I:\MEPC\53\2-12.doc For reasons of economy, this document is pr inted in a limited number. Delegates are kindly asked to bring their copies to mee tings and not to request additional copies. INTERNATIONAL MARITIME ORGANIZATION



### IMO 

# E

MARINE ENVIRONMENT PROTECTION COMMITTEE 53rd session Agenda item 2 MEPC 53/2/12 15 April 2005 Original: ENGLISH

HARMFUL AQUATIC ORGANI SMS IN BALLAST WATER

Application for approval of an Active Substance for Ballast Water Management

Submitted by Germany

SUMMARY
Executive summary: This document contains an applica tion for basic approval of an Active Substance under the Procedure fo r approval of Ballast Water Management Systems that make use of Active Substances (G9). Due to the bulkiness and technical complexity of the information the annex is submitted in English only.

Action to be taken: Paragraph 5

Related documents: BWM/CONF/36, MEPC 53/2/1

1 Regulation D-3.2 of the International Conve ntion for the Control and Management of Shipsí Ballast Water and Sediments (BWM C onvention) stipulates that Ballast Water Management systems (BWMS) that make use of Active Substances to comply with the Convention shall be approved by the Organi zation. During its fift y-second session, the MEPC developed a procedure for approva l that is submitted as MEPC 53/2/1.

2 The procedure for approval defines principle as pects to be documented by data or testing (MEPC 53/2/1, paragraph 4.2.1) and some basic pr inciples for risk ev aluation (MEPC 53/2/1, paragraph 5.3). According to paragraph 6.1 of document MEPC 53/2/1, the Technical Group has to evaluate whether this information is complete and of sufficient quality.

3 The receiving competent authority in Germany has verified the dossiers and believes them to satisfy the data requirements of the IM O Draft Procedure for approval of Ballast Water Management systems that make use of Active Substances (G9).

4 Germany therefore submits the applying manu facturerís summary of the dossier in the annex to this document to the Organization fo r evaluation according to the procedure as described in MEPC 53/2/1. The complete dossier will be made available to the experts of the Technical Group with the understand ing of confidential treatment.

Action requested of the Committee

5 The Committee is invited to consider the annex and take action as appropriate.

***

### MEPC 53/2/12

I:\MEPC\53\2-12.doc 

### ANNEX



## Background Information and Worst Case Scenario of Environmental Emission for Ballast Water Treatment

! It is estimated that ~ 3 to 10 billion tonnes of ballast water are carried around the world each year. ! It is estimated that > 7,000 diffe rent species are carried around in shipsí ballast water each day. ! Some of these organisms pose a direct risk to human health:

! Fish farming and recreational waters might contribute to this risk, if Ö

### MEPC 53/2/12 ANNEX Page 2

I:\MEPC\53\2-12.doc 

### Ö the Standard of D-2 is not met

Regulation D-2 Ballast Water Performance Standard ñ Ships conducting ballast water management shall discharge < 10 viable organisms / m3 > 50 μ; and < 10 viable organisms / ml < 50 μ and > 10 μ; and discharge of the indicator microbes shall not exceed the specified concentrations.

The indicator microbes, as a human health st andard, include, but are not be limited to: Toxicogenic Vibrio cholerae (O1 and O139) with < 1 colony forming unit (cfu) / 100 ml or < 1 cfu per 1 gram (wet we ight) zooplankton samples; Escherichia coli < 250 cfu / 100ml; Intestinal Enterococci <100 cfu / 100ml.

Since more than 90% of the world seaborne ca rgo is handled and ship ped via the 20 biggest ports Ö

### MEPC 53/2/12 ANNEX

Page 3

I:\MEPC\53\2-12.doc 

### Ö surrounding waters and port areas ofte n turned into anaerobic status

Based upon the environmental emission scenarios developed for anti-foulants following worst case scenario might help to define an acceptabl e risk in the frame of IMOís Ballast Water Management Convention.

Port Statistics Selected Port: Rotterdam Water Volume: 840,000,000 t Number of incoming ships: 29,450/a; 80/d Total Gross Tonnage: 489,222,000 GT/a; 1,323,074 GT/d Average amount of Ballas t Water: 144,876,600t/a; 396,922t/d

Shipsí Profile:

Ship Type / Pump capacity[t/h] Number of Ships Gross Tonnage [t] Amount of Ballast Water [t] Pump out time [h]

Annual Daily Annual Daily Annual Daily Annual Daily 

### Bulker (4x1,000t/h) 8,000 22 170,000,000 465,753 51,000,000 139,726 12,750 35



### General Cargo (2x600t/h) 20,000 55 300,000,000 821,918 90,000,000 246,575 75,000 205



### Other (2x200t/h) 1,000 3 3,000,000 8,219 900,000 2,466 2,250 6

Total 29,000 80 473,000,000 1,295,890 141,900,000 388,767 90,000 246

### MEPC 53/2/12 ANNEX Page 4

I:\MEPC\53\2-12.doc 

### Ballast Water Treatment ñ Worst Case Scenario for Hydrogen Peroxide:

Initial Dose: 21 mg/l Retention (Degradation) time [h]: zero Predicted Environmental Concentration: 0.0099 mg/l Predicted No Effect Concentration: 0.0100 mg/l PEC/PNEC Ratio: 0.99

Factors not considered in above scenario: Degradation occurring during pump out time 10 d Tidal Period (exchange of total Water Volume in Port): 12.41 h Degradation during shipís voyage: 1 ñ 28 d

The Port of Rotterdam was chosen because of the detailed statistics available via:
http://www.portofrotterdam.com
If smaller ports are chosen for different scenario s number and size of ships have to be amended accordingly.

Conclusion: Since the natural concentrati on of Hydrogen peroxide in marine waters can reach up to 0.058 mg/l the PNEC of 0.01 mg/l seems to be very conservative.

During a field trial carried out in Kiel harbou r in November 2002, the concentration of Hydrogen peroxide in 20 t of Ballast Water treated with Peraclean Ocean, decreased from 17.2 mg/l to 1.4 mg/l within 24 hours as shown in following graph:

Abbau des Desinfektionsmittels in den Lagertanks Messung am 28.11.2002

6,7 6,8 7 7, 2 7,2 6, 9 17,2

15,1

13,8

9,6

8,1

1,4 9,9

6,7

5,9

4,8 4,3

0 0 5 10 15 20

Zeit nach der Behandlun g pH -W ert [-] H2O2-concentration [ m Peracetic acid [mg/l] permitted lower pH -value [-]

permitted upper H2O2-concentr a [ m g /l ]

### MEPC 53/2/12 ANNEX

Page 5

I:\MEPC\53\2-12.doc Technical Summary of Dossiers by the applying manufacturer for the evaluation of an active substance according to the Draft Procedure for Approval of Ballast Water Management Systems that make use of Active Substances

Introduction

The applicant is proposing a chemical ballast water treatment with Peraclean Æ Ocean using a special technology. Peraclean Æ Ocean is a proprietary formulation based on peroxygen chemistry. It has been demonstrated to effectively kill bacteria, yeas ts, viruses, molds, spores, algae, protozoa, zooplankton, phytoplankton, larval stages and fish eg gs in ballast water. It is most effective in combination with solid separation. Due to its unspecific mode of action as an oxidiser it is effective and no resistances have been observed so far. It is active at high concentrations of sediment and/or organic matter at pH values between 5 and 9 and in the temperature range of -5 to more than 40°C. It is a liquid that can easily be applied automatically wi th a specific proprietary technology.

The formulation has a long shelf life (> 95% activi ty after 1 year of stor age at 20°C). The product is effective at relatively low concentrati ons. The active ingredients of Peraclean Æ Ocean are not bioaccumulative or persistent, will ultimately degr ade under environmental conditions to oxygen, water and carbon dioxide and will not form any persistent metabolites. In the following assessment the headings are grouped according to document MEPC 53/2/1 of October 29, 2004. The original numbers of that document are given in brackets.

Identification (4.1)

Peraclean Æ Ocean is a proprietary formulation based on peroxygen chemistry. The main components

are:

15% PAA 14.3% H 2 O 2 26.5% acetic acid

• Hydrogen peroxide (H 2 O 2 ) is commonly viewed as an ìenvironmentally friendlyì or ìgreenì chemical. This is particularly due to the physi cochemical properties of this small inorganic water-soluble molecule. H 2 O 2 is non-persistent, has innocuous or even beneficial degradation products, and does not accumulate.

• Hydrogen peroxide is produced naturally by m any living organisms either within the organism itself or in the surrounding medium. For example blue-green algae produce H 2 O 2 under light irradiation at a rate of 1-50 μg/l per hour (Zepp et al , 1987).

• Hydrogen peroxide levels in natural waters range from 0.14 to 58 μg/l in seawater and from 0.34 to 109 μg/l in fresh water (ECETOC, 1993).

1 Data on effects on aquatic organisms (4.2.1.1; 5.3.9)

1.1 Acute aquatic toxicity

The existing data for marine organisms are summarized in table 1.

