporation, Beverly, MA. Dissolved in 

100% pure peanut oil.

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1972

Test species: Sprague-Dawley albino rats, weighing from 150 to 250 g (females) and 350 to 425 

g (males)

Number of animals/sex/dose: 2/sex/group

Route of administration: Oral

Vehicle: Peanut oil

Doses/concentrations tested: The test item was dissolved in peanut oil to obtain a concentration 

of 7.32 mg/mL. Pure peanut oil was added as necessary to bring the volume administered up to 

2 mL. Dose levels were 10, 13, 16.9, or 21.97 mg/kg.

Statistical methods used: Weil’s method of moving averages was used.

Test method/conditions remarks: Animals received a single oral gavage dose. All animals 

were kept in the same 12-hr-light-cycled, temperature-controlled room. Animals were examined 

daily for 14 days post-dosing. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

38

RESULTS

Endpoint value(s): The oral LD50 value was 14.18 mg/kg.

Results remarks: Mortalities were as follows: 1/4 at 10 mg/kg, 0/4 at 13 mg/kg, 4/4 at 16.9 

mg/kg, and 4/4 at 21.97 mg/kg. All animals displayed neurological signs including lethargy, 

irritability, and ataxia. Violent jumping, trembling, and thrashing following a loud noise, plus 

aggressiveness and fighting, were also observed. Aggressiveness was seen mostly in the 16.9 

and 21.97 mg/kg groups beginning on day 5 post-dosing. This was followed by convulsions, 

intermittent uncontrolled thrashing, and finally death beginning on day 6 post-dosing.

CONCLUSIONS

None specified

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The study tested an insufficient number of animals per dose level. 

Additionally, information was missing on guideline followed and mortality by sex. 

The IUCLID summary reported an assigned Klimisch code of 2.

REFERENCE

Schroeder, T. et al. 1972. The LD50 value of tetraethyl lead. Experientia, 28: 425-426.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

39

Study 5: Acute oral toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary lacked sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1973

Test species: Rat

Number of animals/sex/dose: Not specified

Route of administration: Oral

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: Not specified

RESULTS

Endpoint value(s): The oral LDLO value was 30 mg/kg.

Results remarks: Not specified

CONCLUSIONS

Not specified
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

40

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacked sufficient information to 

assess data adequacy. 

REFERENCE

Primary reference (unavailable): Japanese Journal of Industrial Health, 15: p. 3. 1973. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

41

Study 6: Acute oral toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary, which was mostly in a language other than English, lacked sufficient 

information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1974

Test species: Rat

Number of animals/sex/dose: Not specified

Route of administration: Oral

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): None specified

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

42

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which was mostly in a language 

other than English, lacked sufficient information to assess data adequacy. 

The IUCLID summary indicated an assigned Klimisch code of 2.

REFERENCE

Primary reference (unavailable): Avery D.D. et al. 1974. Pharmacol. Biochem. Behav. 2: 473-

479. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

43

Study 7: Acute oral toxicity study in rats

Title: Tetraethyllead and tetramethyllead

The study is: Adequate Not Adequate

No LD50 value was specified in the report. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Dissolved in peanut oil to yield a concentration of 1.813 mg/mL; toluene added (as a 

stabilizer) at 0.321 mL per 100 mL peanut oil

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1964

Test species: Charles River cesarean-derived albino rats (22 days old)

Number of animals/sex/dose: 6/sex/dose

Route of administration: Oral

Vehicle: Peanut oil/toluene

Doses/concentrations tested: 0, 1.7, or 17 mg/kg

Statistical methods used: Not specified

Test method/conditions remarks: Animals were housed in stainless steel wire cages and 

observed for 9 days pre-dosing. Pre-dosing hematological data were obtained. Four male and 

for female rats were sacrificed for histologic examination pre-dosing. 

Animals received single oral gavage doses of the test item, were observed for 144 days postdosing, and then sacrificed. Terminal measurements included body weight, organ weights, lead 

content in tissues, and histopathology.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

44

RESULTS

Endpoint value(s): None specified

Results remarks: At 17 mg/kg, 4/6 males and 1/6 females died within 5 days of dosing. All rats 

displayed irritability, hypermotility, tremors, spasticity, and arched backs, and some became 

prostrate. All lost weight within the first week. Within 2 weeks of administration, the surviving

rats appeared normal. At 1.7 mg/kg, there were no deaths or abnormalities observed. No 

adverse effects on terminal body weight were observed. 

Some slight organ weight changes were seen with respect to controls, but no statistical analysis 

was provided. Pathological changes included hypoplasia and hypotrophy of hepatocytes, neural 

degeneration, and hypertrophy of myocardial fibers. The liver was found to contain the greatest 

lead content, relative to other organs.

CONCLUSIONS

Not specified

STUDY RELIABILITY

Klimisch code = [3]. Invalid. No LD50 value was specified in the report. 

The IUCLID summary indicated an assigned Klimisch code of 2.

REFERENCE

Schepers, G.W.H. 1964. Tetraethyllead and tetramethyllead. Arch. Environ. Health, 8: 277-

295.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

45

Study 8: Acute oral toxicity study in rats

Title: Acute Oral Toxicity Study of Tetraethyllead

The study is: Adequate Not Adequate

Insufficient information was available to assess data adequacy; details missing included test 

substance purity, doses tested, test conditions, and mortality at each dose. Additionally, only one 

sex of rats was tested.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Described as Haskell #9380 (TEL MM-I)

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1975

Test species: Young adult male ChR-CD rats

Number of animals/sex/dose: 10/dose

Route of administration: Oral

Vehicle: Corn oil

Doses/concentrations tested: 5 or 50 mg/kg

Statistical methods used: Not specified

Test method/conditions remarks: Animals received single oral gavage doses of the test 

substance, and were then observed for 14 days.

RESULTS

Endpoint value(s): The acute oral LD50 value was >5 but <50 mg/kg.

Results remarks: All rats died at 50 mg/kg. No mortality was observed at 5 mg/kg. At 50 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

46

mg/kg, piloerection and tremors were seen two days after dosing, as well as weight loss, and 

finally death 2 to 3 days post-dosing. At 5 mg/kg, initial weight loss was observed. 

CONCLUSIONS

Tetraethyl lead was classified as highly toxic.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. Insufficient information was available to assess data 

adequacy; details missing included test substance purity, doses tested, test conditions, and 

mortality at each dose. Additionally, only one sex of rats was tested.

REFERENCE

TSCATS submission (OTS0571634, Document ID 88-920009976, received 10/15/92). Haskell 

Laboratory, DuPont Chemical Company. 1975. Special Oral Test of Lead, Tetraethyl in Rats 

with Cover Letter Dated 101592. 

