Translation Statement 

Institute ofEcotoxicology Co., Ltd. 

Sponsor: 

Title: Bioconcentration Test of in Carp, Cyprinus carpio 

Test No.: C-186 

I, the undersigned, hereby declare that this report provides a correct English 

translation of the Final Report (Study No. C-186, issued on March 1, 2011). 

Date May 15,2012 

Study Director k.ka~l Kenji Kaneko 
Final Report 

Bioconcentration Test of in Carp, Cyprinus carpio 

Report No. E4 -11006·C186·NS 

March 1, 2011 

Institute ofEcotoxicology Co., Ltd. 
Table of Contents 

GLP Statement 

Quality Assurance Certificate 

Preface 

1. 

2. 

3. 

4. 

Title 

Sponsor 

Test facility 

Objective of the test 

5. Test methods 

6. 

7. 

8. 

9. 

10. 

11. 

12. 

13. 

14. 

15. 

GLP applied 

Name of the test substance 

Test periods 

Test results 

Evaluation, discussion of the results and conclusion 

Storage ofthe test source materials 

Environmental factors assumed to have an effect on reliability of the data 

Personnel related to the test and their assigned positions 

Name and address of scientists having contributed reports to the final report 

Approval of the final report 

Main Subject 

1. 

1.1 

1.2 

1.3 

1.4 

1.5 

2. 

2.1 

2.2 

2.3 

Test substance 

Name 

Structural formula 

Supplier and Lot No. 

Identification 

Physico-chemical properties 

Test methods and analytical methods 

Acute toxicity test (96h LC50) in Zebrafish (Danio rerio) for selecting 

the substance concentration 

Performing a bioconcentration test 

Analyses of fish and test water 

Page 

1 

11 

1 

1 

1 

1 

1 

2 

2 

2 

2 

3 

3 

4 

4 

5 

5 

6 

6 

6 

6 

6 

6 

7 

7 

7 

8 

12 
2.3.1 Analytical method for fish 12 

2.3.2 Analytical method for test water 14 

2.3.3 Measurement methods 16 

2.4 Calibration curve by liquid chromatography mass spectrometry 16 

2.4.1 Preparation of standard solutions 16 

2.4.2 Preparation of calibration curve 17 

2.5 Recovery test and blank test for 17 

2.5.1 Recovery test from fish for 17 

2.5.2 Recovery test from test water for 18 

2.6 Quantifiable limit 19 

2.6.1 Quantifiable limit for fish 19 

2.6.2 Quantifiable limit for test water 20 

2.7 Preliminary test 20 

2.8 Schedule of analyses of fish and test water 21 

3. Test results 22 

3.1 Concentrations of in test water 22 

3.2 Bioconcentration factors (BCFs) 23 

3.3 Evaluation, discussion of the results and conclusion 24 

Table of contents for references 25 

This report includes 90 pages 
GLP Statement 

Institute ofEcotoxicology Co., Ltd. 

Sponsor: 

Title of the test: Bioconcentration Test of in Carp, Cyprinus carpio 

Test No.: C-186 

The aforementioned test was conducted in compliance with following GLP principles: 

"Standards Concerning Test Facilities Performing Tests Concerning New Chemical 

Substances" on Japanese GLP (Yakushokuhatsu No. 1121003, Heisei 15 • 11 • 17 Seikyoku 

No.3 and Kanpokihatsu No. 031121004, Joint notification ofDirector General of 

Pharmaceutical and Food Safety Bureau in Ministry of Health, Labor and Welfare, Director 

General of Manufacturing Industries Bureau in Ministry ofEconomy, Trade and Industry and 

Director General of Environmental Policy Bureau in Ministry of the Environment) (dated 

November 21, 2003) 

"OECD Principles of Good Laboratory Practice" (as revised in 1997). 

March 1, 2011 

Study Director: Signed in original 

Kenji Kaneko 
Report No. E4 -11006· C186 · NS TestNo. C-186 

Quality Assurance Certificate 

Title: Bioconcentration Test of in Carp, Cyprinus carpio 

Sponsor: 

Study Director: Kenji Kaneko 

Study Personnel: Kenji Kaneko 

Test Facility: Institute ofEcotoxicology Co., Ltd. 

We certify that the present final report describes the test methods accurately and the 

contents were performed according to the test protocol and standard operation procedures, 

and the original data are accurately presented. The inspection and audit were performed 

as follows. 