### MEPC 53/2/12 ANNEX Page 6

I:\MEPC\53\2-12.doc Table 1: Acute toxicity for marine organisms

Species Endpoint Result as PAA Result calculated as Peraclean Æ
Ocean Reference

Fish

Pleuronectes platessa 96 h LC 50 11 mg/L 73 mg/L Tinsley and Sims 1987a

Juvenile Macomona liliana 96 h LC 50 4.35 mg/L 29 mg/L Degussa, 2004b

Invertebrates

Crangon crangon 96 h EC 50 15 mg/L 100 mg/L Tinsley and Sims 1987b

Embryoniclarval stages

Mytilus edulis embryo 48 h EC 50

NOEC 0.27 mg/ml

0.13 mg/L 1.8 mg/ml

0.87 mg/ml Fairhurst 1987

Crassostrea gigas embryo 48 h EC 50

NOEC 0.28 mg/L

0.13 mg/L 1.87 mg/L

0.87 mg/L Butler 1987

Mytilus galloprovincialis larval development (age < 4 h post fertilisation) 48 h EC 50 NOEC survival EC 50 NOEC normal development EC 50 NOEC normal surviving 0.85 mg/L 0.7 mg/L

0.55 mg/L 0.35 mg/L

0.54 mg/L 0.35 mg/L 5.66 mg/L 4.69 mg/L

3.68 mg/L 2.34 mg/L

3.64 mg/L 2.34 mg/L Degussa, 2005a

### MEPC 53/2/12 ANNEX

Page 7

I:\MEPC\53\2-12.doc 

### Acute toxicity data for freshwater or ganisms are summarized in table 2.

Table 2: Acute toxicity to freshwater organisms

Species Endpoint Result as PAA Result calculated

as Peraclean Æ
Ocean Reference

Fish

Oncorhynchus mykiss 96 h LC 50 1.6 mg/L 10.6 mg/L Gardener and Bucksath, 1996

Lepomis macrochirus 96 h LC 50 3.3 mg/L 22 mg/L Terell 1987b/ 22 mg product/l Salmo gairdneri 96 h LC 50 13 mg/L 87 mg/L CEFIC PAR, 1986, Douglas and Pell, 1986a Invertebrates

Daphnia magna 48 h EC 50 0.5 mg/L 3.3 mg/L Douglas and Pell 1986b Paracalliope fluviatilis 96 h LC 50 2.14 mg/L 14.2 mg/L Degussa, 2004b

Fish

In the study of Tinsley and Sims (1987b), plaice ( Pleuronectes platessa ), a saltwater fish species, with a mean weight of 8.5 g was used. Test solutions were renewed daily during this semi-static test. The relatively high LC 50 could be due to the rapid degradation of PAA in seawater. The half-life in seawater is less than 1 hour and therefore the exposure during the fish test could have been low. In the test with juvenile shellfish ( Macomona liliana , saltwater species) a static protocol was used. The test was conducted in natural water with a low organic carbon content. The values are based on nominal concentrations. (Degussa , 2004b). In the study with rainbow trout ( Oncorhynchus mykiss cold-water species, freshwater) were conducted at temperatures of 12-14°C. A nalytical measurements of H 2 O 2 in the test solutions were performed. Between the start (0-hour) and the end of the test (96-hour) the decrease in the concentration of H 2 O 2 ranged between 14 and 26% (Gardener and Bucksath, 1996). In the study the warm-water bluegill sunfish ( Lepomis macrochirus ) have the test solutions were not renewed and the values are based on nomi nal concentrations (Terell, 1987b). In the CEFIC PAR, 1986 study and the study of Douglas and Pell, 1986a with Rainbow trout ( Salmo gairdneri, freshwater) a semistatic protocol according to OECD 203 was used. The values are based on nominal concentrations.

### MEPC 53/2/12 ANNEX Page 8

I:\MEPC\53\2-12.doc Invertebrates

The study with equilibrium PAA (12%) in brown shrimp (Crangon crangon ) revealed a 96-h LC 50 value of 15 mg PAA/l. The medium was renewed daily during the study but analytical concentration measurements were not made. The mean weight of these saltwater crustaceans was 1.25 g (Tinsley and Sims, 1987a). The larger size of the organism compar ed to daphnids may explain the lower sensitivity of the shrimp. The high LC 50 value could also be related to the rapid degradation of PAA in seawater The toxicity tests with diluted equilibrium PAA (15%PAA, 14 % H 2 O 2 , 28% acetic acid) in the fresh water flea Daphnia magna was conducted using a static pr otocol according to OECD 202 part 1, without analysis of the PAA, active oxygen or H 2 O 2 concentrations in the test solutions (Douglas and Pell, 1986b). The LC 50 was 0.5 mg/L PAA. A static study with the freshwater amphipod Paracalliope fluviatilis revealed a 96 h LC 50 of 2.14 mg/L based on nominal concentrations.
Embryonic-larval stages

Two embryo-larval assays were carried out on marine oysters with nominal concentrations of equilibrium PAA (12.5%) dissolved in seawater, to determine any effects during their first 48 hours of their development from embryo to larva. In this period a protective D-shaped shell was formed. Without a shell the embryos were extremely sensitive to reduc ed water quality. Both tests were conducted under static conditions. In the first test with embryos of the pacific oyster ( Crassostrea gigas ), a 48-hour EC 50 of 0.28 mg PAA/l was obtained (Butler, 1987). Another test with embryos of the common mussel ( Mytilus edulis ) resulted in an EC50 value of 0.27 mg PAA/ l (Fairhurst, 1987). A recent study with Peraclean Æ Ocean according to US-EPA guideline EPA/ 600/R-95/136, 1995 re vealed a 48-hour EC 50 value for normal surviving larvae of 0.54 mg PAA/L (or 3.64 mg Peraclean Æ Ocean/L) and a NOEC of 0.35 mg PAA/L (2.34 mg Peraclean Æ Ocean/L). This was the most sensitive test in a number of screening tests with marine organisms tested in this study (see also 1.3) (Degussa, 2005a).

Conclusion

All fish toxicity data were of a similar order of magnitude and there seems to be no big difference between marine and freshwater species, if any marine species se em to be slightly less sensitive, but this could also be related to the more rapid degradation of PAA in salt water. Based on the most conservative value the acute LC 50 for fish is 1.6 mg/L.

In invertebrates the lowest EC 50 was obtained for Daphnia magna, with 0.5 mg PAA/L. The toxicity seems to be related to the size of the test organism. Embryonic stages were most sensitive with 48 h EC 50 values of 0.27 to 0.54 mg PAA/L.
Acute toxicity to bacteria

A luminescent bacteria test according to DIN EN ISO 348 Part 2 was performed with a comparable material. The concentration leading to below 20% inhi bition in 30 minutes was 0.83 to 1.25 mg product/l (corresponding to 0.125 to 0. 18 mg PAA/l) (Degussa, 2001a). In a respiration inhibition test according to OECD 209 and GLP) dilutions of Peracetic acid (39.5% PAA, 40.7% Acetic acid and 2.3 % hydrogen peroxide) were tested. After 30 minutes the EC 50 was 13.4 mg/L of the product corresponding to 5.3 mg/L of PAA, after 3 hours the EC 50 value was 12.9 mg/L product or 5.1 mg PAA/L (PAR, 2001).
For hydrogen peroxide a respiration inhibition te st according to OECD 200 and GLP revealed an EC 50 (3 h) of 466 mg/ml. EC 10 value for pseudomonas putida in an 18 h multiplication inhibition test was determined to be 11 mg/L (EU, 2003).

### MEPC 53/2/12 ANNEX

Page 9

I:\MEPC\53\2-12.doc 1.2 Chronic aquatic toxicity The available chronic toxicity data for the aquat ic compartment are summarized in table 3.

Table 3: Chronic aquatic toxicity data

Species Endpoint Result as PAA Result calculated as Peraclean Æ
Ocean Reference

Fish Menidia beryllina (larvae fish) 7-day survival and growth test LC 50 /IC 50 survival: 5.34 mg/L Biomass: 6.21 mg/L NOEC Survival : 1.88 mg/L Biomass: 3.75 mg/L LC 50 /IC 50 survival: 35.6 mg/L Biomass: 41.4 mg/L NOEC Survival : 12.5 mg/L Biomass: 25 mg/L Degussa 2005a

Invertebrates Daphnia magna 21-day reproduction test Parental immobilisation 21-day EC 50 : 0.1 mg/L Reproduction: 21-day EC 50 : 0.08 mg/L NOEC parental immobilisation and reproduction: 0.05 mg/L Parental immobilisation 21-day EC 50 : 0.61 mg/L Reproduction: 21-day EC 50 : 0.55 mg/L NOEC parental immobilisation and reproduction: 0.34 mg/L CEFIC, PAR, 2000

Mysidopsis bahaia 7-day survival and growth testing LC 50 /IC 50 survival: 2.52 mg/L Biomass: 2.70 mg/L NOEC Survival : 1.88 mg/L Biomass: 3.75 mg/L LC 50 /IC 50 survival: 16.8 mg/L Biomass: 18.0 mg/L NOEC Survival : 12.5 mg/L Biomass: 25 mg/L Degussa, 2005

Embryonic stages Marine blue mussel embryo Mytilus galloprovinc ialis embryo Development, 72 h NOEC: 0.3 mg/L LOEC: 0.59 mg/L EC 50 : 0.69 mg/L NOEC: 2 mg/L LOEC: 3.9 mg/L EC 50 : 4.6 mg/L Degussa 2004b

Algae Marine Minutocellus polymorphus Growth, 48 h NOEC: 0.59 mg/L LOEC: 1.19 mg/L EC 50 : 0.82 mg/L NOEC: 3.9 mg/L LOEC: 7.9 mg/L EC 50 : 1.64 mg/L Degussa, 2004b

Fresh water Pseudokirchneriella subcapita (Selenastrum capricornutum) Growth, 126 h NOEC: 0.13 mg/L EC 50 : 0.18 mg/L NOEC: 0.28 mg/L EC 50 : 0.4 mg/L Hicks et al

, 1996

Fresh water Pseudokirchneriella subcapita (Selenastrum capricornutum) Growth, 72 h NOEC: 0.074 mg/L LOEC: 0.15 mg/L EC 50 : 0.14 mg/L NOEC: 0.49 mg/L LOEC: 1 mg/L EC 50 : 0.93 mg/L Degussa, 2004b