 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

47

Study 9: Acute oral toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary, which was mostly in a language other than English, lacked sufficient 

information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1976

Test species: Rat

Number of animals/sex/dose: Not specified

Route of administration: Oral

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): None specified

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

48

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which was mostly in a language 

other than English, lacked sufficient information to assess data adequacy. 

The IUCLID summary reported an assigned Klimisch code of 2.

REFERENCE

Primary reference (unavailable): Czech et al. 1976. Pharmacol Biochem Behav 5: 387-389. 

Secondary reference: In IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

49

Study 10: Acute oral toxicity study in rats

Title: Teratogenic evaluation of lead compounds in mice and rats

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Houston Chemical Company, Freeport, TX

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1975

Test species: COBS random-bred albino rats

Number of animals/sex/dose: 10/dose, sex not specified

Route of administration: Oral gavage

Vehicle: Not specified

Doses/concentrations tested: 35.12, 52.67, and 79.01 mg/kg

Statistical methods used: The methods of Litchfield and Wilcoxon (1949) were used.

Test method/conditions remarks: Single graded oral doses were given to groups of 10 animals 

and 14-day LD50 values were calculated.

RESULTS

Endpoint value(s): LD50 = 53 ± 8 mg/kg

Results remarks: Mortalities were 1/10, 5/10, and 10/10 at 35.12, 52.67, and 79.01 mg/kg 

respectively. Clinical signs at 52.67 mg/kg included hypoactivity from 6 to 24 hours. Clinical 

signs seen at 79.01 mg/kg included hypoactivity (from 6 hours until death), tremors, from 5 to 24 

hours, or until death), and clonic convulsions (from 2 to 5 hours or until death).
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

50

CONCLUSIONS

None specified

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. The sex of test animals was not specified.

REFERENCE

Kennedy, G.L. et al. 1975. Teratogenic evaluation of lead compounds in mice and rats. Food. 

Cosmet. Toxicol., 13: 629-632.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

51

Study 11: Acute oral toxicity study in mice

Title: Teratogenic evaluation of lead compounds in mice and rats

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Houston Chemical Company, Freeport, TX

METHOD

Method/guideline followed: Not specified

Test type: Acute oral

GLP compliant?: The study pre-dates GLP.

Year study performed: 1975

Test species: CD-1 random-bred albino mice

Number of animals/sex/dose: 10/dose, sex not specified

Route of administration: Oral gavage

Vehicle: Not specified

Doses/concentrations tested: 35.12, 52.67, 79.01, and 118.5 mg/kg

Statistical methods used: The methods of Litchfield and Wilcoxon (1949) were used.

Test method/conditions remarks: Single graded oral doses were given to groups of 10 animals 

and 14-day LD50 values were calculated.

RESULTS

Endpoint value(s): LD50 = 65 ± 3.5 mg/kg

Results remarks: Mortalities were 0/10, 2/10, 10/10, and 10/10 at 35.12, 52.67, 79.01, and 

118.5 mg/kg, respectively. Clinical signs at 52.67 mg/kg included hypoactivity from 6 to 24 

hours. Clinical signs seen at 79.01 mg/kg included hypoactivity (from 6 hours or 2 hours until 

death) and tremors (from 5 to 24 hours, or until death). Clinical signs seen at 118.5 mg/kg 

consisted of hypoactivity (from 2 hours until death), tremors (from 3 to 12 hours or until death) 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

52

and clonic convulsions (from 2-3 to 5 hours).

CONCLUSIONS

None specified

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. The sex of test animals was not specified.

REFERENCE

Kennedy, G.L. et al. 1975. Teratogenic evaluation of lead compounds in mice and rats. Food. 

Cosmet. Toxicol., 13: 629-632.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

53

Study 12: Acute oral toxicity study in rats

Title: Estimation of acute oral toxicity in rats by determination of the approximate lethal dose 

rather than the LD50

The study is: Adequate Not Adequate

Only one rat per dose level was tested.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98.5% pure, obtained from EI du Pont de Nemours and Co., Inc.

METHOD

Method/guideline followed: Not specified

Test type: Acute oral 

GLP compliant?: Not specified

Year study performed: 1986

Test species: Crl:CD male rats, from Charles River Breeding Laboratory (North Wilmington, 

MA), weighing between 230 and 257 grams and approximately 60 days old

Number of animals/sex/dose: 1 male/dose

Route of administration: Oral

Vehicle: Corn oil

Doses/concentrations tested: The test began by dosing at 670 mg/kg, and the dose was raised or 

lowered as needed in accord with the concentration gradients of Deichmann and LeBlanc (1943). 

Dose levels were 12, 17, 26, 40, and 670 mg/kg.

Statistical methods used: Not stated

Test method/conditions remarks: A single oral dose was administered via gavage. A 14-day 

post-dosing observation period was included.

RESULTS

Endpoint value(s): The approximate lethal dose was 26 mg/kg.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

54

Results remarks: Animals dosed with 12 or 17 mg/kg survived. All others died.

CONCLUSIONS

Tetraethyl lead was classified as highly toxic.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. Only one animal was tested per dose level.

REFERENCE

Kennedy, G.L. et al. 1986. Estimation of acute oral toxicity in rats by determination of the 

approximate lethal dose rather than the LD50. J of Appl. Toxicol., 6(3): 145-148.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

55

Study 13: Acute inhalation toxicity study in male rats

Title: Initial submission: Department of Transportation Inhalation Toxicity Studies of 

Tetraethyllead in Rats

The study is: Adequate Not Adequate

The duration of exposure (1 hour) was insufficient to assess inhalation toxicity; the OECDrecommended exposure duration is 4 hours. Additionally, test substance purity was not 

specified.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Described as Haskell #8997 (TEL-Av.)

METHOD

Method/guideline followed: Not specified

Test type: Acute inhalation

GLP compliant?: The study pre-dates GLP.

Year study performed: 1972

Test species: Male ChR-CD rats, initial body weight of 250 grams

Number of animals/sex/dose: 10 males/dose

Route of administration: Inhalation

Vehicle: Air

Doses/concentrations tested: 125, 201, 241, 243, 259, 277, 299, and 435 ppm (0.985, 1.38, 

1.89, 1.91, 2.03, 2.17, 2.35, and 3.4 mg/L)

Statistical methods used: Not specified

Test method/conditions remarks: House air was passed through a fritted bubbler immersed in 

the test material. The resulting vapor mixture was then diluted with air just prior to introduction 

into the chamber. Each exposure duration was 1 hour and utilized 10 rats. Samples for analysis 

were collected by drawing chamber air through a fritted bubbler immersed in iodine 

monochloride solution which has been shown to collect these organic compounds. The resulting 

iodine monochloride-organic lead solutions were analyzed by atomic absorption.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

56

It appears as though a number of different formulations of the test item were tested, although this 

is not entirely clear from the available summary.

