Reaction mass of Resin acids and Rosin acids, hydrogenated, potassium salts and potassium [1R-(1α,4aβ,10aα)]-1,2,3,4,4a,9,10,10a-octahydro-7-isopropyl-1,4a-dimethylphenanthren-1-carboxylate

Bioaccumulation: aquatic / sediment

Administrative data

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Published and peer reviewed study performed according to sound scientific methods, but with slight differences from OECD guideline method.

Data source

Reference

Reference Type:

publication

Title:

FREE AND CONJUGATED CONCENTRATIONS OF NINE RESIN ACIDS IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) FOLLOWING WATERBORNE EXPOSURE

Author:

Niimi AJ and Lee HB

Year:

1992

Bibliographic source:

Environmental Toxicology and Chemistry, Vol. 11, pp. 1403-1407, 1992. Printed in the USA. Pergamon Press Ltd.

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The concentration of rosin acids were monitored in water in a flow through system and in exposed fish over a 30 day period. The fish were exposed to the resin acids over the first 20 days to estimate the BCF, followed by a 10 day depuration period to estimate elimination rate.

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

palustric acid

IUPAC Name:

palustric acid

Constituent 2

Reference substance name:

Chlorodehydroabietic acid containing equal amounts of 12- and 14-chloro isomers

IUPAC Name:

Chlorodehydroabietic acid containing equal amounts of 12- and 14-chloro isomers

Constituent 3

Reference substance name:

isopimaric acid

IUPAC Name:

isopimaric acid

Constituent 4

Reference substance name:

sandaracopimaric acid

IUPAC Name:

sandaracopimaric acid

Constituent 5

Reference substance name:

Abietic acid

EC Number:

208-178-3

EC Name:

Abietic acid

Cas Number:

514-10-3

IUPAC Name:

abieta-7,13-dien-18-oic acid

Constituent 6

Reference substance name:

pimaric acid

IUPAC Name:

pimaric acid

Constituent 7

Reference substance name:

Dichlorodehydroabietic acid

IUPAC Name:

Dichlorodehydroabietic acid

Constituent 8

Reference substance name:

Dehydroabietic acid

IUPAC Name:

Dehydroabietic acid

Constituent 9

Reference substance name:

neoabietic acid

IUPAC Name:

neoabietic acid

Details on test material:

The chemicals were purchased from Helix Biotech Corporation, Richmond, British Columbia. Purities reported for dehydroabietic acid, neoabietic acid, and isopimaric acid exceeded 98%; 95 to 99% for dichlorodehydroabietic acid; 90 to 95% for abietic acid and palustric acid; 85 to 95% for chlorodehydroabietic acid; and 85 to 90% for sandaracopimaric acid and pimaric acid.

Radiolabelling:

no

Sampling and analysis

Details on sampling:

Five trout were sampled just before dosing began and served as control fish. Five fish each were sampled again after 5, 10, 14, and 20 d exposure to estimate BCF. Only acetone was added to the tank after day 20, and five fish each were sampled after 24, 27, and 30 d to estimate the elimination rates of free and conjugated acids. Duplicate 1-L water samples were taken at 4- to 6-d intervals during the first 20 d and on day 24. Water samples were analyzed the same day.

Test solutions

Vehicle:

yes

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

A stock solution containing the acids dissolved in acetone was continuously added to the 350-L tank at 0.5 ml/min using a peristaltic pump (Gilson Medical Electronics Inc., Middleton, WI). This solution was diluted with a 3-L/min inflow of water. Mean waterborne concentration of each acid was between 0.7 to 3.6 ug/L (Table 1).

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The hatchery-reared trout, which averaged 140 g each, were held at 15 ± 1°C. Fish were fed every second day on a dry diet at about 1% of body weight.