### MEPC 53/2/12 ANNEX Page 10

I:\MEPC\53\2-12.doc A 21-day Daphnia magna reproduction study performed according to OECD guideline No. 211 and under GLP was performed using concentrations of 0.034, 0.11, 0.34, 1.1 and 3.4 mg/L of a comparable material. The concentration of PAA corresponded to 0.005, 0.016, 0.05, 0.16, 0.5 mg/L. T he test solutions were renewed 3 times per week. The NOEC of this study was 0.34 mg/L as Peraclean Æ Ocean or 0.05 mg PAA/L for both parental toxi city and reproduction. Two 7-day survival and growth assays were performed with a larval fish Menidia beryllnia and a mysid Mysidopsis bahia , both salt water organisms according to US EPA, 2002 guideline EPA-821-R-02-014 the test soluti on was renew ed daily. For M. beryllnia the LC 50 for survival corresponded to 5.34 mg PAA/L, for biomass to 6.21 mg PAA/L (35.6 and 41.4 mg Peraclean Æ Ocean/L respectively), the NOECs were 1.88 and 3.75 mg PAA/L (12.5 and 25 mg Peraclean Æ Ocean/L). For M. bahia the LC 50 /IC 50 for survival corresponded to 2.52 mg PAA/L, for biomass to 3.75 mg/L (16.8 and 18.0 mg Peraclean Æ Ocean/L respectively), the NOECs to 1.88 and 3.75 mg PAA/L (12.5 and 25 mg Peraclean Æ Ocean/L), (Degussa, 2005a). A 72 h static study with blue mussel embryo ( Mytilus galloprovincialis embryo) revealed the most sensitive marine species with an EC50 of 0.69 mg PAA/L and a NOEC of 0.3 mg/L. The toxicity to the marine algae (48 h) based on growth rate was slightly higher but of the same order of magnitude. Both values were based on nominal concentrations (Degussa, 2004b). A study with a freshwater green alga ( Selenastrum capricornutum ) and equilibrium PAA (5.2%) has been reported. The study was performed to good laboratory practice (GLP) guidelines. The test concentrations were 0, 0.065, 0.13, 0.25, 0.50 and 1.0 mg PAA/l. Growth was det ermined by algal cell counts. The concentration of H 2 O 2 was measured in the test solutions. At the start of the test, the H 2 O 2 content of all test solutions was close to the nominal concentration. At the end of the test, the measured concentration was lower than the detection limit except for concentr ations of 0.50 and 1.0 mg PAA/l where the remaining measured H 2 O 2 concentrations were 84% and 110% of the nominal values, respectively. At a concentration of 0.13 mg PAA/l an initial, statisticall y significant, inhibition of growth was found between 0 and 72 hours. Growth had recovered at the end of the test probably due to decreased exposure. Based on the cell count at the end of the test, a concentration of 0.13 mg/L was considered to be an NOEC. At a concentration of 0.25 mg/L the cell count at the end of the test was 3% of the control value. The 120-hour EC 50 value was estimated to be 0.18 mg/L (Hicks et al , 1996). Comparable results were obtained in a study with the same algae and 72-h duration by Degussa, 2004b.



### Conclusion

The results of the available chronic studies are remark ably similar and indicate an overall NOEC value of around 0.05 to 0.1 mg PAA/L. The studies all indicate a very steep dose response. The chronic toxicity does not differ much from the acute toxicity (factor 5) . Through the unspecific mode of action (oxidation) the toxicity depends very much on the size of the respective organism that may be related to its detoxification capacity. Based on th e availability on studies on very se nsitive embryonic states that are normally not available together with some long term to xicity data an assessment factor of 10 on the lowest NOEC of 0.05 mg PAA/L may be appropriate leading to a PNEC of 5 μg/L. Given the consistency of the data, the low difference between acute and chronic effects, the mechanism of action and the steep dose response, and the instability of the substance in wa ter a lower assessment factor of 5 may also be regarded as sufficient and a PNEC of 10 μg/L seems to be sufficiently conservative. This would correspond to a PNEC of 67 μg Peraclean Æ Ocean/L.



### 1.3 Aquatic toxicity data on hydrogen peroxide

The most important aquatic toxicity data of hydro gen peroxide are summarized below, because from the results of preliminary experiments with ballast wa ter treatment it can be concluded hydrogen peroxide may be released after treatment, while peracetic acid could not be detected anymore.

### MEPC 53/2/12 ANNEX Page 11

I:\MEPC\53\2-12.doc For hydrogen peroxide the data have been reviewed in EU 2003. Selected data from this report that were used for the risk characterisation are outlined in table 4.

Table 4: Aquatic toxicity data for hydrogen peroxide

Trophic level Species Endpoint Result Reference Fish Pimephales promelas 96 h LC 50 , semistatic, concentration measured 16.4 mg/L EU 2003

Aquatic invertebrates Daphnia pulex 48 h EC 50 , semistatic, concentration measured 2.4 mg/L EU 2003

Daphnia magna 24 h EC 50 , static nominal concentration 2.3 mg/L EU 2003

Gammarus sp. 96 h EC 50 , semistatic, nominal concentration 4.4 mg/L EU 2003

Physa sp. 96 h, semistatic, nominal 17.7 EU 2003

Dreissena polymorpha (zebra mussel) 56 day chronic study, flow through NOEC: 2 mg/L 56-d LC 30 : 5 mg/L EU 2003

Algae Chlorella vulgaris 72 h chronic study EC 50 (growth): 2.5 mg/L EC 50 (growth rate): 4.3 mg/L NOEC: 0.1 mg/L EU 2003

Sceletonema costatum (marine diatom) 72 h chronic study EC 50 : 1.38 mg/L NOEC: 0.63 mg/L EU 2003

Long-term studies are available for 2 trophic levels including the most sensitive (invertebrates). Furthermore some field studies are available indicati ng that ecosystems may be able to tolerate varying concentrations of hydrogen peroxide and are adapt ed to changing concentrations of hydrogen peroxide due to photochemical generation of the substance in wa ter systems. In seawater the limited amount of available natural concentrations ranged from 0.5 to 14 μg hydrogen peroxide/l, in lakes values > 100 μg/l were reported (EU, 2003). In EU, 2003 a PNEC of 10 μg/L for hydrogen peroxide was derived. However, this value seems to be very conservative and is re lated to continuous long term exposures. When deriving a tolerable level for hydrogen peroxide in balla st water applications the following considerations should be taken into account: The mechanism of action of hydrogen peroxide

H 2 O 2 acts as a strong oxidant. Such reactivity m eans that the toxicity effects will be the immediate consequence of oxygen, originated from the decomposition of H 2 O 2 by local reaction with biological tissues. Therefore it is expected that the adverse effects of hydrogen peroxide are only local and short term. The natural defence mechanisms against H 2 O 2 Most aerobic living organisms are particularly we ll equipped in defence mechanisms allowing them to inactivate hydrogen peroxide and other reactive oxygen species. Antioxidant enzymes and oxygen scavengers have been observed in invertebrates (e.g . crustacean, molluscs, annelida, plathelminthes, coelenterata, porifera), in vertebrat es (fish) such as catalase, superoxide dismutase, glutathione and glutathione peroxidase and carotenoids (Lemaire and Livingstone, 1993; Morris and Albright, 1981) as well as in algae (catalase and peroxidases), both highe r and less evolved. This protection mechanism arises in biological systems during evolution due to the continuous formation of reactive oxygen species

### MEPC 53/2/12 ANNEX Page 12

I:\MEPC\53\2-12.doc (ROS) as unwanted toxic by-products of metabolism. On this basis, it is expe cted that aerobic organisms are well protected against the effect of hydrogen peroxide exposed to organisms.

Natural sources of H 2 O 2 As mentioned above hydrogen peroxide is naturally prod uced by sunlight in water and is furthermore a common product of natural oxidative proces ses taking place in aerobic cells. H 2 O 2 may also be formed in the medium surrounding an organism where certain met abolites from the organisms act as promoters, e.g. riboflavin, excreted by a number of marine organisms (Moffet and Zika, 1987). The specific conditions in laboratory tests The medium used in laboratory tests is quite differ ent from the natural conditions. Depending on the environmental conditions, many materials (e.g. dissolved transition metals, solid metals, solid metal oxides, hydroxides, activated carbon, enzymes) contribu te to reduce the oxidant power. It is thus assumed that under laboratory conditions the test organisms are more exposed and thus more sensitive than under natural conditions to the prolonged exposure of H 2 O 2 concentrations during the test as illustrated by the dat a of Fitzpartrick et al , (1995) on fish. Also, in the field conditions, the more sensitive species are protected from a direct effect of hydrogen peroxide by the presence of the total biomass and the suspended solids. 

### 1.4 Endocrine disruption

From the structure of the active ingr edients, the fact that it is rapidly degraded and will not be available in higher organisms it can be concluded that Peraclean Æ Ocean is of no concern with regard to possible endocrine effects. 

### 1.5 Sediment toxicity

From the physical chemical proper ties, the high water solubility, the low log Kow and the instability in water it is unlikely that the components of Peraclean Æ Ocean will reach the sediment to a considerable extend. This is corroborated by Ma ckay calculations showing practica lly no distribution into sediment (see chapter 4). Accumulation in sediment is not to be expected. Sediment testing is therefore not considered necessary. 

