2-amino-6-methyl-4-propyl-1,2,4-triazolo[1,5-a]pyrimidin-5(4H)-one

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 Mar 1978 to 29 May 1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Kanppo Gyo No5 (Environmental Agency), Yakuhatsu No 615 (Ministry of Health and Welfare) and 49 Kiyoku No 392 (Ministry of International Trade and Industry)

Version / remarks:

Method for testing the degree of accumulation of chemical substances in fish body.

GLP compliance:

no

Radiolabelling:

no

Details on sampling:

Extracts with chloroform from test water of both groups were prepared twice (10 minutes shaking). The chloroform fractions were dehydrated (IPS filter paper), condensed by a rotary evaporator and made up to mL with chloroform before HLC analysis.

Vehicle:

no

Details on preparation of test solutions, spiked fish food or sediment:

2000 ppm (w/v) standard solution is prepared by dissolving 2g of the test compound in water and making it up to 1 L.

Test organisms (species):

other: carp (not further specified)

Details on test organisms:

TEST ORGANISM
- Weight at study initiation: 29 g
- Length at study initiation: 11 cm
- External sterilising: Fish are kept in steriliser bath (10 ppm solution of chlorotetracycline - HCl) for 24 hours
- Acclimation: At 25 °C for 14 days

Route of exposure:

aqueous

Justification for method:

other: not specified

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

8 wk

Test temperature:

23 - 27 °C

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 L aquarium
- Type: Flow through
- Inflow: 576 L/day

Nominal and measured concentrations:

- Group 1: 0.2 ppm (w/v)
- Group 2: 0.02 ppm (w/v)
See 'Any other information on materials and methods incl. tables' for the analysed concentrations during the test.

Reference substance (positive control):

no

Details on estimation of bioconcentration:

TREATMENT OF THE SAMPLE
At exposure termination, fish were weighed, slit into pieces and homogenised with the aid of 15 g of sea sand by addition of 100 g of anhydrous Sodium Sulphate. This was dehydrated, extracted with150 mL of chloroform (10 min shaken) and filtered by a glass filter G 4 (twice). The chloroform fractions were then dehydrated (1 ps filter paper) and condensed by a rotary evaporator. 10 mL of benzene was added and the solution was purified by column chromatograph (see HLC conditions below). The fractions were then condensed by a rotary evaporator and made up to 10 mL with chloroform.

HLC SPECIFICATIONS
- Filler: 10g of 3% hydrated Alumina (I C N Co Ltd) with benzene
- Column: 20 mm diameter glass tube
- Eluent: Fraction 1: 100 mL benzene; Fraction 2: 100 mL 20% chloroform/benzene; Fraction 3: 50% chloroform/benzene
The test item elutes to fraction 3.

Conc. / dose:

0.2 other: ppm

Temp.:

25 °C

Type:

BCF

Value:

>= 0.2 - <= 1.6 dimensionless

Basis:

not specified

Time of plateau:

8 wk

Calculation basis:

kinetic, corrected for growth

Remarks on result:

other: See 'Any other information on results incl. tables'

Conc. / dose:

0.02 other: ppm

Temp.:

25 °C

Type:

BCF

Value:

5.5 dimensionless

Basis:

not specified

Time of plateau:

8 wk

Calculation basis:

kinetic, corrected for growth

Remarks on result:

other: See 'Any other information on results incl. tables'

Details on results:

Results on the degree of accumulation is given in 'Any other information on results incl tables'. The highest accuracy is expected for analysis when the concentration of the test item is around 1.6 ppm. In the case that a fish has 30 g body weight, 10 mL of final extract is used for analysis and the recovery rate is estimated as 80 %, the degree of accumulation can be recalculated as follows in the fish body:

1.6 ppm/0.8*30/10 = 0.67 ppm

Equally the analytical limit of the test item, 0.1 ppm (signal/Noise = 2 : Noise = 1 mm) accounts for 0.04 ppm as the concentration in fish body. Therefore, the degree of accumulation of the fish with the test item level of 0.04 - 0.67 ppm is indicated as a reference in the parathesis and with the level of less than 0.04 ppm, it is indicated as <0.2 or >2.

RECOVERY
- Recovery blanks: 81.3 - 98.8 %
- Recovery group 1: 95.4%
- Recovery group 2: 81.5%

Table: Degree of bioaccumulation/ Accumulation factor (L)

	2 Weeks	4 Weeks	6 Weeks	8 Weeks
Group 1	(0.6)	0.2	(0.5)	(1.1)
	(0.6)	(0.3)	(0.8)	(1.6)
Group 2	< 2	< 2	< 2	< 2
	< 2	< 2	< 2	(5.5)

The degree of accumulation of the fish with the test item level of 0.04 - 0.67 ppm is indicated as a reference in the parathesis and with the level of less than 0.04 ppm, it is indicated as <0.2 or >2.