RESULTS

Endpoint value(s): The inhalation LC50 value was >125 but <200 ppm.

Results remarks: All rats died at 435, 299, 277, and 241 ppm. Nine rats died at 259 ppm, 5 rats 

died at 201 ppm, and 1 rat died at 243 ppm. There were no mortalities at 125 ppm. Three rats 

that died several days after exposure to 277 ppm were examined for gross pathology, which 

revealed congestion, edema, and hemorrhage in the lungs.

CONCLUSIONS

Tetraethyl lead was classified as highly toxic.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The duration of exposure (1 hour) was insufficient to assess 

inhalation toxicity; the OECD-recommended exposure duration is 4 hours. Additionally, test 

substance purity was not specified.

REFERENCE

TSCATS submission (OTS0571595, Document ID 88-920009938, received 10/27/92). Haskell 

Laboratory, Dupont Chemical Company. 1975. Initial submission: Department of 

Transportation Inhalation Toxicity Studies of Tetraethyllead in Rats with Cover Letter Dated 

101592. 

 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

57

Study 14: Acute inhalation toxicity study in rats

Title: Further studies on the toxicity of some tetra and trialkyl lead compounds

The study is: Adequate Not Adequate

The exposure duration was only 1 hour; the OECD-recommended exposure duration is 4 hours. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Pure sample

METHOD

Method/guideline followed: Not specified

Test type: Acute inhalation

GLP compliant?: The study pre-dates GLP.

Year study performed: 1961

Test species: Adult albino rats (175-230 g)

Number of animals/sex/dose: 12 to 20 rats/group (number per sex not indicated)

Route of administration: Inhalation; 1-hr exposure period

Vehicle: Air

Doses/concentrations tested: 0.095, 0.19, 0.37, 0.77, 1.58, 2.77, 4.84 mg/L (nominal);

0.08, 0.17, 0.34, 0.66, 1.5, 2.64, 3.87 mg/L (analytical mean); approximate dose in mg/kg: 4.8, 

10.2, 20.4, 39.6, 90, 158, and 232 mg/kg, respectively.

Statistical methods used: The LC50 value was calculated according to the method of Weil 

(1952).

Test method/conditions remarks: The 1-hour inhalation exposures were carried out in an allglass chamber. Groups of four rats were exposed head-only to a pure solution of tetraethyl lead 

which was injected into the chamber by a mechanically-driven syringe. Air flow was maintained 

at 50 L/min. 

RESULTS

Endpoint value(s): The 1-hour inhalation LC50 value is 0.85 mg/L, based on mean measured 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

58

concentrations.

Results remarks: 4.84 mg/L: Eight rats died in less than 6 hours and the remaining rats died in 

less than 12 hours. Rats were intensely cyanotic at death.

2.77 mg/L: All rats died in less than 24 hours.

1.58 mg/L: Three rats died in 24 hours and the rest died within 72 hours.

0.77 mg/L: All rats showed excitable sparring behavior; one rat died after 2 days; 2 rats showed 

continuous tremors leading to death in 14 days.

0.37 and 0.19 mg/L: All rats were affected with mild signs of poisoning with slow recovery.

0.095 mg/L: Very mild signs of poisoning, with full recovery.

CONCLUSIONS

Not specified

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The exposure duration was only 1 hour; the OECD-recommended 

exposure duration is 4 hours. 

REFERENCE

Cremer, J.E. and S. Callaway. 1961. Further studies on the toxicity of some tetra and trialkyl 

lead compounds. Brit. J. Ind. Med., 18: 277-282.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

59

Study 15: Acute inhalation toxicity study 

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary lacked sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute inhalation

GLP compliant?: The study pre-dates GLP.

Year study performed: 1973

Test species: Mouse

Number of animals/sex/dose: Not specified

Route of administration: Inhalation

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: The duration of exposure was 7 hours.

RESULTS

Endpoint value(s): The inhalation LCLO value was 0.65 mg/L.

Results remarks: Not specified

CONCLUSIONS

Not specified
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

60

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacked sufficient information to 

assess data adequacy. 

REFERENCE

Primary reference (unavailable): Japanese Journal of Industrial Health, 15: 3. 1973. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

61

Study 16: Acute dermal toxicity study in dogs and guinea pigs

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary lacked sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Acute dermal

GLP compliant?: The study pre-dates GLP.

Year study performed: 1973

Test species: Dogs and guinea pigs

Number of animals/sex/dose: Not specified

Route of administration: Dermal

Vehicle: Not specified

Doses/concentrations tested: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: Not specified

RESULTS

Endpoint value(s): Two dermal LDLO values were provided for dogs: 547 and 830 mg/kg. The 

dermal LDLO for guinea pigs was 995 mg/kg.

Results remarks: Not specified
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

62

CONCLUSIONS

Not specified

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacked sufficient information to 

assess data adequacy. 

REFERENCE

Primary reference (unavailable): Japanese Journal of Industrial Health, 15: 3. 1973. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

63

B-5. REPEATED-DOSE TOXICITY

Summary of endpoint: Inadequate

Study 1: Oral 14-day repeated-dose toxicity study in female mice

Title: Evaluation of tetraethyl lead: Immunology

The study is: Adequate Not Adequate

The study is inadequate to assess repeated-dose toxicity because the exposure duration was only 

14 days; OECD test guidelines specify a minimum exposure period of 28 days. Additionally, no 

LOAEL or NOAEL is identified.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Lot #68573; supplied by the Radian Corporation.

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose/Immunological toxicity

GLP compliant?: Not stated

Year study performed: 1985

Test species: Female B6C3F1 mice from Charles River Breeding Laboratories; 5-6 weeks of 

age, weighing 17 to 19 g.

Number of animals/sex/dose: 8/group

Route of administration: Oral gavage

Vehicle: Corn oil

Exposure duration: 14 days

Doses/concentrations tested: 0, 0.5, 1, or 2 mg/kg

Statistical methods used: Data were tested for homogeneity using the Bartlett’s Chi Square 

Test; for data that were homogeneous, a one-way ANOVA was performed. If the ANOVA 

showed significance, then a Dunnett’s Multiple Range T-test was performed. If the data were 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

64

non-homogeneous, a non-parametric ANOVA, Kruskal-Wallis test, and Wilcoxon Rank Sum 

Test were performed.