### 1.6 Bioavailability/biomagnification/biocon centration/Food web population effects

Due to the low log Kow and the rapid degradation of the components of Peraclean Æ Ocean either by abiotic mechanisms or by all biota it is not likely t hat the product is bioavailable. Peracetic acid and hydrogen peroxide are rapidly degraded in the environm ent e.g. under the influence of transition metals and in biota by catalases and peroxidases that ar e ubiquitary in organisms (ECETOC 1993, 2001). Consequently no bioconcentration, biomagnification or accumulation along the food chain is to be expected. 2 Data on mammalian toxicity (4.2.1.2; 5.3.12)



### 2.1 Toxicokinetics

From the physical chemical properties absorption of Peraclean Æ Ocean through skin and mucous membranes is possible, but limited by the high wate r solubility and low partition coefficients of the equilibrium compounds. However, a considerable inta ke of radioactivity was observed in damaged skin, once the skin barrier is destroyed by the corrosive effects of PAA so lutions (Phillips, 1994a). Phillips studied the fate of 14C-labelled PAA (5.02%) afte r dermal application (100 μl to an area of 2.5 cm 2 ) to 4 male Sprague-Dawley rats. An acetate control treatment was included in the test. Water-soluble vapours (i.e. evaporating PAA and HOAc), exhaled CO 2 , urine and faeces were collected and analysed at regular intervals. After 72 hours the animals were k illed and the total radioactivity content was determined of the internal organs. When the recovered radioac tivity is presented as a percentage of the absorbed dose, significant differences between the test and cont rol animals were observed. In the acetate control animals 31% of the absorbed radioactivity was eliminat ed in urine versus 17% in the test group animals. In control animals 50% was exhaled as 14CO 2 versus 58% in the test group animals. Recovery in faeces was 4% in controls and 6 % in test group animals. Recovery in the tissues and carcass was 13% in controls and 20% in test group animals. The results of this study indicate that PAA is largely eliminated

### MEPC 53/2/12 ANNEX Page 13

I:\MEPC\53\2-12.doc within 72 hours of exposure with excretion of radioactivity occurri ng primarily via exhaled air (58 %), followed by urine (17 %) and faeces (6 %). It should be realised that the distribution of 14C is measured and that it is not known whether th is is present as PAA, HOAc or CO 2
. In a most recent study on the degradation of Peraceti c acid in rat blood (CEFIC PAR, 2004) it could be demonstrated that the half life of Peracetic acid in a 1000 fold diluted rat blood sample is below 5 minutes. Several authors have shown that peracetic acid is a substrate of catalases yielding acetic acid and oxygen (Ferri et al , 1990, Jones and Middlemiss, 1972, Placic a nd Dunford, 1980). PAA is also degraded by calf serum (M ̧cke, 1977), stomach fluid and saliva (Juhr et al , 1978). Hydrogen peroxide is also degraded rapidly by catalases, peroxidases and a num ber of other enzymes and antioxidants in the body (EU, 2003).
The importance of the catalase re action for the metabolism of PAA can be illustrated by looking at the distribution of catalases in the mammalian organism. Catalases are present at a wide range of concentrations in nearly all mammalian cells; the enzymes are particularly efficient in metabolising large amounts of H 2 O 2 (Chance et al , 1979). Catalases are located in sub-cellular compartments, mainly in peroxisomes (De Duve and Bauduin, 1966). Soluble catalases were found in erythrocytes (Saito et al , 1984).
The highest catalase content is observed in cells of the duodenum, liver, spleen, kidney, blood, mucous membranes and other highly vascularised tissues; the lowe st catalase activity occurs in brain, thyroid, testes and connective tissue cells (Matkovics and Novak, 1977). A more detailed discussion of catalase activity, inter- and intra-species differences can be found in ECETOC, 1993 and 1996.

Evaluation
The data on metabolism indicate that peracetic acid is rapidly degraded in various tissues and is expected to be degraded to a considerable extend at the site of first contact. Once absorbed into the circulation it is almost instantly degraded in the blood and will not be av ailable to other organs beyond the site of first contact.

Conclusion

The systemic absorption of peracet ic acid and hydrogen peroxide is limited by their rapid enzymatic degradation in cells of mucous membranes lining the gastr ointestinal and respiratory tract and to a certain extend also in skin. Tissue damage, for example on c ontact with severely irritant concentrations of Peracetic acid can lead to absorption into the circul ation. In blood peracetic acid and hydrogen peroxide are very rapidly degraded yielding oxygen and acetic acid or oxygen and water respectively. The remaining acetic acid is a physiological substance that is included into physiological metabolic pathways or excreted via the urine.

The rapid degradation limits the systemic availability of both peracetic acid and hydrogen peroxide. It is therefore unlikely that organs bey ond the site of first contact w ill be affected by peracetic acid.



### 2.2 Acute toxicity

The acute toxicity data of Peraclean Æ Ocean are summarized in table 5.

Table 5: Acute toxicity studies

Route of exposure LD 50 or LC 50 Species Reference Oral 1015 mg/kg bw Rat Degussa 1982a Dermal 1912 mg/kg bw Rabbit Kuhn 1996a Inhalation, aerosol Ca. 0.5-1.3 mg/L/4 h Rat Terrell, 1986

In the oral study, signs indicative of irritation of t he gastro-intestinal tract (writhing syndrome, stilted gait, and tremor), laboured breathing and bloody lachrymation were observed. Red coloured urine was also observed in females of the highest dose group. At necropsy adhesions were observed between the

### MEPC 53/2/12 ANNEX Page 14

I:\MEPC\53\2-12.doc viscera and the peritoneum. The gastro-intestinal mucosa and parts of the liver close to the stomach appeared white or greyish in animals of the high dose group. In the lowest dose group no clinical signs indicating irritation were observed. This dose co rresponded to a concentration of about 3% PAA, 4.5% H 2 O 2 and 4% HOAc. Symptoms observed in the dermal study included predominantly local effects on the skin as moderate to severe erythema, slight to severe oedema and eschar formation. General symptoms included decreased activity, diarrhoea, nasal discharge, ptosis, saliv ation and slow gazing. No pathological changes attributable to treatment were observed at necropsy. Terrel (1986) exposed rats to aerosol concentrations of PAA ranging from 18 to 2,138 mg/m 3 . The animals were kept under observation for 14 days af ter the exposure. During the exposure the animals showed a range of clinical signs which consisted of blinking, foaming, gasping, nasal discharge, salivation, lachrymation, ptosis, laboured respiration, twitching, chewing motion, convulsions, staggering, cornea opacity or death depending on the concentrati on. Post-exposure clinical signs were laboured respiration, nasal discharge, ga sping, lachrymation, haemorrhage fr om the eye, cornea opacity, blindness, staggering, loss of righting reflex, crusty appearance, piloerection or death depending on the dose. Gross necropsy of the animals that died during the observation period revealed alterations to the lungs and thymus, enteritis, swollen nose, conges ted nasal cavity and trachea, thickening of the oesophagus and larynx. Animals that were killed at t he end of the observation period showed similar alterations. Like all inhalation studies with perace tic acid this study suffered from the difficulty of generating and maintaining a stable atmosphere of the test item. An accurate measurement of the composition of the test atmosphere and particle size of the aerosol is almost impossible due to the instability of the product in air. The resulting LC 50 values should therefore be treated with circumspection.

Conclusion

The predominant effect in all acute toxicity studies is local irritation at the site of contact, the extent of which depends on the concentration and the co mposition of the applied test solution.



### 2.3 Effects on skin and eye

Skin irritation

Peraclean Æ Ocean was severely corrosive to rabbit skin and caused severe skin damage already after 3 min and 4 h of occlusive exposure. (Degussa 1982b, Degussa, 1990). Exposure of the skin of one animal to 0.5 ml PAA 15% for 3 minutes under occl usive conditions resulted in white to yellowish discoloration of the application site and deepening of the treated skin area. Severe erythema was observed at the border of the application site and sl ight oedema was reported. Microscopic examination revealed complete coagulation necrosis of the epi dermis and upper third of the corium including skin adnexae. The damage developed within 1 hour. It wa s concluded that 15% PAA was corrosive after 3 minutes of occlusive exposure (Degussa, 1990).

Skin sensitization

A skin sensitisation study using the B ̧hler method wa s conducted on short-haired albino guinea pigs to determine if a 5% PAA (20% H 2 O 2 and 10% HOAc) formulation could in duce dermal sensitisation (Kuhn, 1996b). The animals were treated (1 x /wk) with 0.4 ml of a 10% solution of the test compound in deionised water for 3 weeks. After a 2-wk rest period, the animals were challenged at a virgin test site with an application of 0.4 ml of a 7% solution of the test substance. After the challenge, a very faint erythema was observ ed in both the control a nd the treated group and, th erefore, 5% PAA was not considered to have elicited a sensitising reaction in guinea pigs. A similar study was conducted with 12% PAA (20% H 2 O 2 and 20% HOAc) (Kuhn, 1996c). In this case the animals were challenged with 0.4 ml of a 0.5% solution of the test substa nce in deionised water. The result s showed that 12% PAA did not elicit a sensitising reaction in guinea pigs.

Eye irritation

Cascieri and Freeman (1983) tested 0.1 ml of 17% PAA in rabbit ey es and found that it was extremely irritant to washed and unwashed eyes.

