Lysine hydrochloride

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20/02/2008-27/03/2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed according to the OECD guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: “VI Acute Toxicity Study with Fish” in the “On Test Methods of New Chemical Substances” (Nov 21, 2003, Yakushoku No. 1121002, Heisei 15-11-13 Seikyoku No. 2, Kanhoki No. 031121002)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Lysine hydrochloride

EC Number:

211-519-9

EC Name:

Lysine hydrochloride

Cas Number:

657-27-2

Molecular formula:

C6H14N2O2.ClH

IUPAC Name:

lysine hydrochloride

Constituent 2

Reference substance name:

L-lysine hydrochloride

EC Number:

233-234-9

EC Name:

L-lysine hydrochloride

Cas Number:

10098-89-2

IUPAC Name:

L-lysine hydrochloride

Details on test material:

- Name of test material (as cited in study report): L-lysine hydrochloride
- Analytical purity: 100.1 %
- Lot/batch No.: 070039
- Expiration date of the lot/batch: April 25, 2008
- Storage condition of test material: Dark and cool place

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The sample for analysis was collected with no replicate from mid layer of each test vessel without agitation.

Test solutions

Vehicle:

no

Details on test solutions:

The test solution was prepared by weighing 0.4995 g of the test substance (0.5000 g in terms of active ingredient), washing it with dilution water into a vessel, further diluting with the dilution water to make final volume of 1 L, and thoroughly being agitated with a Teflon rod. The test solution was prepared for the first time at the initiation of the study, and thereafter, every time the dilution water was renewed (every 24h).

Test organisms

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Japanese killifish
- Strain: Oryzias latipes
- Source: Matsuoka Suisan (Shiga Prefecture)
- Average total length: 3.0 cm (2.7-3.1 cm), n = 20, (At the completion of exposure)
- Average body weight: 0.22 g (0.18-0.25 g), n = 20, (At the completion of exposure)


ACCLIMATION
- Acclimation period: 98 days (Nov. 26, 2007 - Mar. 03, 2008)
- Acclimation conditions (same as test or not): yes
- Type and amount of food: TetraMin (Tetra Japan). Approx. 2 % of the body weight of the fish/day. None was fed from 48 hours before exposure start.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): The mortality for the period of 7 days before the initiation of exposure was less than 5 %. Only fish apparently in good health was used for the study.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

A preliminary study was conducted by using two nominal concentration levels of 10 and 100 mg a.i. /L. The mortality at 96 h of exposure was 0 % and no toxic signs were observed even at the highest concentration of 100 mg a.i./L .

Test conditions

Test temperature:

23 ± 2˚C

pH:

7.6-7.8

Dissolved oxygen:

6.7-8.4 mg/L

Nominal and measured concentrations:

Nominal: 100 mg/L
Average measured: 103 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquarium
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: glass, 21 x 16 x 23 H cm, 5L
- Aeration: no (At least 60 % of air saturation value was maintained)
- Renewal rate of test solution (frequency/flow rate): Batch-wise renewal of the test solutions every 24 hour
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated water (Municipal water of Takarazuka-City, which was treated with activated charcoal to eliminate any residual chlorine and adequately aerated) was used as dilution water.
- for more details see the below attachment


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours dark.
- Light intensity:Indoor lighting


TEST CONCENTRATIONS

- Range finding study: yes
- Test concentrations: 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: The mortality at 96 hours of exposure was 0 % and no toxic signs were observed even at the highest concentration of 100 mg a.i./L. Based on the above results, it was decided to conduct the definitive study as a limit study using one concentration level of 100 mg a.i. /L.

Reference substance (positive control):

yes

Remarks:

copper sulfate (II) pentahydrate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 103 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 103 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 103 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 103 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

- Results with reference substance valid? yes
- LC50: 2.3 mg/L.
- Other: sensitivity confirmation study was conducted every 6 month or for every lot

Any other information on results incl. tables

Sublethal observations / clinical signs:

Cumulative Mortality (%) of Oryzias latipe sand Toxic Signs at Each Observation Time

Nominal conc. (mg a.i./L)	No. of organisms	Item	Duration of exposure
			3 hours	6 hours	24 hours	48 hours	72 hours	96 hours
Non-treated control	10	Cum. mortality (%) Toxic sign	0 NA:10	0 NA:10	0 NA:10	0 NA:10	0 NA:10	0 NA:10
100	10	Cum. mortality (%) Toxic sign	0 NA:10	0 NA:10	0 NA:10	0 NA:10	0 NA:10	0 NA:10

Note: NA: No abnormalities

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

96 h median lethal concentrations (LC50) of L-lysine hydrochloride for Japanese Killifish (Oryzias latipes) was >103 mg a.i./L (based on the average measured concentration).

Executive summary:

The acute toxicity of L-lysine hydrochloride to Oryzias latipes (1 replicate/concentration, 10 fish/concentration) was assessed in a semi-static test according to OECD 203. The nominal test concentration was 100 mg/L. Measured concentrations in freshly prepared and used (24h after the test solution was prepared) ranged between 99% and 103% of nominal.The LC50 was >103 mg/L, based on averaged measured concentrations.