Test method/conditions remarks: Mice received oral gavage doses daily for 14 consecutive 

days. Body weights were measured on days 1, 8, and 15 of the study. Animals were observed at 

the time of dosing for any clinical signs. Mice were necropsied 24 hours after the last exposure. 

Organ weights (brain, liver, lung, spleen, thymus, and kidneys) were measured. Gross and 

histopathological analyses were performed on selected organs (liver, spleen, lungs, thymus, 

mesenteric lymph nodes, and kidneys). Hematological and clinical chemistry analyses were 

performed.

Immunological tests included spleen IgM antibody response to a T-dependent antigen (sheep red 

blood cells), mixed lymphocyte response to DBA/2 mouse spleen cells, delayed hypersensitivity 

response to keyhole limpet cyanin, and serum complement levels.

RESULTS

Endpoint value(s): None specified

Results remarks: The high dose produced nervous system toxicity (tremors, beginning on day 5 

of exposure and progressing to paralysis). Dose-dependent decreases in erythrocyte count, 

hematocrit, hemoglobin, and peripheral blood leukocytes (lymphocytes and neutrophils) were 

seen. There was a slight but significant increase in BUN. Histopathological evaluation was 

negative with respect to lymphoid tissue, but a mild chronic hepatitis was seen at 1 and 2 mg/kg. 

Body weights decreased in a dose-dependent fashion, reaching statistical significance at 2 mg/kg. 

No significant effects were observed on organ weights.

There were no effects on IgM antibody response, mixed lymphocyte response, and 

hypersensitivity response. Serum complement levels, as measured by total hemolytic activity, 

were decreased dose-dependently.

CONCLUSIONS

None specified

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The exposure duration was only 14 days; OECD test guidelines 

specify a minimum exposure period of 28 days. Additionally, no LOAEL or NOAEL is 

identified.

REFERENCE

Luster. M.I. 1986. Evaluation of tetraethyl lead: Immunology. Report to National Toxicology 

Program. June 1, 1986. NIEHS, RTP. Contract No. ES 55094.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

65

Study 2: Oral 13-week repeated-dose toxicity study in male rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary lacks sufficient information to assess data adequacy. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: Not stated

Year study performed: 1987

Test species: Male Sprague-Dawley rats

Number of animals/sex/dose: Not indicated 

Route of administration: Oral gavage

Vehicle: Not indicated

Exposure duration: 5 times/week for 13 weeks

Doses/concentrations tested: 0, 0.2, or 2 mg/kg

Statistical methods used: Not stated

Test method/conditions remarks: Not specified

RESULTS

Endpoint value(s): None specified

Results remarks: Symptoms included decreased food intake and body weight, hyperexcitability, 

irritability, tremor, and coma. Pathological changes included brain and spinal cord lesions and 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

66

eosinophile inclusions in the proximal renal tubule.

CONCLUSIONS

None specified.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary lacks sufficient information to 

assess data adequacy. 

REFERENCE

Primary reference (unavailable): Franklin C.A. et al., 1987. Trace Subst. Environ. Health 21: 

286-296.

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

67

Study 3: Oral 21-week repeated-dose toxicity study in rats

Title: Tetraethyllead and tetramethyllead

The study is: Adequate Not Adequate

No endpoint value was specified in the report, and parameters investigated were non-standard. 

Also, the study report did not specify which effects were seen at which concentrations or the 

statistical significance of effects. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Dissolved in peanut oil to yield a concentration of 1.813 mg/mL; toluene added (as a 

stabilizer) at 0.321 mL per 100 mL peanut oil

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1964

Test species: Charles River cesarean-derived albino rats (22 days old)

Number of animals/sex/dose: 6/sex/dose for treated groups; 8/sex/dose for control group

Route of administration: Oral gavage

Vehicle: Peanut oil/toluene

Exposure duration: 21 weeks

Doses/concentrations tested: 0, 0.0017, or 0.17 mg/kg

Statistical methods used: Not specified

Test method/conditions remarks: Animals were housed in stainless steel wire cages and 

observed for 9 days pre-dosing. Pre-dosing hematological data were obtained. Four male and 

for female rats were sacrificed for histologic examination pre-dosing. 

Animals received daily oral gavage doses of the test item 5 times/week to a total of 100 doses. 

All rats were sacrificed during the first or second day after administration of the last dose. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

68

Terminal measurements included body weight, organ weights, lead content in tissues, and 

histopathology.

RESULTS

Endpoint value(s): None specified

Results remarks: Peripheral hyperemia was noted in high-dose rats. Irritability of the animals 

increased as the experiment proceeded. No adverse effects on terminal body weight were 

observed. Some slight organ weight changes were seen with respect to controls, but no statistical 

analysis was provided. 

Pathological changes included swollen friable liver with hypotrophy and hypoplasia of the 

hepatocytes; epithelial hyperplasia of the thyroid gland; fatty degeneration of the thymus; 

hypertrophy of the parathyroids; hyperemia and edema of the central nervous system; neural 

degeneration; and pulmonary edema.

The liver and kidneys had the highest accumulation of lead, relative to other organs.

CONCLUSIONS

Tetraethyl lead caused significant lesions in various organs and selective distribution of lead in 

tissues.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. No endpoint value was specified in the report, and parameters 

investigated were non-standard. Also, the study report did not specify which effects were seen at 

which concentrations or the statistical significance of effects. 

REFERENCE

Schepers, G.W.H. 1964. Tetraethyllead and tetramethyllead. Arch Env Health, 8: 277-295. 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

69

Study 4: Oral repeated-dose toxicity study in rats

Title: None specified

The study is: Adequate Not Adequate

The IUCLID summary, which was mostly in a language other than English, lacked sufficient 

information to assess data adequacy.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: Not specified

Year study performed: 1980

Test species: Rats

Number of animals/sex/dose: Not specified

Route of administration: Oral gavage

Vehicle:

Exposure duration: Not specified

Doses/concentrations tested: 5 and 10 mg/kg

Statistical methods used: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): None specified

Results remarks: The IUCLID summary of the study was mostly in a language other than 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

70

English.

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which was mostly in a language 

other than English, lacked sufficient information to assess data adequacy.

The IUCLID summary specified a reliability rating of 2.