### MEPC 53/2/12 ANNEX Page 15

I:\MEPC\53\2-12.doc Respiratory tract irritation

Janssen (1989) nebulised a 15% PAA (14% H 2 O 2 , 28% HOAc) solution into an exposure chamber for 25 minutes. Male Wistar rats were exposed (nose-only) to the aerosol at concentrations ranging from 9.5 to 40.3 mg PAA/m 3 (3.7 to 14.3 mg H 2 O 2 /m 3 ; HOAc not measured) and their respiration rate was monitored. The RD50, referring to a concentration of PAA inducing a 50% reduct ion of respiratory rate, was calculated to be 21.5 to 24.1 mg/m 3 . Reduction of respiratory rate occurred at levels from 5 to 10 mg PAA/m 3 . After termination of exposure the respiratory rates returned to normal and the animals recovered fully within 3 days. No other clinical signs and no histopathological changes were observed.

Conclusion

Peraclean Æ Ocean should be considered severely corrosive to skin and eyes and severely irritating to mucous membranes of the respiratory tract. The RD 50 for an aerosol generated from a product with a similar composition was 21.5 to 24.1 mg/m 3

. No skin sensitisation was observed in two B ̧hler tests in guinea pigs wi th different formulations of PAA. These studies are considered relevant for Peraclean Æ Ocean as well as the study design requires testing with a dilution yielding the maximum non irritating concentration.



### 2.4 Repeated dose toxicity (Chronic and longterm toxicity)

A 90-day oral (gavage) (CEFIC, PAR 2003). 5.5% PAA containing 15.3% hydrogen peroxide and 16.6% acetic acid was used as the test substance. Groups of 5 male and 5 female Sprague Dawley rats received the following concentrations and dose levels by a daily gavage dose for 90 days.

Study days Group 1 Conc. [mg/L]/ dose mg/kg bw Group 2 Conc. [mg/L]/ dose mg/kg bw Group 2 Conc. [mg/L]/ dose mg/kg bw 1-10 10/15 33.3/50 100/150

11-22 10/15 33.3/50 66.7/100

23-92 3.33/5 10/15 33.3/50

The dose levels had to be adjusted several times due to mortality in the high dose group. One animal also died in the medium dose group when the dose level was 50 mg/kg bw. The main findings in the post-mortem observations of the animals that died duri ng the study were distended gastrointestinal tract due to gas formation, reddish coloured and or dilated lungs, reddish or blackish areas in the stomach mucosa. The findings indicate a massive gas (oxygen) formation in the gastrointestinal tract that could have led to asphyxiation of the stomach content s in the lungs and subsequent damage of lung and trachea cells as well as a considerable irritation at the site of contact. Clinical signs as ptyalism and loud breathing were consistent with the pathological findi ngs. No treatment related findings were reported in any other organs. No changes in haematological or c linical chemistry parameters were observed. No treatment related pathological findings (macroscopic and mi croscopic) were also observed in all surviving animals of all dose groups. The effects observed can therefore rather be attri buted to an acute irritant reaction following the gavage administration than to any sy stemic toxicity. The NOAEL of this study was 5 mg/kg bw because the dose level was reduced to this level in the low dose group at day 23 of 5% PAA, but it is likely that 15 mg/kg bw would also have been tolerated as no effects were observed in group 2 once the dose level was reduced to 15 mg/kg bw after day 23. The NOAEL would correspond to approximately 1.7 mg/kg bw per day or 5 mg/kg bw per day respectively of Peraclean Æ Ocean. However as the effects were related to local irritation only and no indications of systemic toxicity was observed, it is likely that they are more related to the concentration than to the dose and the NOAEC (no observed adverse effect concentration w ould correspond to 0.55 mg PAA/ 1.53 mg hydrogen peroxide/l and 1.66 g acetic acid/l.

### MEPC 53/2/12 ANNEX Page 16

I:\MEPC\53\2-12.doc Limited literature data are available for repeated dose st udies up to 10 moths duration in different species: rats, mice, Gebrils, guinea pigs, hamsters (e.g. Juhr et al , 1977, 1978). These have been reviewed and evaluated in ECETOC (2001). Alth ough these studies are of limited re liability they do not indicate any specific systemic toxicity of peracetic acid.

Conclusion

Repeated does studies with diluted equilibrium PAA solutions are relevant for Peraclean Æ Ocean as well as the test substance has to be diluted to tolerable concentrations before the administration to the test animals. No further repeated toxicity studies are considered necessary with Peraclean Æ Ocean for the following

reasons: The available studies indicate no specific syst emic toxicity of equilibrium peracetic acid. These findings are in accordance with the toxicokineti c data (see chapter 2.2.1) and support that peracetic acid will not be systemically avail able beyond the site of first contact.

The effects at the site of contact are determined by the acute irritation potent ial of the compound and its ability to release oxygen. It is not expected that this mode of action and the accompanying no effect levels will vary considerably between spec ies and with the duration of the study.

Therefore further subchronic and chronic studies with PAA are unlikely to result in new information and should not be performed based on animal welfare considerations.



### 2.5 Developmental and reproductive toxicity

As outlined in the Toxicokinetics section (3.1) it is very unlikely that PAA or hydrogen peroxide are systemically available and could reach the sex organs or the developing embryo or foetus. This is due to the rapid metabolism of PAA and hydrogen peroxide at the site of first contact and in the blood. This is also corroborated by the recent 90 day study in rats (CEFIC, PAR 2003) that did not show any effects on the testes or ovaries. A limited number of literature data, that are however, poorly reported are also available on this endpoint and were reviewed in ECETOC , 2003. In a teratogenicity study with ICR mice, the animals (5-10/group) were exposed (2 x/d) by inhalation to nominal concentrations of 20 and 100 mg PAA/m 3 throughout gestation. The atmospheres were generated from a 1% or 5% dilution of PAA (40%, H 2 O 2 14%, HOAc 27%). The authors reported a statisti cally significant retardation of foetal growth (body length and weight) at 100 mg PAA/m 3 , but no retardation at 20 mg/m 3 . No exposure related skeletal anomalies were observed. The health status of the dams wa s not reported (Kramer et al 1990). Exposure to a level of 100 mg PAA/m 3 would have been expected also to produce maternal toxicity. Because of uncertainty about the exposure levels a nd limited reporting, a reliable conclusion cannot be drawn from this study. No effects were reported from continuous breeding studies in rats, mice, gebrils and guinea pigs (Juhr et al , 1978).

Conclusion

Due to the absence of systemic ava ilability it is unlikely that the prod uct can reach the sex organs or the developing embryo or foetus under conditions of norm al handling. Therefore any further testing for reproductive endpoints is not warranted. 

### 2.6 Carcinogenicity

No valid carcinogenicity study is available. Due to the absence of systemic availability further testing for carcinogenicity is not warr anted. Furthermore, genotoxicity studies for PAA are available indicating that the substance was not genotoxic in most of the in vitro and the standard in vivo genotoxicity tests. It did not induce micronuclei in mouse micronucleus assays and was negative in two in vivo UDS assays in rats.

### MEPC 53/2/12 ANNEX Page 17

I:\MEPC\53\2-12.doc A tumour initiation-promotion study in mice has been published by Bock et al , 1975 and was discussed in ECETOC, 2001. In this study PAA was reported to act as a tumour promotor in mouse skin after initiation with 7,12-Dimethylbenz[a]anthracene (DMBA). It is likel y that this effect can be explained by the chronic irritation caused by the PAA treatment. The carcin ogenicity of hydrogen peroxide has been evaluated in EU, 2003. Further classical carcinogenicity studies are unlikely to reveal any additional insight to this endpoint.

Conclusion

Due to the toxicokinetics making it unlikely that Peraclean Æ Ocean is systemically available and the absence of a genotoxic potential in the majority of t he genotoxicity studies it is unlikely that the product has a relevant carcinogenic potential for humans and further testing is not warranted.



### 2.7 Mutagenicity

In vitro studies

The relevant in vitro studies are summarized in table 6.

Table 6: In vitro genetic toxicity tests

Test system Test organism, strain Composition (%) Dose (product) Metabolic activation Result Reference

PAA H 2 O 2 HOAcReversionassay (Ames test) Ta 98, TA 100, TA 102, TA 1535, TA 1537, TA 1538 4.5 25.5 6.7 7-4576 μg/plate Yes/No Neg Wallat, 1984a

Reversionassay (Ames test) TA 97, TA 98, TA 100, TA 1535 40 NS NS 0.3-200 μg/plate Yes/No Neg Zeiger et al , 1988

Chromosomal aberration Human lymphocytes 5.17 20 10 0.25-5 mg/ml Yes/No Neg Pos A Phillips, 1994b Chromosomal aberration Chinese Hamster V79 cells 10.7 NS NS 0.16-0.6 μl/ml (without S9) 0.5 to 1.18 μl/ml with S9 Yes/No Neg. CEFIC PAR, 2002a

Point mutation in mammaliancells, HPRTTest Chinese Hamster V79 cells 10.7 NS NS 0.16-0.6 μl/ml (without S9) 0.5-1.18 μl/ml (with S9) Yes/No Neg. CEFIC, PAR, 2002b

Unscheduled DNA synthesis Human lung fibroblasts WI-38 CCL75 31 4.7 NS 0.2-32 mg/ml No Neg Coppinger et al , 1983

DNA repair
assay Human lung fibroblasts WI-38 CCL75 42 5.5 NS 4-32 μg/ml No Neg Coppinger et al , 1983

A At highest, cytotoxic doses.

### MEPC 53/2/12 ANNEX Page 18

I:\MEPC\53\2-12.doc Equilibrium PAA (40%) proved to be negative in t he standard S. typhimurium reversion assay at concentrations of 0.3 to 200 μg PAA/plate. The hi ghest concentration was chosen after toxicity testing prior to the actual test and represents the dose t hat elicited toxicity or a dose immediately below (half-log dose intervals). In addition to rat liver S9 mi x, material from hamster tissues was also used as a metabolic system. With both protocols PAA was f ound to be devoid of mutagenic activity (Zeiger et al

, 1988).

