Phenyl phosphorodichloridate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 1997-05-30 to 1997-07-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

1996-02-22

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phenyl phosphorodichloridate

EC Number:

212-220-6

EC Name:

Phenyl phosphorodichloridate

Cas Number:

770-12-7

Molecular formula:

C6H5Cl2O2P

IUPAC Name:

phenyl dichlorophosphate

Test material form:

liquid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

10% liver S9 in standard co-factor

Test concentrations with justification for top dose:

preliminary test: 0, 50, 150, 1500 and 5000 µg/plate
Main test: 0, 50, 150, 500, 1500 and 5000 µg/plate.

5000 µg/plate is the maximum recommended dose (No visible reduction in the growth of the bacterial lawn.

Vehicle / solvent:

water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

mitomycin C

other: 2-Aminoanthracene and 1,8-dihydroxyanthroquinone (danthron) in the experiment with S9-mix

Details on test system and experimental conditions:

See any other information on materials and methods incl.tables

Evaluation criteria:

See any other information on materials and methods incl.tables

Statistics:

Dunnett's method of linear regression

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Preliminary toxicity study

The mean number of revertant colonies for the toxicity assay report in the table below

Strain	Dose (µg/plate)
	0	50	150	500	1500	5000
TA100	114	123	107	113	108	98

Experiment 1 without metabolic activation

 

With or witjout metabolic activation	Concentration (µg/plate)	Number of revertant (number of colonies per plate)
		Base-pair substitution type						Frameshift type
		TA100		TA1535		TA102		TA98		TA1537
-	0	83		14		215		20		9
			(78)		(17)		(224)				(9)
		74		18		220		10		12
			4.5		3.1		12.1				3.0
		78		20		238		13		6
-	50	79		13		220		22		10
			(78)		(15)		(224)		(14)		(11)
		72		19		211		23		7
			5.1		3.5		14.8		5.1		4.6
		82		13		240		19		16
-	150	79		15		230		12		8
			(77)		(16)		(223)		(21)		-10)
		71		18		226		14		12
			4.9		1.5		8.9		2.1		2.1
		80		16		213		20		11
-	500	71		20		249		16		10
			(75)		(17)		(230)		(15)		(10)
		82		18		232		23		14
			5.9		3.1		19.6		4.2		4
		73		14		210		17		6
-	1500	81		11		234		13		7
			(76)		(13)		(226)		(19)		(10)
		75		17		220		11		13
			5.0		3.8		7.4		3.8		3.1
		71		10		223		21		11
-	5000	74		13		236		10		16
			(77)		(14)		(226)		(15)		(12)
		86		17		229		15		11
			7.6		3.1		11.8		4.5		3.2
		72		11		213		19		10
Positive control	Name	ENNG		ENNG		MMC		4NQO		9AA
S9-Mix	Concentration (µg/plate)	3		5		0.5		0.2		80
-	Number of colonies per plate	890		791		529		247		874
			(863)		(825)		(576)		(218)		(800)
		797		882		618		199		802
			57.1		49.9		44.8		25.5		75.0
		901		801		582		208		724

ENNG: N -ethy 1- N' -nitro- N -n itrosoguanidine

MMC: Mitomycin C

4NQO:4-nitroquinoline-1-oxide

9AA: 9-aminoacridine

Experiment 1 with metabolic activation

 

With or witjout metabolic activation	Concentration (µg/plate)	Number of revertant (number of colonies per plate)
		Base-pair substitution type						Frameshift type
		TA100		TA1535		TA102		TA98		TA1537
+	0	86		11		216		10		7
			(80)		(14)		(227)		(15)		(11)
		73		13		235		21		11
			6.7		3.1		9.7		5.6		3.5
		82		17		229		14		14
+	50	90		21		242		14		10
			(84)		(17)		(233)		(18)		(12)
		89		19		220		23		10
			9.5		5.3		1.4		5.0		2.9
		73		11		236		13		15
+	150	91		15		221		15		12
			(78)		(16)		(217)		(18)		(10)
		71		13		221		23		9
			11.0		3.6		6.4		4.6		2.1
		73		20		210		15		8
+	500	79		10		231		17		13
			(80)		(12)		(226)		(16)		(8)
		85		13		217		12		2
			4.2		2.1		7.6		3.2		2.1
		77		14		229		18		9
+	1500	80		10		226		15		6
			(77)		(13)		(219)		(16)		(6)
		73		11		210		21		7
			3.6		4.4		8.3		4.6		0.6
		78		18		222		12		6
+	5000	76		19		222		12		7
			(78)		(18)		(221)		(15)		(8)
		80		20		215		18		11
			2.1		2.6		5.6		3.1		2.6
		79		15		226		14		6
Positive control	Name	2AA		2AA		DANTHRON		2AA		2AA
S9-Mix	Concentration (µg/plate)	1		2		10		0.5		2
+	Number of colonies per plate	1045		325		510		386		321
			(1046)		(302)		(543)		(312)		(329)
		1121		288		503		256		350
			74.0		19.9		62.7		65.9		18.7
		973		294		615		292		315

2AA: 2-aminoanthracene

DANTHRON: 1,8-dihydroxyanthraquinone

Applicant's summary and conclusion

Conclusions:

The phenyl dichlorophosphate was considered to be non-mutagenic under the conditions of this test

Executive summary:

 Salmonella typhimurium strains T A 1535, T A 1537, T A 102, T A98 and T A 100 were treated with the test material using the Ames plate incorporation method at five dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard co-factors). The dose range was determined in a preliminary toxicity assay and was 50 to 5000 Jig/plate in the first

experiment. The experiment was repeated on a separate day using the same dose range as experiment 1, fresh cultures of the bacterial strains and fresh test material formulations. The method used conforms with the OECD Guidelines for the Testing of Chemicals, Protocol No. 471 and also with Method B14 in Commission Directive 92/69/EEC.

 

The vehicle (sterile distilled water) control plates produced counts of revertant colonies within the normal range.

 

All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without the metabolising system.

 

The test material caused no visible reduction in the growth of the bacterial lawn at any of the dose levels to any of the strains of Salmonella tested. The test material was, therefore, tested up to the maximum recommended dose of 5000 µg/plate.

 

No significant increase in the frequency of revertant colonies was recorded for any of the bacterial strains with any dose of the test material, either with or without metabolic activation. The test material was considered to be non-mutagenic under the conditions of this test.