Table: Details on fish analysis results – 1

Sample	A - Injetion volume	B - Σ height (mm)	D - Apparent Conc. On chart (pp,)	E - Dilution factor	F - Final volume (mL)	G - Absolute amount in fish (µg)	H - Fish weight (g)	I - Recovery (%)	J - Conc. in fish (ppm)	K - Conc. In aquarium (ppm)	L - Accumulation factor	Injection fish
												M - Conc. (ppm)	N - Volume (mL)
												100	0.3
Standard 2.5 ppm	60	92.6										(30 µg)
Fish recovery group 2a	60	73.8	1.99	1.0	10.0	19.9		66.4
Fish recovery group 2b	60	66.7	1.80	1.0	10.0	18.0		60.0
							Ava.	63.2
Standard 2.5 ppm	60	93.2
Fish blank a	60	nd	-	1.0	10.0	-
Fish blank b	60	nd	-	1.0	10.0	-

 

Table: Details on fish analysis results – 2 

Sample	A - Injetion volume	B - Σ hight (mm)	D - Apparent Conc. On chart (pp,)	E - Dilution factor	F - Final volume (mL)	G - Absolute amount in fish (µg)	H - Fish weight (g)	I - Recovery (%)	J - Conc. in fish (ppm)	K - Conc. In aquarium (ppm)	L - Accumulation factor	Injection fish
												M - Conc. (ppm)	N - Volume (mL)
Standard 2.5 ppm	60	96.4
Fish 2 week group 1	60	7.2	0.187	1	10	1.87	25.4	63.2	0.116	0.184	0.6
Fish 2 week group 1	60	8	0.207	1	10	2.07	31.3	63.2	0.105	0.184	0.6
Standard 2.5 ppm	60	94.7
Fish 4 week group 1	60	2.4	0.064	1	10	0.634	27.5	63.2	0.036	0.186	0.2
Fish 4 week group 1	60	4.3	0.114	1	10	1.14	28.7	63.2	0.063	0.186	0.3
Standard 2.5 ppm	60	94.6
Fish 6 week group 1	60	6.8	0.18	1	10	1.8	29.2	63.2	0.097	0.18	0.5
Fish 6 week group 1	60	9.3	0.246	1	10	2.46	27.4	63.2	0.142	0.18	0.8
Standard 2.5 ppm	60	77.2
Fish 8 week group 1	60	11.7		1	10	3.79	30.2	63.2	0.199	0.179	1.1
Fish 8 week group 1	60	17.6		1	10	5.7	31.4	63.2	0.287	0.179	1.6

 

Table: Details on fish analysis results – 3 

Sample	A - Injetion volume	B - Σ hight (mm)	D - Apparent Conc. On chart (pp,)	E - Dilution factor	F - Final volume (mL)	G - Absolute amount in fish (µg)	H - Fish weight (g)	I - Recovery (%)	J - Conc. in fish (ppm)	K - Conc. In aquarium (ppm)	L - Accumulation factor	Injection fish
												M - Conc. (ppm)	N - Volume (mL)
Standard 2.5 ppm	60	95.9
Fish 2 week group 2a	60	nd	-	1	10	-	27	63.2	-	0.024	-
Fish 2 week group 2b	60	nd	-	1	10	-	27.2	63.2	-	0.024	-
Standard 2.5 ppm	60	95.3
Fish 4 week group 2a	60	nd	-	1	10	-	28.5	63.2	-	0.024	-
Fish 4 week group 2b	60	nd	-	1	10	-	30.1	63.2	-	0.024	-
Standard 2.5 ppm	60	93.2
Fish 6 week group 2a	60	nd	-	1	10	-	33.2	63.2	-	0.023	-
Fish 6 week group 2b	60	nd	-	1	10	-	28	63.2	-	0.023	-
Standard 2.5 ppm	60	78.3
Fish 8 week group 2a	60	nd	-	1	10	-	23.2	63.2	-	0.023	-
Fish 8 week group 2b	60	7	0.223	1	10	2.23	28.1	63.2	0.126	0.023	5.5

 

Validity criteria fulfilled:

not specified

Conclusions:

At a concentration of 0.2 and 0.02 ppm, the bioaccumulation factor ranged between 0.2 - 1.6 and <2 - 5.5, respectively, over 8 weeks in carp.

Description of key information

BCF of 0.2 - 5.5, Japanese test guideline, ICI Japan Ltd.

Additional information

The bioaccumulation potential of the test material was investigated in fish in a non-GLP study according to Japanese guidelines Kanppo Gyo No5 (Environmental Agency), Yakuhatsu No 615 (Ministry of Health and Welfare) and 49 Kiyoku No 392 (Ministry of International Trade and Industry) (Chemicals Inspection & Testing Institute, Japan, 1978). Two groups of carp (species unknown) were exposed to concentrations of 0.2 and 0.02 ppm to the test material for 8 weeks in a flow-through aquarium with an inflow of 576 L/day. After 2, 4, 6 and 8 weeks of exposure, analytical verification of the test material in water and fish was performed using HPLC analysis. The recovery of the test material was determined to be ≥80%. Based on the results, it was calculated that at a concentration of 0.2 and 0.02 ppm, the bioaccumulation factor ranged between 0.2 - 1.6 and <2 - 5.5, respectively, over 8 weeks in carp.

The two main impurities of the substance have predicted log Kow values of 2.31 and 2.48 (calculated with ACD/Labs, values as given in SciFinder), and the bioconcentration factors calculated with the bioaccumulation tool in US EPA Episuite 4 (BCFBAF v3.01) are 8.75 to 14.7 L/kg and 17.1 to 21.5 L/kg). These values show that also the impurities of the substance are unlikely to accumulate in biota to a significant extent.