Strain TA 102 was included in anothe r reversion-assay with PAA. In strain TA 102, PAA up to cytotoxic concentrations produced only a slight increase of reverted colonies (up to 30%) and the test substance was judged to be non-mutagenic. Total inhibition was achieved in concentrations exceeding 183 and 915 μg PAA/plate (depending on the stra in and activation system). With re gard to cytotoxicity no marked differences, i.e. reduced sensitivity, were observed in strain TA 102 (Wallat, 1984a). The concentrations that were reached in this assay were relatively high compared with the results pr esented by other authors. Studies in bacteria or yeasts with low reliability were conducted by Agnet et al , 1977 and Dorange et al

, 1974. In these studies equivocally positive effect s were reported in bacteria (S. typhimurium LT-2, TA 1978) and negative results in S. cervisia D7 cells.
Two independent well-documented expe riments on the potential of PAA to induce structur al chromosomal aberrations in human lymphocytes in the presence a nd absence of S9 mix (metabolic activation system) were conducted with equilibrium PAA (5.17%). Cells were treated at 0. 25, 0.5, 1.0, 2.0 and 2.0 mg PAA/ml for 20 hours in the fi rst experiment and with 0.25, 0.5, 0.75, 1.0 and 1.5 mg/ml in the second experiment. Treatment in the presence of S9 mix was carried out at 0.31, 0.63, 1.25, 2.5 and 5.0 mg/ml in both experiments and was limited to 3 hours. PAA revealed positive results only in the highest moderately cytotoxic doses, which reduced the mitotic index to 44.5 ñ 63 % without S9 mix and to 61 ñ 69 % with S9 mix (Phillips, 1994b). It is specul ated that the cytotoxicity and genotoxicity exerted by PAA is a result of the same mechanism at the cellular level, e.g. production of reactive oxygen species which are not detoxified at higher concentrations. In another chromosomal aberration test conducted according to recent OECD guidelines and GLP with 10.7% PAA. Two independent experim ents with 2 parallel cultures were performed in Chinese hamster V79 cells in the presence and absence of rat liver S9 mix. The dose selection was made in a pre-test for cytotoxicity. On the basis of this range finding study the concentrations in the main test were 0.09, 0.13, and 0.2 μl/ml in the absence and 0.13, 0.2 and 0.3 μl/ml in the presence of S9. The cells were treated for 4 hours in both experiments. Harvesting of the ce lls was performed at 18 hours in the first experiment and at 18 and 26 hours in the second experiment in t he absence of S9 and after 14 or 22 hours in the presence of S9. In the second experiment for the la ter harvest time only the highest concentration was evaluated. 100 metaphases per cu lture (200 per experimental point) were analysed for chromosomal aberrations. A reduced degree of confluence indicated cytotoxicity at a concentration of 0.13 μl/ml in the absence of S9 and at 0.2 μl/ml in the presence of S9. At 0.3 μl/ml the cells were completely damages without S9. No increase in chromosomal aberrati ons was observed at any of the test points in the presence and absence of S9. (CEFIC PAR, 2002a). An in vitro gene mutation test (HPRT test) was perfo rmed in Chinese Hamster V79 cells with 10.7% PAA in the presence and absence of rat liver S9 mi x. Two independent experiments were performed. Concentration selection was made following a range finding test for cytotoxicity. On the basis of the range finding study the concentrations in the main test were 0.16, 0.22, 0.31, 0.43, 0.5, and 0.6 l/ml in the absence of S9 and 0.5, 0.6, 0.7, 0.84, 0.98, and 1.18 μl/ml. The cells were exposed for 4 hours and maintained in exponential growth for 9 days. Cytoto xicity was evaluated by determining the plating efficiency on day 1 after treatment. At the end of t he expression period cells were cultured in selective medium for the determination of mutant frequency and in non-selective medium for the determination of plating efficiency. Cytotoxicty was observed from conc entrations of 0.22 μl/ml in the absence of metabolic activation and from 0.98 μl/ml with S9 (50% reduction in plating efficiency at 0.98 μl/ml). No statistically significant or dose related increase in mutation freq uency was observed both with and without S9 in both experiments. (CEFIC, PAR, 2002b).
The possible induction of Unscheduled DNA Synthesis by PAA was investigated in human diploid foetal lung cells (Coppinger et al , 1983). Concentrations of 0, 0.2, 0.5, 1, 2, 4, 9, 16, 24 and 32 μg PAA/ml were used. Positive controls were included. No statisti cally significant increase of UDS compared to solvent (water) controls was detect ed using autoradiography. In this assay 32 μg PAA/ml was clearly cytotoxic (50% survival). Using the same test system a nd controls, a DNA repair assay (three independent experiments) was conducted. The assay was negative at all dose levels. At the highest concentration normal DNA replication was considerably reduced (Coppinger et al , 1983).

### MEPC 53/2/12 ANNEX Page 19

I:\MEPC\53\2-12.doc In vivo studies

In a mouse micronucleus test with equilibrium PAA (5 .17%), groups of 15 male and 15 female CD-1 mice were given single oral doses (0, 8, 35 and 150 mg PAA/kg) by gavage. Positive control groups were included. There were no significant differences in the frequency of micronuclei in polychromatic or normochromatic erythrocytes between mice treat ed with PAA and the untreated controls. PAA did not induce a dose-related decrease in the proportion of po lychromatic erythrocytes, indicating a lack of toxicity to the bone marrow. No clinical signs were reported (Blowers, 1994a).

In a micronucleus test conducted with equilibrium PAA (4.5%), the te st solution was administered by gavage to groups of 7 male and 7 female CF21/W68 mi ce. The animals received two doses of 200, 400, and 800 mg PAA/kgbw/d at 0 and 24 hou rs. A positive contro l was included . Dose-dependent clinical signs of toxicity were observed in all groups. No mo rtalities were recorded within the time frame of the investigation and no increased incidences of micronuclei were found. General bone marrow toxicity was detected as the inhibition of proliferation in the eryt hropoiesis since the ratio of normochromatic versus polychromatic erythrocytes was increased in the highest dose group (Wallat, 1984b).

An in vivo/ex vivo UDS assay was conducted in groups of 6 male F344 rats receiving doses of equilibrium PAA (5.17%) at 330 or 1,000 mg PAA/kgbw by gavage. Po sitive controls were in cluded. No significant increases in UDS (measured as net grain increase) were observed in both treated groups at either time. It was concluded that the tested PAA formulation was not genotoxic u nder the conditions of the study (Blowers, 1994b).

Another in vivo/ex vivo UDS assay in groups of 3 male Fischer rats receiving doses of 500, 1000 and 2000 mg/kg bw 5% equilibrium PAA (with 14 % hy drogen peroxide an d 17.6% acetic acid) positive controls were included. Expression times were 2-4 an d 12-16 hours. At least 150 cells were counted per animal. The dose selection was based on a pre-experiment to determine the maximum tolerated dose level. Nuclear and cytoplasmatic grain counts were obt ained from 50 cells per slide. The slides of the lowest dose were not evaluated as the 2 high doses could be satisfactory analysed. No increase of nuclear grain counts was observed at any of the doses an d time points tested. No significant increase in the percentage of cells in repair was detected at any dose and time point of PAA when compared with the respective controls. Moreover the frequency of cell s in the S-phase was low in the treated animals. The test substance did not reveal any genotoxic ac tivity under the test conditions employed. (CEFIC PAR, 2002c).

Conclusion

The weight of evidence in particula r in the more recent studies in in vitro experiments in mammalian cell cultures indicates that equilibrium peracetic acid does not have a significant genotoxic potential and did not cause gene mutations and chromosomal aberrations both in the presence and absence of a metabolic activation system. Only in one chromosomal aberration assay in vitro a positive result was obtained at highly cytotoxic concentrations. However, a recent in vitro chromosomal aberration assay did not show such an effect even at cytotoxic concentrations suggesting that the earlier findings may have been an artefact. PAA did not induce micronuclei in in vivo mouse micronucleus tests when administered by gavage up to the maximum tolerated dose levels and did not induce unscheduled DNA synthesis in two recent rat in vivo/ex vivo UDS assays when administered by gavage up to the maximum tolerated dose level. Therefore it can be concluded that Peraclean Æ Ocean is unlikely to have a genotoxic or mutagenic potential.

3 Data on environmental fate and effect under aerobic and anaerobic conditions (4.1.2.3; 5.3.4; 5.3.5)



### 3.1 Modes of degradation

Abiotic degradation of e quilibrium peracetic acid An abiotic degradation study according to EEC92/69 report ed half lives of peracetic acid at 25 °C of 48 h at pH 7 and pH 4. At pH 9 the hal f life was lower than 3.6 hours (Pierre et al , 1995). The decomposition of Degaclean 150 containing ab out 21% of hydrogen peroxide and 15% of peracetic acid was studied in synthetic seawater (Degussa, 2 000). At a salinity of 2% (w/w) 105 mg/L were degraded with a half life of approximately 20 minutes for peracetic acid. At a salinity of 3.3 % (w/w) the half life of PAA was about 7 minutes. Degradation of hydrogen peroxide u nder these c onditions was

### MEPC 53/2/12 ANNEX Page 20

I:\MEPC\53\2-12.doc slower. About 20% were degraded within 35 min or 25 min at salinity of 2% or 3.3% respectively. It should however, be consi dered that natural sea wate r will contain transition metals, organic compounds and organisms that will most pro bably lead to a more rapid degrada tion of hydrogen peroxide and PAA.