REFERENCE

Primary reference (unavailable): Seawright A.A. et al. 1980. In Holmstedt et al. Mechanism of 

Toxicity and Hazard Evaluation. Elsevier/North Holland, 71-74. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

71

Study 5: Oral 6-month repeated-dose study in rhesus monkeys

Title: Chronic oral administration of alkyl lead solutions to the rhesus monkey

The study is: Adequate Not Adequate

Only one test concentration was employed, as well as a non-standard test species and insufficient 

number of animals per treatment group.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Not specified

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1979

Test species: Rhesus monkeys (Macaca mulatta), weighing 2.1-3.3 kg; wild-caught

Number of animals/sex/dose: 2/sex/dose

Route of administration: oral gavage

Vehicle: Toluene/ethanol

Exposure duration: 6 months

Doses/concentrations tested: 0 (toluene/ethanol vehicle control), 6 μg Pb/kg/day

Statistical methods used: Analysis of variance and Newman-Keuls’ multiple comparison 

procedure were used.

Test method/conditions remarks: Each monkey was housed in a wall-mounted cage, in a room 

maintained at a temperature of 22 oC in normal daylight conditions. Monkeys were given daily 

gavage doses of 4 mL/kg of the test substance for 6 months at a dose level equivalent to 6 μg 

Pb/kg/day. Clinical signs and food consumption were monitored daily throughout. Body 

weights were measured weekly. Neurological and ophthalmoscopic examinations were made 

throughout. Hematological and blood biochemical examinations were made before and after 13 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

72

and 26 weeks of treatment. Twenty-four hour urine samples were examined. Lead content of 

blood, urine, and feces was estimated at weekly intervals for the first 8 weeks, and thereafter at 

2-week intervals. Post-mortem examinations included weighing of brain, pituitary, heart, lungs, 

liver, spleen, pancreas, thymus, prostate or uterus, kidneys, adrenals, and gonads, and a wide 

range of tissues were prepared for histological examination. Samples of all major tissues were 

analyzed for lead content.

RESULTS

Endpoint value(s): None specified

Results remarks: There were no effects of treatment except for elevated blood, urine, and feces 

lead levels, and elevated lead in tissues.

CONCLUSIONS

The administration of tetraethyl lead to rhesus monkeys for long periods was without effect on 

the clinical, behavioral, and pathological parameters monitored.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. Only one test concentration was employed, as well as a nonstandard test species and insufficient number of animals per treatment group.

REFERENCE

Heywood R. et al. 1979. Chronic oral administration of alkyl lead solutions to the rhesus 

monkey. Toxicol. Lett., 4: 119-125.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

73

Study 6: Inhalation repeated-dose toxicity study in rats

Title: Inhalation of tetramethyllead and tetraethyl lead

The study is: Adequate Not Adequate

The exposure regimen and parameters investigated were non-standard. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 99.3% purity; supplied by the Ethyl Corporation.

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1963

Test species: CFN rats obtained from Carworth Farms, NY; 3 months old at test initiation

Number of animals/sex/dose: 10/sex/concentration for control group; 5/sex/concentration for 

treated groups

Route of administration: Inhalation

Vehicle: Air

Exposure duration: 0 and 12 mg/m3

: 150 7-hour exposure periods; 22 mg/m3:14 7-hour 

exposure periods; 46 mg/m3: 5 7-hour exposure periods.

Doses/concentrations tested: 0, 12, 22, or 46 mg/m3

Statistical methods used: Not stated

Test method/conditions remarks: The vapor was generated by bubbling dry nitrogen through 

the liquid contained in a glass cylinder fitted with sintered disks for dispersing the gas through 

the liquid. Rats were confined in 32-liter glass bell jars. The vapor and supplemental air were 

introduced at a rate of 32 liters/minute. Three or more samples of test atmosphere were taken 

during each 7-hour period of exposure.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

74

Tissues and excreta were analyzed for lead content, and pathological changes in viscera were 

investigated.

RESULTS

Endpoint value(s): None specified

Results remarks: At 22 mg/m3, 4/5 males and 5/5 females died. At 46 mg/m3, 3/5 males and 

5/5 females died. There were no mortalities at 0 and 12 mg/m3. Clinical signs included 

irritability when handled, uncoordinated movements, combativeness, convulsions, and coma 

followed by death. Tetraethyl lead was found to be concentrated mostly in the lungs, liver, and 

kidneys; the lead content in the brain was 10 μg/g (exposure concentration not specified). 

Pathological findings included brain damage (neocortex, medulla oblongata, and brain stem), 

including cell shrinkage, vacuolization, and extended neuronal necroses with glioses in both 

hemispheres of the brain; swollen hepatocytes; swelling and necrosis of the proximal renal 

tubules; and hemorrhages and inflammation of the lungs.

CONCLUSIONS

Tetraethyl lead was more toxic to rats than tetramethyl lead.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The exposure regimen and parameters investigated were nonstandard. 

REFERENCE

Davis, R.K. et al. 1963. Inhalation of tetramethyllead and tetraethyllead: A comparison of the 

effects in rats and dogs. Arch. Environ. Health, 6: 473-479.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

75

Study 7: Inhalation repeated-dose toxicity study in dogs

Title: Inhalation of tetramethyllead and tetraethyllead

The study is: Adequate Not Adequate

The exposure regimen, test species, and parameters investigated were non-standard; additionally, 

only one or two animals were tested per concentration. 

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 99.3% purity; supplied by the Ethyl Corporation.

METHOD

Method/guideline followed: Not specified

Test type: Repeated-dose toxicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1963

Test species: Purebred beagle dogs, obtained from Cherry Hill Kennels, OH or Cornell Dog 

Farm, NY; initial weight: 5.9 to 13.2 kg.

Number of animals/sex/dose: 0 and 12 mg/m3

: 2/concentration, 22 and 42 mg/m3: 

1/concentration; sex not specified 

Route of administration: Inhalation

Vehicle: Air

Exposure duration: 0 mg/m3

: more than 130 7-hour exposures; 12 mg/m3: 24 and 29 7-hour 

exposure periods for a smaller (5.9 kg) and larger (10.2 kg) dog, respectively; 22 mg/m3:30 7-

hour exposure periods; 42 mg/m3: 7 7-hour exposure periods.

Doses/concentrations tested: 0, 12, 22, or 42 mg/m3

Statistical methods used: Not stated

Test method/conditions remarks: The vapor was generated by bubbling dry nitrogen through 

the liquid contained in a glass cylinder fitted with sintered disks for dispersing the gas through 

the liquid. Dogs were subjected to the inhalation of the test item in insulated 600-liter plywood 

chambers which were supplied with air at a rate of 600 liters/minute; the air was dehumidified to 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

76

50% relative humidity and the temperature was 77 to 81 oF. Three or more samples of test 

atmosphere were taken during each 7-hour exposure period.

Tissues and excreta were analyzed for lead content, and pathological changes in viscera were 

investigated.

RESULTS

Endpoint value(s): None specified

Results remarks: It appears that all dogs died, although this is not explicitly stated in the report. 