Dates performed 

Status of performance Water/fish analysis on Day 7 from the start February 2, 2011 

of test inspected of exposure 

Water/fish analysis on Day 14 from the February 9, 2011 

start of exposure 

Water/fish analysis on Day 21 from the February 16, 2011 

start of exposure 

Test protocol audited Protocol audited January 12, 2011 

Protocol (revision) audited January 18,2011 

Final report audited Report audited March 1, 2011 

Institute of Ecotoxicology Co., Ltd., Department of Quality Assurance Unit 

Quality Assurance Director: Signed in original 

Mikiyasu Goto 

Signature date: March 1, 2011 

(Inspections of study conduct, protocol and final report) 

Quality Assurance : -~@ed in orig"--in_a_l ______ _ 

Quality Assurance : 

Mitsuko Takamatsu 

Signature date: March 1, 2011 

(Inspections of protocol and final report) 

Signed in original 

Miyuki Kurihara 

Signature date: March 1, 2011 

(Inspection of final report) 

11 

Dates reported to Test 

Facility Management 

and Study Director 

February 2, 2011 

February 9, 2011 

February 16,2011 

January 12, 2011 

January 18, 2011 

March 1, 2011 
1. Title 

2. Sponsor 

Final Report 

-PrefaceBioconcentration Test of in Carp, Cyprinus carpio 

3. Test facility Institute ofEcotoxicology Co., Ltd. 

2-28-1 Yoshino-cho, Kita-ku, Saitarna City, 331-0811 Japan 

Representative 

Test Facility Management 

4. Objective of the test 

Mikiyasu Goto 

Tadayoshi Kan 

The objective ofthis test is to determine the degree ofbioconcentration of the test 

substance in fish in order to file for a Chemical Substance Control Law in Japan (No. 

117 ofOctober 16, 1973). (abbreviated to 'CSCL' below) 

5. Test methods 

The methods used in this test were based on "Test Methods Concerning New Chemical 

Substances< Bioconcentration Test of Chemical Substances in the Bodies ofFish and 

Shellfish>" (Yakushokuhatsu No.l121002, Heisei 15 • 11 • 13 Seikyoku No.2 and 

Kanpokihatsu No. 031121002, Joint notification ofDirector General ofPharrnaceutical 

and Food Safety Bureau at the Ministry of Health, Labor and Welfare, Director General 

ofManufacturing Industries Bureau in Ministry of Economy, Trade and Industry and 

Director General of Environmental Policy Bureau in Ministry of the Environment, 

Japan) (dated November 21, 2003) and OECD Guidelines for the Testing of Chemicals, 

"No. 305 Bioconcentration: Flow-through Fish Test" (adopted on June 14, 1996). 
6. GLP applied 

7. 

8. 

Japanese Chemical Substance GLP: "Standards Concerning Test Facilities Performing 

the Tests Concerning New Chemical Substances" (Yakushokuhatsu No.l121003, 

Heisei 15 • 11 • 17 Seikyoku No.3 and Kanpokihatsu No. 031121004, Joint 

notification of Director General of Pharmaceutical and Food Safety Bureau at the 

Ministry of Health, Labor and Welfare, Director General of Manufacturing Industries 

Bureau in Ministry ofEconomy, Trade and Industry and Director General of 

Environmental Policy Bureau in Ministry ofthe Environment)( dated November 21, 

2003) 

OECD GLP: "the OECD Principles of Good Laboratory Practice" (as revised in 1997). 

Name of the test substance 

Chemical name 

Abbreviation 

CAS No. 

Test periods 

(1) Study initiation date January 12, 2011 

(2) Duration of the test 

Date of arrival of fish August 23,2010 

Acclimation completion date January 26,2011 

Experiment starting date January 26,2011 

Experiment completion date February 23, 2011 

(3) Study completion date March 1, 2011 

9. Test results 

The bioconcentration test of the test substance in carp, Cyprinus carpio was 

conducted. The test substance is a salt consisting of 

, and 

dissociates completely in water. In this test, the bioconcentration of only was 

confirmed. The bioconcentration factors of during the 28-day exposure period are 

summarized in the next page as a table. 

2 
High exposure level Low exposure level 

Bioconcentration factor of (Test substance: 0.2 mgL-1

) (Test substance: 0.02 mgL- 1

) 

( : 0.193 mgL-1

) ( : 0.0193 mgL- 1

) 

Bioconcentration factors under a detectable limit under a detectable limit 

(BCF) (less than 1-fold) (less than 11-fold) 

10. Evaluation, discussion of the results and conclusion 

The test substance is a salt consisting of 

only was confirmed. Since 

In this test, the bioconcentration of 

listed as an existing chemical 

substance, is not high bioconcentration, the bioconcentration of is considered to be 

confirmed by analogy. 

The 96h LCso value of the test substance in Zebrafish (Dania rerio) was greater 

than 101 mgL-1

. Since the solubility in water was> 1.0 x 105 mgL-1

, two concentrations 

of the test substance in water were set up to be 0.2 mgL-1 ( 0.193 mgL-1

) for the high 

exposure level (Concentration Levell) and 0.02 mgL-1 ~ 0.0193 mgL-1

) for the low 

exposure level (Concentration Level2). According to the results of the bioconcentration 

test, the bioconcentration factors (BCFs) in both Concentration Levels 1 and 2 were less 

than 50-fold. Thus, a steady state was assumed to be achieved within 28 days of exposure. 