Abiotic degradation of hydrogen peroxide

H 2 O 2 is subject to various reduction or oxidation processes in the environment and decomposes into water and oxygen at rates which depend on contact with catalytic materials and other factors. Common catalysts are dissolved transition metals (e.g. cop per, iron, manganese), solid metals (e.g. platinum, osmium or silver), solid metal oxides and hydroxid es (e.g. manganese, iron, copper oxides), activated carbon and enzymes. H 2 O 2 is also decomposed by alkaline impurities sufficient to raise the pH to 7 or above. Heat and sunlight can induce photochemical decomposition with the formation of free radicals (ECETOC, 1997)
Biotic degradation of equilibrium PAA

Peracetic acid was readily biodegradable in a study according to OECD 301 E. More than 70% degradation was obtained after 28 days and the 10 day window was met (Richterich and Gode, 1986).
During an activated sludge respirat ion inhibition test at a concentration of 0.3 to 100 mg PAA/l the degradation during the test was determined. The hal f life was < 3 minutes at concentrations between 0.3 and 30 mg/ml and about 30 minutes at 100 mg/L. (De Groot, 2003).
Biotic degradation of hydrogen peroxide

Hydrogen peroxide was evaluated on the basis of a number of different data to be readily biodegradable fulfilling the 10 day window in sewage treatment plants and under environmental conditions. Typical half lives in sewage treatment plants were 2 minutes, half liv es in the environment varied from a few hours to typically 1 to 3 days up to 5 days. Higher hydrog en peroxide concentrations can slower the rate due to the inhibitory effect of hydrogen pe roxide on microorganisms (EU 2003).



### 3.2 Bioaccumulation, partition coefficient, octanol/water coefficient Partition coefficients

For Peracetic acid a log Kow of ñ 0.52 Byers, 1998) and for hydrogen peroxide a log Kow of -1.5 (EU, 2003) was reported. The low octanol water partition coefficient indicates a low potential for bioaccumulation.
Partitioning between environmental compartments

The theoretical distribution of PAA has been estimated using the fugacity model of Mackay, Level 1. (ECETOC, 2001). T he calculations were conducted for the equi librium grades of PAA, using the partial vapour pressure of PAA in a comp osition comparable to Peraclean Æ Ocean. According to the model, the majority of PAA (99.3 to 99.9%) released into t he environment enters the water phase, while the remainder is found in air. No partitioning into soil, suspended matter or biota is expected (Table 4). A risk characterisation for Ballast water treatment can therefore be restricted to the aquatic compartment.

Table 4: Distribution of PAA between Environmental Compartments at 25°C (ECETOC 2001)

Compartment Distribution(%)

Air 0.29

Water 99.71

Soil 0.00

Sediment 0.00 Suspended matter (aquatic) 0.00

Biota 0.00

### MEPC 53/2/12 ANNEX Page 21

I:\MEPC\53\2-12.doc The same applies for hydrogen peroxide. An equilibr ium partitioning has been calculated according to Mackay level I at 20°C, a vapour pressure of 3 hPa and a Henryís law constant of 7 10 -4 Pa m 3 /mol. (see table 5)(EU 2003).

Table 5: Distribution of hydrogen peroxide between environmental compartments at 20°C (EU, 2003)

Compartment Distribution (%)

Air 0.3 x 10 -3

Water 99.98

Soil 0.01

Sediment 0.01

3.3 Persistence and identification of the main metabolit es in relevant media (ballast water, marine and fresh water)

The primary metabolites of Peraclean Æ Ocean in all relevant media are acetic acid, oxygen and water. Hydrogen peroxide stability is greater than that of Peracetic acid, but in the environment it will ultimately be degraded by biotic and abiotic processes to oxy gen and water. Acetic acid is a product of the intermediary metabolism and will ultimately be oxidised to CO 2 and water or integrated into biomolecules by organisms.

3.4 Reaction with organic matter

PERACLEAN Æ Ocean is a chlorine-free oxidizer. In consequence the natural traces of halogenated organic compounds in the ballast water are not increased. Side reactions generating epoxides and organic peroxides are not likely due to the short half-life of PAA and the conditions of ballast water treatment (low PAA concentration, low temperature, non- presence of catalyti c compounds, neutral pH-level).

Measurements by gas chromatography /mass spectrometry using river wa ter showed that the disinfection by-products were carboxylic acids, which ar e not recognized as mutagenic (Monarca, 2002)

3.5 Potential physical effects on wildlife and benthnic habitats

Not applicable. As Peraclean Æ Ocean is not persistent in the envir onment it is not expected to have any effects on wildlife or benthnic habitats.

3.6 Potential residues in seafood

As Peraclean Æ Ocean is not systemically available to or ganisms and is readily metabolised to oxygen, water and acetic acid no potential residues in seafood are to be expected.

3.7 Known interactive effects



### None.

### MEPC 53/2/12 ANNEX Page 22

I:\MEPC\53\2-12.doc 4 Physical and chemical properties (4.1.2.4)

The relevant physical chemical properties are summarised in table 6.

Table 6: Physical chemical properties

Property Value Reference/comment
Physical state Clear, colourless liquid Melting point Ca. -50°C ECETOC (2001) Boiling point > 60°C ECETOC (2001) Relative density 1,150 kg/m 3 at 20°C ECETOC (2001) Vapour pressure 110 hPa at 20°C ECETOC (2001)/ Partial vapour pressure, peracetic acid (PAA), Water solubility 1,000 g/kg Swern, 1970

Partition coefficient noctanol/water (log value) -0.52 (measured value) Byers, 1998/ for PAA

Partition coefficient noctanol/water (log value) -1.5 EU, 2003 / for hydrogen peroxide

pKa 8.2 at 20°C Swern, 1970
Flash point Ca. 96°C Autoignition temperature Ca. 265°C
Saturated Vapour concentration Ca. 14% ECETOC, 2001

Corrosion

In a study conducted with fresh water (as a worst case condition) plus Peraclean Æ Ocean neither a significantly higher corrosion nor corrosion nor a dam aging of the usual Epoxy-based ballast water tank coating was observed. The trials were performed using the normal dosage of 150 ppm PERACLEAN Æ Ocean and the double dosage of 300 ppm, respectively. In contrast to the application onboard of a ship, where PERACLEAN Æ Ocean degrades rapidly Peraclean Æ Ocean was refreshed/replaced regularly to maintain a constantly high concentrati on during the testing period of 30 days.

Even under these extreme conditions (according ìTesting Method for Evaluation of Painted or Coated Specimens Subjected to Corrosive Environmentsì, ASTM D1654-92) PERACLEAN Æ Ocean did not significantly increase corrosion (Degussa 2004a). No influence on the corrosion of steel St 37-2 was observed in a corrosion test with 3.3% sea-salt and 500 ppm Peraclean Æ Ocean stored for 14 days at temperatures between 25 and 27°C (Degussa, 2003). Various corrosion tests with different grades of peracetic acid (5, 15, 35%) and different stainless st eels under various test conditions showed that all grades were corrosion resistant (Degussa, 2001b).

5 Analytical methods at environmentally relevant concentrations (4.1.2.5; 5.3.15)

6 Toxicity testing of treated ballast water (5.2)

To simulate conditions of ballast water treatment, a 150 mg/L solution of Peraclean Æ Ocean was prepared in a 20 L volume of seawater with no headspace in a plastic container. This solution was placed in the dark at 8°C for 48 hours. After the incubation time the solution was eval uated for its toxicity to bivalve larvae ( Mytilus galloprovincialis) , < 4 h post fertilisation. The te st duration was 48 hours (Degussa, 2005b).

### MEPC 53/2/12 ANNEX Page 23

I:\MEPC\53\2-12.doc This test system was chosen as the only reference te st system, as it proved to be the most sensitive system in a pre-test with Peraclean Æ Ocean using different marine organisms and different study durations (Degussa, 2005a). The resu lts of this test were compared wi th those of a freshly prepared sample of 150 mg/L Peraclean Æ Ocean. As Peraclean Æ Ocean also reduces the pH as a result of the presence and formation of acetic acid in the solution one pH adjusted test sample was also included to determine the extend of residual toxicity that could be caused by a decrease in pH. In order to determine whether residual toxicity could be due to hydrogen pe roxide, a bivalve larval development test was also conducted with hydrogen peroxide at concentrations of 0.078 to 5 mg/L . Additionally the degradation of residual toxicity after the addition of catalase was tested, by adding catalase to a sample of 150 mg/L Peraclean Æ Ocean stored in the dark for 48 h at 10°C. Hydrogen peroxide concentrations were measured in all samples (detection limit: < 0.1 mg/L).