Study design

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

20 d

Total depuration duration:

10 d

Test conditions

Hardness:

135 mg/L as CaC03

Test temperature:

15 ± 1°C

pH:

8.0 to 8.3

Details on test conditions:

Free acid concentrations were monitored in rainbow trout (Oncorhynchus mykiss) over a 30-d period. Fish were exposed to waterborne acids for the first 20 d to estimate BCF, followed by a 10-d acid-free period to estimate the elimination rate. A stock solution containing the acids dissolved in acetone was continuously added to the 350-L tank at 0.5 ml/min using a peristaltic pump. This solution was diluted with a 3-L/min inflow of water. Mean waterborne concentration of each acid was between 0.7 to 3.6 ug/L (Table 1). Five trout were sampled just before dosing began and served as control fish. Five fish each were sampled again after 5, 10, 14, and 20 d exposure to estimate BCF. Only acetone was added to the tank after day 20, and five fish each were sampled after 24, 27, and 30 d to estimate the elimination rates of free and conjugated acids. Duplicate 1-L water samples were taken at 4- to 6-d intervals during the first 20 d and on day 24.

Nominal and measured concentrations:

See Table 1

Reference substance (positive control):

no

Details on estimation of bioconcentration:

The means ± SDs of five fish were reported for free and conjugated acid concentrations at each sample interval unless noted otherwise. BCF was estimated by dividing the free acid concentration in each fish by the mean waterborne concentration. Analysis of variance (ANOVA) was used to test for differences in BCF among the 5- to 20-d sample intervals. Elimination rates were based on a first-order equation to estimate chemical half-life.

Results and discussion

Bioaccumulation factor

Remarks on result:

other: See Table 2

Details on results:

Fish exposed to 24ug/L total waterborne free acids retained about 2 mg/kg free and 1 mg/kg conjugated acids during the study (Table 1). There were no detectable levels of free or conjugated acids in fish sampled 4 to 10 d after acid exposure stopped, which prevented their half-lives from being calculated. Nevertheless, based on the <25- to 330-ug/kg concentrations of free and <25to 490 ug/kg concentrations of conjugate acids after 20 days of exposure, and the quantitation limit of 25 ug/kg after 4 d post-exposure, it could be suggested that the half-life for these free and conjugated resin acids would be <4 days in trout. BCF values for the eight detected free acids were highest after 10 d and decreased significantly (P≤ 0.05) thereafter (Table 2). A mean BCF for each free acid was estimated from the 14- and 20-d values. Mean BCF values ranged from about 25 for chlorodehydroabietic acid to 130 for neoabietic acid. The 5-day BCF values for some acids were not significantly different from those at 20 day, but they were not used to estimate mean BCF due to differences at the 10-day sample interval.

Any other information on results incl. tables

Table 1: Free and conjugated resin acid concentrations in trout exposed to waterborne concentrations of

0.7 to 3.6 ug/L after 5 to 20 days

Acid	Water Conc. (ug/L)^a		Conc. in fish (ug/kg)^b
		Form	5 d	10 d	14 d	20 d
Palustric	1.1 ± 0.7	Free	<25^c	<25^c	<25^c	<25^c
		Conj.	<25^c	<25^c	<25^c	<25^c
14 Chlorodehydroabietic	2.2 ± 0.6	Free	141 ± 33	127 ± 25	57 ± 18	44 ± 19
		Conj.	<28^d	146 ± 27	62 ± 32	109 ± 26
12 Chlorodehydroabietic	2.6 ± 0.5	Free	138 ± 21	122 ± 42	70 ± 26	59 ± 30
		Conj.	78 ± 22	102 ± 37	95 ± 47	186 ± 48
Isopimaric	2.7 ± 0.8	Free	122 ± 29	162 ± 19	91 ± 32	94 ± 46
		Conj.	136 ± 33	153 ± 78	93 ± 46	195 ± 62
Sandaracopimaric	2.6 ± 0.9	Free	165 ± 19	183 ± 20	126 ± 34	118 ± 44
		Conj.	79 ± 22	151 ± 78	67 ± 37	188 ± 53
Abietic	2.1 ± 0.9	Free	241 ± 31	297 ± 45	144 ± 52	146 ± 64
		Conj.	75 ± 51	234 ± 177	41 ± 18	119 ± 12
Pimaric	2.8 ± 0.8	Free	241 ± 31	298 ± 23	205 ± 58	197 ± 72
		Conj.	134 ± 21	223 ± 111	115 ± 36	262 ± 65
Dichlorodehydroabietic	3.2 ± 0.6	Free	380 ± 107	616 ± 78	300 ± 53	292 ± 130
		Conj.	299 ± 140	480 ± 174	253 ± 81	492 ± 119
Dehydroabietic	3.6 ± 1.5	Free	654 ± 146	650 ± 222	358 ± 126	333 ± 137
		Conj.	144 ± 75	63 ± 32	85 ± 48	79 ± 21
Neoabietic	0.7 ± 0.2	Free	98 ± 46	154 ± 20	86 ± 47	96 ± 43
		Conj.	<25^c	<25^c	<25^c	<40^e
Total	24	Free	2,200	2,630	1,460	1,400
		Conj.	1,020	1,600	860	1,690