Clinical signs included tremors, muscle twitching, hyperactivity, and coma followed by death. 

Tetraethyl lead was found to be concentrated mostly in the liver and kidneys. The lead 

concentration in the brain was 2.0 μg/g (exposure concentration not specified). Pathological 

findings included brain damage (neocortex, medulla oblongata, and brain stem), including cell 

shrinkage, vacuolization, and extended neuronal necroses with glioses in both hemispheres of the 

brain; swollen hepatocytes; swelling and necrosis of the proximal renal tubules; and hemorrhages 

and inflammation of the lungs. 

CONCLUSIONS

Tetraethyl lead was more toxic to dogs than to rats.

STUDY RELIABILITY

Klimisch code = [3]. Invalid. The exposure regimen, test species, and parameters investigated 

were non-standard; additionally, only one or two animals were tested per concentration. 

REFERENCE

Davis, R.K. et al. 1963. Inhalation of tetramethyllead and tetraethyllead: A comparison of the 

effects in rats and dogs. Arch. Environ. Health, 6: 473-479.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

77

B-6. GENETIC TOXICITY – GENE MUTATION

Summary of endpoint: Adequate for gene mutations; inadequate for chromosomal aberrations

Study 1: Mutagenicity study in Salmonella

Title: Salmonella mutagenicity tests: II. Results from the testing of 270 chemicals

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: Supplier: ICN Pharmaceuticals

METHOD

Method/guideline followed: According to Haworth et al. (1983)

Test type: Preincubation assay

GLP compliant?: Not specified

Year study performed: 1986

Test species: Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537, obtained 

from Dr. Bruce Ames

Metabolic activation: With and without

Concentrations tested: 0, 1.0, 3.3, 10.0, 33.3, 100 μg/plate with 95% ethanol as a solvent; 0, 10, 

33, 100, 333, 1000 μg/plate with DMSO as a solvent

Statistical methods used: The chemical was considered to be mutagenic if there was a doserelated, reproducible increase in the number of revertants over the background level, even if the 

increase was less than two-fold. If no increase in the number of revertants was observed, the 

chemical was considered to be non-mutagenic. The response was considered to be equivocal if 

there was an absence of a clear-cut dose-related increase in revertants, if the dose-related 

increases in revertants were not reproducible, or if the response was of insufficient magnitude to 

support a determination of mutagenicity.

Test method/conditions remarks:

Preparation of S-9 fraction: Male Sprague-Dawley rats and male Syrian hamsters were routinely 

used for the preparation of the liver fractions. Aroclor 1254 (200 mg/mL in corn oil) was

administered i.p. at 500 mg/kg 5 days prior to decapitation or cervical dislocation. The animals 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

78

were deprived of food for 12-24 hours immediately preceding death; otherwise food and water 

were provided ad libitum. The livers were removed aseptically, washed in ice-cold 0.15 M KCl, 

and minced and homogenized (3 mL of 0.15 M KCl per g of wet tissue). The S-9 fraction was 

obtained by centrifugation of the liver homogenate for 10 min at 9,000 g at 4°C. The S-9 fraction 

was dispensed into freezing ampoules and stored in a -70°C freezer, or in liquid nitrogen. The S9 mix was prepared immediately prior to each assay and consisted of the following, per 

milliliter: S-9 fraction, 0.10 mL; 0.04 M MgC12, 0.02 mL; 1.65 M KC1, 0.02 mL; 0.04 M Pnicotinamide adenine dinucleotide phosphate (NADP), 0.10 mL; 0.05 M glucose-6-phosphate, 

0.10 mL; 1.0 M NaH2PO4, (pH 7.4), 0.10 mL; and distilled water, 0.56 mL. 

Preincubation assay: All chemicals were assayed for mutagenicity in the preincubation assay. 

To each of 13 x 100-mm test tubes maintained at 37 oC were added in the following order: 0.5 

mL of S-9 mix or 0.1 M PO4 buffer (pH 7.4), 0.05 mL of the overnight culture, and 0.05 mL of 

solvent or chemical dilution. The mixture was mixed and allowed to incubate without shaking at 

37 oC for 20 min, at which time 2.5 or 2.0 mL of molten (45°C) top agar supplemented with 0.5 

rnM L-histidine and 0.5 mM D-biotin were added. The contents of the tubes were mixed and 

poured onto 25 mL of minimal glucose bottom agar in 15 x 100-mm plastic petri dishes. When 

the top agar had solidified, the plates were inverted and incubated at 37 oC for 48 hours. 

Concurrent solvent and positive controls were tested with and without the metabolic activation 

systems. At least five dose levels were tested, with three plates per dose level. All assays were 

repeated (as described above) no less than 1 wk after completion of the initial test.

Controls: The following mutagens were used as concurrent positive controls in the absence of 

activation: sodium azide for TA1535 and TA100, 4-nitro-o-phenylenediamine for TA98, and 9-

aminoacridine for TA97 and TA1537; 2-aminoanthracene was used with all strains with hamster 

and rat liver metabolic activation systems. 

RESULTS

Genotoxicity result: Negative with and without metabolic activation

Cytotoxic concentration (if applicable): Slight cytotoxicity at 100 μg/plate in 95% ethanol

Results remarks: Positive and negative controls responded appropriately

CONCLUSIONS

None specified

STUDY RELIABILITY

Klimisch code = [1]. Reliable without restrictions.

REFERENCE
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

79

Mortelmans et al. 1986. Salmonella Mutagenicity Tests: II. Results From the Testing of 270 

Chemicals. Environmental Mutagenesis, 8 (Suppl. 7): 1-119.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

80

B-7. REPRODUCTIVE TOXICITY 

Study 1: Reproductive toxicity study in mice

Summary of endpoint: Adequate. Although the reproductive study of tetraethyl lead in mice 

(Study 1) is not adequate, sufficient, reliable studies on the neurodevelopmental toxicity of lead, 

as reviewed in the U.S. EPA document Air Quality Criteria for Lead, Vol I of II (U.S. EPA 

2006), exist to support a determination of adequacy for the reproductive toxicity endpoint based 

on a weight-of-evidence approach.

Reference: U.S. EPA 2006. Air Quality Criteria for Lead. Volume I of II. EPA/600/R5/144aF, October 2006. Available on-line at 

http://cfpub.epa.gov/ncea/cfm/recordisplay.cfm?deid=158823#Download

Title: Not specified

The study is: Adequate Not Adequate

The IUCLID summary, which is mostly in a language other than English, does not contain 

sufficient information to assess data adequacy.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: None specified

METHOD

Method/guideline followed: Not specified

Test type: Fertility study

GLP compliant?: The study pre-dates GLP.