Results:

Degradation of toxicity with time

The EC 50 of the untreated sample was 2.4 mg/L, the NOEC was 1.56 mg/L. After 48 h of storage the EC 50 was 18.6 mg/L without pH adjustment and 17.6 mg/L with pH adjustment, the corresponding NOEC were 12.5 and 6.25 mg/L respectively. The results indicate an approximately 86% reduction of the toxicity during the 48 h storage period. Adjustment of pH did not decrease toxicity and the ac idity was not responsible for the residual toxicity. Toxicity of hydrogen peroxide

The EC 50 for normal surviving larvae was 1.58 mg/L, t he NOEC was 1.25 mg/L. The measured hydrogen peroxide concentration 48-hour de graded sample of Peraclean Æ Ocean was 11 mg/L. The results indicate that there was sufficient hydrogen peroxide present in the aged solution to explain the residual toxicity. The residual toxicity was diminished by the addition of catalase to the 48-h degraded sample of Peraclean Æ Ocean, thus providing further evidence that the residual toxicity was caused by hydrogen peroxide.
7 Exposure assessment



### 7.1 Environmental exposure assessment (5.2.7)

Entries of Peraclean Æ Ocean into the aquatic environment can only occur when Ballast water is exchanged. Preliminary experiments on the residual concentrations of peracetic acid and hydrogen peroxide in treated ballast water samples have been performed during field trials (Degussa, 2004c). Ballast water treatment was performed with the recommended equipment and concentrations of Peraclean Æ Ocean. All treated ballast water was direct ly collected in 6 simula ted ballast water tanks (20 t tank containers). Samples were taken at di fferent reaction times and the content of the active ingredient was analysed. A complete decay of the ac tive ingredient peroxyacetic acid was observed after 10 to 15 hours. Hydrogen peroxide was degraded to about 20 mg/l after 5 to 10 hours and to 15 mg/l after up to 50 hours. The toxicity testing of treat ed ballast water confirmed that a concentration of about 10 mg/L of hydrogen peroxide was still present after 48 h of storage of a solution of 150 mg/L Peraclean Æ
Ocean in seawater at 8 to 10°C. When released into the environment resid ual hydrogen peroxide is further diluted and also expected to be rapidly degrade d by the various routes of degradation. Preliminary calculations using the MAMPEC model developed by the Dutch Institute for Environmental Studies indicate that the concentrations of hydrogen pero xide in commercial harbours will remain below the conservative PNEC for hydrogen peroxide of 10 μg/L (maximum concentration was 4.9 μ/l), while the PAA concentration was predicted to be ab out 6 orders of magnitude lower (4.1 10E-6 μg/L ) and can be regarded as negligible.



### 7.2 Human exposure assessment

Due to the automated process in ballast water applic ation and dosing human exposure is unlikely to occur during the treatment operations. Very short dermal ex posure would be possible when the containers are connected to the dosing system. In any case for su ch operations and for accidental situations personal protective equipment, protective gloves, goggles and if needed respiratory protection and full protective

### MEPC 53/2/12 ANNEX Page 24

I:\MEPC\53\2-12.doc suits are recommended and should be provided. Safety training is also offered by the producer of Peraclean Æ Ocean.

8 Risk characterisation (5, 5.3)



### 8.1 Screening for persistency, bio-accumulation and toxicity (5.1)

The active components of Peraclean Æ Ocean do not fulfil the PBT criteria as outlined in EC, 2003a:

Criterion PBT criteria P Half-life > 60 d in marine water or > 40 d in freshwater * or half-life > 180 d in marine sediment or > 120 d in freshwater sediment * or readily biodegradable B BCF > 2,000 or log Kow > 4.5 T Chronic NOEC < 0.01 mg/l or CMR or endocrine disrupting effects

Persistence (P)

Due to the ready biodegradability and the abiotic degradation of the active components of Peraclean Æ Ocean, it can be concluded that it is not persistent in the environment. Environmental half lives are in the order of hours or a few days at maximum.

Bioaccumulation (B) Due to the high water solubility and the low octanol wate r partition coefficients of the active components of Peraclean Æ Ocean, there is no concern for a possible bio-accumulation potential of the product (log Kow below 0).
Toxicity (T)

The active components of Peraclean Æ Ocean do not have NOEC levels below 0.01 mg/l and there are no indications for relevant effects concerning carc inogenicity, mutagenicity or reproductive toxicity.



### 8.2 Assessment of possible reaction with organic matter (5.3.3)

8.3 Rate of abiotic and biotic degradati on and degradation products (5.3.4, 5.3.5)

The active ingredients of Peraclean Æ Ocean, PAA and hydr ogen peroxide degrade rapidly in the environment. Abiotic degradation half lives in seawater are in the range of several minutes for Peracetic acid, and up to 48 h in freshwater at pH 4 to 7. For hydrogen peroxide half lives of several hours up to 5 days were predicted under environmental conditions. Th e half life will be more rapid in the presence of transitions metals, metal oxides and hydroxides, at pH Values > 7. Heat and sunlight can induce photochemical decomposition. However it should also be considered that hydrogen peroxide is also generated photochemically in water systems. In se awater the limited amount of available natural concentrations ranged from 0.5 to 14 μg hydrogen peroxide/l, in lakes values > 100 μg/l were reported.
Both PAA and hydrogen peroxide are r eadily biodegradable with half liv es with sewage treatment plant inoculum of 2 to 3 minutes if the toxicity levels are not superseded. Degradation of Peraclean Æ Ocean results only in the formation of oxygen and water as well as acetic acid that can be readily metabolised yielding carbon dioxide and water. No persistent metabolites are formed.

### MEPC 53/2/12 ANNEX Page 25

I:\MEPC\53\2-12.doc 8.4 Partition coefficients (5.3.6)

From the Octanol-Water partition coefficients of the active substa nces PAA and hydrog en peroxide, the organic carbon normalised distribution coefficient can be derived using the following equations according to the EU technical guidance docum ent on risk assessment (EC, 2003). For organic acids the following equation can be used:
log Koc = 0.6 log Kow + 0.32 For Peracetic acid using a log Kow of -0.52 a log Koc of 0.008 (Koc = 1.02 L/kg) is derived. For hydrogen peroxide the following equation for non-hydrophobics may be used:
log Koc = 0.52 log Kow + 1.02 Using that equation a log Koc of 0.2 (Koc = 1.58 L/kg) was derived for hydrogen peroxide (EU, 2003).
Both Koc values indicate a negligible absorption to organic matter which would also indicate a very low adsorption potential to soil and sediment. This is c onfirmed by the Mackay level I equilibrium distribution (see chapter 3.2).

8.5 Bioaccumulation potential and secondar y poisoning potential (5.3.7, 5.3.10)

Due to the low Octanol-water partit ion coefficients of the active ingr edients (log Kow of PAA = -0.52, of hydrogen peroxide -1.5) Peraclean Æ Ocean has no bioaccumulation potential and thus an assessment of secondary poisoning is redundant.

8.6 Assessment of sediment species (5.3.11)

As the Koc values of the active ingredients PAA and hydrogen peroxide are ve ry low (1.02 L/kg and 1.58 L/kg) an assessment of s ediment poisoning is redundant.

8.7 Comparison of the results of the effect assess ment with the results of the discharge toxicity testing. (5.3.14)

Discharge toxicity testing was performed with the mo st sensitive of the marine species determined in a pre-study, Mytilus galloprovincialis larvae < 4 h post fertilisation. Th e test duration was 48 hours. (Degussa, 2005b). After 48 h the toxicity was reduced to 80% of the original toxicity. The residual toxicity could be completely attributed to a residual content of hydrogen peroxide. This is in accordance with the findings that PAA from Peraclean Æ Ocean at the concentration of 150 mg/L recommended for Ballast water treatment is completely degraded already afte r 10 to 15 hours during Ballast water treatment, while hydrogen peroxide degradation is slower and it is still present in the treated solution at a concentration of 10 to 15 mg/L after 48 to 50 hours. When released into the environment resi dual hydrogen peroxide is further diluted and also expected to be rapidly degrade d by the various routes of degradation. Preliminary calculations using the MAMPEC model of the Dutch Inst itute for Environmental Studies (IVM) indicate that the concentrations of hydrogen peroxide in commerci al harbours will remain below the conservative PNEC for hydrogen peroxide of 10 μg/L (max imum concentration was 4.9 μ/l), while the PAA concentration was predicted to be about 6 orders of magnitude lower (4.1 10E-6 μg/L) and can be regarded as negligible. Taking into account that the predicted hydrogen peroxide concentrations are in the range of the naturally occurring concentrations of hydrogen perox ide in the environment it can be concluded that those discharges do not constitute a risk to the environment.

8.8 Human health risk assessment

Due to the technology used humans will not be exposed to Peraclean Æ Ocean during ballast water treatment on the ships. During the recommended oper ating procedures the possibility of exposure will be minimal and limited to short term contact when connec ting the storage tank to the dosage line. Direct contact during this operation is minimized by the recommended handling procedures that include the appropriate personal protective equipment, gloves, g oggles and if needed respiratory protection. As the main hazard of the product is due to local irritation/corrosion at the site of contact and there is no immediate concern for mutagenicity or carcinogeni city, the product can be handled safely in this application.

### MEPC 53/2/12 ANNEX Page 26

I:\MEPC\53\2-12.doc Appendix

Results of Field Trials Carried ou t in Kiel Harbour in November 2002

Abbau des Desinfektionsmittels in den La g ertanks Messung am 26.11.2002

77777 6 ,9 6 ,9 6,9 6 ,7 6,7 6 ,8

3 0

22,4 2 4

2 1,2

16 ,8

15 14 ,1 13 ,8

12 ,3 12 ,9

2,5 2 2 ,5

17,6 16,8

13 ,6

9 ,9 9 ,6 9,6

5,1 4,2 3 ,3

0,1 0 5 10 15 20 25 30 35

Zeit nach der Behandlung pH-Wert [-] H2O2-Gehalt [mg/l]
Peressigs‰ure [mg/l]
genehmigter minimaler pH-Wert [-]
genehmigter maximaler H2O2-Gehalt [mg/l]

Abbau des Desinfektionsmittels in den La g ertanks Messung am 28.11.2002

6 ,7 6 ,8 7 7,2 7,2 6 ,9

17,2

15,1

13 ,8

9 ,6

8 ,1

1,4 9 ,9

6,7

5,9

4 ,8 4 ,3

0 0 5 10 15 20

Zeit nach der Behandlung pH-Wert [-] H2O2-Gehalt [mg/l]
Peressigs‰ure [mg/l]
genehmigter minimaler pH-Wert [-]
genehmigter maximaler H2O2-Gehalt [mg/l]



### ___________