^aMean ± SD of four samples.

^bMean ± SD of five samples, or mean when noted otherwise.

^cAll samples below quantitation limit of 25 ug/kg.

^dFour of five samples below 25 ug/kg.

^eThree of five samples below 25 ug/kg.

Table 2. Bioconcentration factor (BCF) of free resin acids in trout exposed to 0.7 to 3.6 ug/L

waterborne acids after 5 to 20 d

Acid	BCF^a
	5 d	10 d	14 d	20 d	Mean BCF^b
Palustric	<25	<25	<25	<25	<25
14 Chlorodehydroabietic	64 ± 15*	58 ± 11 *	26 ± 8	19 ± 10	23 ± 9 (10)
12 Chlorodehydroabietic	53 ± 8*	47 ± 16*	27 ± 10	22 ± 12	25 ± 10 (10)
Isopimaric	45 ± 11	60 ± 7*	34 ± 12	35 ± 17	34 ± 14 (10)
Sandaracopimaric	64 ± 7	70 ± 8*	49 ± 13	45 ± 17	47 ± 14 (10)
Abietic	115 ± 14*	141 ± 21*	69 ± 25	70 ± 31	69 ± 26 (10)
Pimaric	87 ± 16	107 ± 8*	73 ± 20	70 ± 26	72 ± 22 (10)
Dichlorodehydroabietic	119 ± 34	193 ± 24*	94 ± 16	91 ± 40	92 ± 29 (10)
Dehydroabietic	182 ± 40*	181 ± 62*	100 ± 35	92 ± 38	96 ± 35 (10)
Neoabietic	141 ± 65	220 ± 29*	121 ± 72	137 ± 61	129 ± 63 (10)

^aMean ± SD BCF of five fish at each sample interval. BCF values with asterisks are significantly different (P ≤ 0.05) from those at 20 d.

^bMean BCFs were calculated from values that were not significantly different from those at 20 d, although values at 5 d were not included if those at 10 d were different. Sample size in parentheses.

Applicant's summary and conclusion

Validity criteria fulfilled:

not applicable

Conclusions:

The 20 day exposure period is 8 days shorter than the OECD 305 method but it appears that the concentrations of all of the resin acids did reach equilibrium. Equilibrium was observed on two sampling occasions at days 14 and 20 which is one less sampling period required by OECD 305. Analytical procedures appeared to be appropriate.

Executive summary:

Rainbow trout (Oncorhynchus mykiss) were exposed to nine resin acids that included abietic, dehydroabietic, chlorodehydroabietic, dichlorodehydroabietic, neoabietic, pimaric, isopimarie, sandaracopimaric, and palustric acids. Bioconcentration factors (BCFs) for these acids ranged from <25 to 130 among fish that were exposed to mean waterborne concentrations of 0.7 to 3.6 ug/L for 20 days. Total free and conjugated acid concentrations ranged from 1 to 3 mg/kg on a wholebody basis. Elimination rates were also monitored over a 10-day deparation period, although no detectable levels were observed. Based on initial chemical concentrations, it was concluded that the half-lives of these acids were < 4 days.