Year study performed: 1971

Test species: Mouse

Number of animals/sex/dose: Not specified

Route of administration: Oral feed

Exposure duration: Not specified

Doses/concentrations tested: 6.48, 32 mg/kg
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

81

Vehicle: Not specified

Statistical methods used: Not specified

Test method/conditions remarks: The IUCLID summary of the study was mostly in a language 

other than English.

RESULTS

Endpoint value(s): Not specified

Results remarks: The IUCLID summary of the study was mostly in a language other than 

English.

CONCLUSIONS

The IUCLID summary of the study was mostly in a language other than English.

STUDY RELIABILITY

Klimisch code = [4]. Not assignable. The IUCLID summary, which is mostly in a language 

other than English, does not contain sufficient information to assess data adequacy.

REFERENCE

Primary reference (unavailable): Kennedy, G.L. and D. Arnold. 1971. Newsletter Environ Mut 

Soc 5: 370. 

Secondary reference: IUCLID Dataset for CAS No. 78-00-2, available at 

http://ecb.jrc.ec.europa.eu/esis/
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

82

B-8. DEVELOPMENTAL TOXICITY 

Summary of endpoint: Adequate

Study 1: Developmental toxicity study in mice

Title: Teratogenic evaluation of lead compounds in mice and rats

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Houston Chemical Company, Freeport, TX

METHOD

Method/guideline followed: Not specified

Test type: Teratogenicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1975

Test species: CD-1 random-bred albino female mice, impregnated at the Charles River Breeding

Laboratories, Wilmington, MA

Number of animals/sex/dose: 20

Route of administration: Oral gavage

Exposure duration: Gestation day 5 to 15

Doses/concentrations tested: 0, 0.01, 0.1, 1.0, and 10 mg/kg

Vehicle: Corn oil

Statistical methods used: Not specified

Test method/conditions remarks: Animals received oral gavage doses daily from gestation day 

5 through 15. Animals were weighed daily. Due to the toxicity observed at 10 mg/kg, treatment 

at this level was discontinued after 3 daily doses. Animals were sacrificed by cervical 

dislocation on day 18 of gestation and the uterine horns were examined for implantation sites, 

resorption sites, and viable fetuses. A complete examination of each fetus was conducted and, 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

83

following removal of excess amniotic fluid, each was weighed. Fetuses were divided into two 

equal groups (randomly assigned in each litter) for evaluation of skeletal development or of 

internal development.

RESULTS

Endpoint value(s): None specified, but the LOAEL appears to be 1 mg/kg for systemic toxicity 

(based on weight loss) and for developmental toxicity (increased number of resorptions and 

decreased fetal growth). The NOAEL for teratogenicity appears to be 10 mg/kg, the highest 

dose level tested.

Results remarks: Animals given up to 0.10 mg/kg showed no clinical signs and displayed 

normal weight gains during pregnancy. Mice treated with 1 mg/kg gained 25% less weight 

during gestation than controls. Mice exhibited hypoactivity, tremors, and convulsions along with 

severe body weight loss after 3 daily doses of 10 mg/kg, and subsequent dosing was 

discontinued.

The number of pregnancies was sharply reduced in mice given 10 mg/kg (5 pregnant females, 

compared to 16 in control group). Many of these high-dose females were observed to be full of 

brown liquid without any traces of fetal tissues. The mean number of resorption sites per female 

was 0.8, 0.7, 0.9, 2.1, and 1.6 at 0 (vehicle control), 0.01, 0.1, 1.0, and 10 mg/kg, respectively. 

The mean number of fetuses/litter was 10.9, 11.7, 11.9, 10.2, and 10.0 at 0 (vehicle control), 

0.01, 0.1, 1.0, and 10 mg/kg, respectively. 

No evidence for a teratogenic response to lead was obtained from the gross examination of mice. 

Three of 362 fetuses (0.8%) displayed external anomalies, consisting of one exencephaly, one 

runt, and one cleft palate. Three abnormal fetuses, all displaying torsion of the rear limbs, were 

recovered from one litter at 0.10 mg/kg. Skeletal examination revealed slight delays in 

ossification in fetuses from females exposed to 1 or 10 mg/kg. Internal evaluation revealed no

significant effects.

Growth of fetuses, as indicated by terminal body weights, was reduced at doses ≥ 1.0 mg/kg.

CONCLUSIONS

It is concluded that no teratogenic response is produced in mice following in utero exposure to 

maximally tolerated doses of tetraethyl lead.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. Apparently, no statistical evaluation of 

teratogenicity data was performed.

REFERENCE

Kennedy, G.L. et al. 1975. Teratogenic evaluation of lead compounds in mice and rats. Food 
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

84

Cosmet. Toxicol., 13: 629-632.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

85

Study 2: Developmental toxicity study in rats

Title: Teratogenic evaluation of lead compounds in mice and rats

The study is: Adequate Not Adequate

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: 98% pure, supplied by Houston Chemical Company, Freeport, TX

METHOD

Method/guideline followed: Not specified

Test type: Teratogenicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1975

Test species: COBS random-bred albino female rats, impregnated at the Charles River Breeding

Laboratories, Wilmington, MA

Number of animals/sex/dose: 20

Route of administration: Oral gavage

Exposure duration: Gestation day 6 to 16

Doses/concentrations tested: 0, 0.01, 0.1, 1.0, and 10 mg/kg

Vehicle: Corn oil

Statistical methods used: Not specified

Test method/conditions remarks: Animals received oral gavage doses daily from gestation day 

6 through 16. Animals were weighed daily. Due to the toxicity observed at 10 mg/kg, treatment 

at this level was discontinued after 3 daily doses. Animals were sacrificed by cervical 

dislocation on day 20 of gestation and the uterine horns were examined for implantation sites, 

resorption sites, and viable fetuses. A complete examination of each fetus was conducted and, 

following removal of excess amniotic fluid, each was weighed. Fetuses were divided into two 

equal groups (randomly assigned in each litter) for evaluation of skeletal development or of 

internal development.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

86

RESULTS

Endpoint value(s): None specified, but the LOAEL appears to be 1 mg/kg for systemic toxicity 

(based on weight loss) and for developmental toxicity (increased number of resorptions and 

decreased fetal growth). The NOAEL for teratogenicity appears to be 10 mg/kg, the highest 

dose level tested.

Results remarks: Animals given up to 0.10 mg/kg showed no clinical signs and displayed 

normal weight gains during pregnancy. Rats treated with 1 mg/kg gained 70% less weight 

during gestation than controls. Rats exhibited hypoactivity, tremors, and convulsions along with 

severe body weight loss after 3 daily doses of 10 mg/kg, and subsequent dosing at this dose level 

was discontinued.

The number of pregnancies was sharply reduced in rats given 10 mg/kg (7 pregnant females, 

compared to 16 in control group). Many of these high-dose females were observed to be full of 

brown liquid without any traces of fetal tissues. The mean number of resorption sites per female 

was 0.8, 0.7, 0.9, 2.3, and 1.2 at 0 (vehicle control), 0.01, 0.1, 1.0, and 10 mg/kg, respectively. 

The mean number of fetuses/litter was 11.5, 11.9, 11.9, 10.5, and 9.9 at 0 (vehicle control), 0.01, 

0.1, 1.0, and 10 mg/kg, respectively. 

One fetus at 0.01 mg/kg was non-viable, and one fetus at 0.10 mg/kg had malrotation of a 

hindlimb. All other fetuses appeared normal, indicating a lack of a teratogenic response. No 

internal findings were observed in fetuses. Upon skeletal examination, delayed ossification was 

seen in offspring at 10 mg/kg.

Growth of fetuses, as indicated by terminal body weights, was reduced at doses ≥ 1.0 mg/kg.

CONCLUSIONS

It is concluded that no teratogenic response is produced in rats following in utero exposure to 

maximally tolerated doses of tetraethyl lead.

STUDY RELIABILITY

Klimisch code = [2]. Reliable with restrictions. Apparently, no statistical evaluation of 

teratogenicity data was performed.

REFERENCE

Kennedy, G.L. et al. 1975. Teratogenic evaluation of lead compounds in mice and rats. Food 

Cosmet. Toxicol., 13: 629-632.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

87

Study 3: Developmental toxicity study in rats

Title: Effects of organolead compounds on rat embryonic and fetal development

The study is: Adequate Not Adequate

The duration of exposure was insufficient; the OECD-recommended duration extends from 

gestation day 6 to 15 for rodents.

TEST SUBSTANCE 

Identity: Tetraethyl lead, CAS No. 78-00-2

Remarks: >99% pure; provided by The Ethyl Corporation, Ferndale, MI

METHOD

Method/guideline followed: Not specified

Test type: Developmental toxicity

GLP compliant?: The study pre-dates GLP.

Year study performed: 1972

Test species: Pregnant Simonsen Sprague-Dawley rats

Number of animals/sex/dose: 3 to 8/group

Route of administration: Oral gavage

Exposure duration: Gestation day 9-11 or 12-15

Doses/concentrations tested: 0, 7.5, 15, or 30 mg/kg

Vehicle: Corn oil

Statistical methods used: Statistical analyses were made by binomial expansion. Quantitative 

data were analyzed by ANOVA and individual means compared using the group t-test.

Test method/conditions remarks: Animals received oral gavage doses daily from gestation 

days 9 through 11, or gestation days 12 through 14. Rats were sacrificed on gestation day 22 and 

the total number and position of live, dead, or resorbed fetuses were recorded. Fetuses were 

weighed and examined for gross malformations; the transumbilical and crown-rump lengths 

were measured. One half of each litter was placed in alcohol and the other half in Bouin 

solution. Soft tissue and skeletal examinations for anomalies were performed.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

88

RESULTS

Endpoint value(s): None specified, but the LOAEL appears to be 7.5 mg/kg for systemic 

toxicity (based on weight loss) and for developmental toxicity (based on decreased fetal body 

weights). 

Results remarks: Clinical signs of toxicity ranged from hyperexcitability and weight loss at 

lower levels to tremors, aggressiveness, spasticity, and paralysis at higher dose levels. The 

highest dose level resulted in death of all dams. At 30 mg/kg, 5/5 rats dosed on days 9 through 

11 died, and 4/4 rats dosed on days 12-14 died. One rat dosed at 15 mg/kg on days 12-14 also 

died.

An increased number of resorbed fetuses was observed at the highest dose level allowing 

survival (15 mg/kg) with tetraethyl lead administered on days 9, 10, and 11 of gestation. 

Retardation of fetal growth was also evident. Body weights of fetuses were significantly 

decreased, compared with controls, at 7.5 and 15 mg/kg (for both dosing periods). Changes in 

transumbilical and crown-rump length and the length of the long bones in the fetus paralleled 

changes in body weight. Growth-retarded fetuses were otherwise normal in external appearance. 

No gross malformation was observed in any control or treated fetus. There were no gross 

deformities of skeletal structures and no significant soft tissue malformations.

CONCLUSIONS

It was concluded that the observed embryotoxic and fetal toxic effects were secondary to the 

maternal toxicity rather than a direct effect of tetraethyl lead on the developing rat.

STUDY RELIABILITY

Klimisch code = [3]. Not reliable. The duration of exposure was insufficient; the OECDrecommended duration extends from gestation day 6 to 15 for rodents. Additionally, data for 

transumbilical and crown-rump length were not provided.

REFERENCE

McClain, R.M. and B.A. Becker. 1972. Effects of organolead compounds on rat embryonic and 

fetal development. Toxicol. Appl. Pharmacol., 21: 265-274.
Draft HPV4 Chemical Data Adequacy Review CAS No. 78-00-2

89

B-9. CAS No. 78-00-2: Summary of Endpoints

Endpoint Study Information 

Available

Acceptable Data Gap

Aquatic Effects

Acute Toxicity to Fish Yes Yes No

Acute Toxicity to Daphnia Yes1 Yes No

Toxicity to Aquatic Plants Yes No Yes

Health Effects

Acute Toxicity Yes Yes No

Repeated-Dose Toxicity Yes No Yes

Gene Mutation Yes Yes No

Chromosomal Aberrations No - Yes

Reproductive Toxicity Yes2 Yes No

Developmental Toxicity Yes Yes No

1

No studies of plumbane, tetraethyl on Daphnia were located; however, three studies were 

located on other aquatic invertebrate species. These studies tended to indicate high toxicity to 

aquatic invertebrates.

2 Although the reproductive study of tetraethyl lead in mice (Study 1) is not adequate, sufficient, 

reliable studies on the neurodevelopmental toxicity of lead, as reviewed in the U.S. EPA 

document Air Quality Criteria for Lead, Vol I of II (U.S. EPA 2006), exist to support a 

determination of adequacy for the reproductive toxicity endpoint based on a weight-of-evidence 

approach.

Reference: U.S. EPA 2006. Air Quality Criteria for Lead. Volume I of II. EPA/600/R5/144aF, October 2006. Available on-line at 

http://cfpub.epa.gov/ncea/cfm/recordisplay.cfm?deid=158823#Download