It was confirmed that the bioconcentration was under a detectable limit and the 

bioconcentration factors were less than 1-fold in Concentration Level 1 and less than 11-

fold in Concentration Level 2. Therefore, bioconcentration of 

anion of the test substance was confirmed to be low. 

11. Storage of the test source materials 

The following records and samples concerning the tests will be stored for 10 years after 

receiving notification based on the provisions of Chemical Substances Control Law 

(CSCL) (No. 117, 1973): Article 4, Paragraphs 1 or 2, Article 4-2, Paragraphs 2, 3 or 8, 

Article 5-4, Paragraph 2, Article 24, Paragraph 2 or Article 25-3, Paragraph 2, and the 

test substance, reference substance and other items will be stored for 1 0 years after 

receiving notification based on the provisions of the CSCL: Article 4, Paragraphs 1 or 

2, Article 4-2, Paragraphs 2, 3 or 8, Article 5-4, Paragraph 2, Article 24, Paragraph 2 or 

3 
Article 25-3, Paragraph 2 or the period wherein they can be stored stably without 

causing any interference with quality, which is at least one ofthe periods that is shorter, 

at the Institute of Ecotoxicology Co., Ltd.. Regarding the storage after these periods, it 

will be determined separately after discussion. 

(1) Protocols, records of modifications and changes 

(2) Records concerning the test substance, etc. 

(3) Experimental notes, raw data, charts 

(4) Test substance, reference substance, other items (stored at about 20°C in a dark place) 

(5) Inspection and audit records recorded by the personnel in charge of quality 

assurance regarding the present test 

(6) Final report 

(7) Other items that are required 

12. Environmental factors assumed to have an effect on reliability of the data 

No particular abnormalities were found. 

13. Personnel related to the test and their assigned positions 

Study Director: Kenji Kaneko 

Study Personnel: Kenji Kaneko 

• 96h LCso test 

• Establishment of analytical methods for test water and fish 

• Preparation of calibration curve for analysis of test water and fish 

• Recovery tests of test water and fish 

• Preliminary test to arrange the study period and the sampling schedule 

• Analyses of test water and fish during the experimental period 

• Preparation and maintenance of test solution during the experimental period 

• Measurement of lipid contents in fish body 

• Measurement of IR 

• Observation of the conditions of fish 

• Measurement of water environment 

Final report prepared by Kenji Kaneko 

4 
14. Name and address of scientists having contributed reports to the final report 

Not applicable 

15. Approval of the final report 

Study Director: Signed in original 

Kenji Kaneko 

Date signed: March 1, 2011 

5 
Main Subject 

1. Test substance 

In the present report, the test substance has the following name and structural formula 

(based on the material provided by the sponsor). 

1.1 Name 

Chemical name: 

Abbreviation: 

CAS No.: 

1.2 Structural formula 

The abbreviation words were used in this test as shown below. 

Molecular formula: 

1.3 Supplier and Lot No. 

(1) Supplier 

(2) LotNo. 

1.4 Identification 

It was identified by the agreement of theIR spectrum provided by the sponsor with the 

IR spectrum measured at our institution at the time ofthe start of the test. Further, the 

stability of the test substance during the test period was confirmed by the agreement of 

theIR spectrum at the end of the test with theIR spectrum measured at the start of the 

test (see, Ref. 12). 

6 
1.5 Physico-chemical properties (based on the data provided by the sponsor) 

Appearance*: 

Purity*: 

Solubility (25°C) measured at Institute of Ecotoxicology: 

water > 1.0 x 105 mgL-1 

dimethyl sulfoxide > 1.0 x 105 mgL-1 

acetone > 1.0 x 105 mgL-1 

acetonitrile > 1.0 x 105 mgL-1 

methanol > 1.0 x 105 mgL-1 

dichloromethane 5.1 x 101 mgL-1 

n-hexane 6.0 X 1 o-2 mgL-1 

2. Test methods and analytical methods 

The methods used in this test were based on the Japanese Guideline, "Test Methods 

Concerning New Chemical Substances < Bioconcentration Test of Chemical 

Substances in the Bodies ofFish and Shellfish>" (Yakushokuhatsu No. 1121002, 

Heisei 15 • 11 • 13 Seikyoku No. 2 and Kanpokihatsu No. 031121 002) (dated 

November 21, 2003) 

and 

the OECD Guidelines for Testing of Chemicals, "No. 305 Bioconcentration: Flowthrough Fish Test" (adopted on June 14, 1996) 

2.1 Acute toxicity test (96h LC50) in Zebrafish (Danio rerio) for selecting the 

substance concentration (SOP 2.6) 

Based on the results of a preliminary test, the concentrations of the test substance in 

test water of a 96h LC50 test were selected. The 96h LC5o value was determined from 

the test result (the test was performed with reference to the method described in 

OECD Test Guidelines 203). 

(1) Test fish Zebrafish (Danio rerio) Lot No. 101202 

Supplier Kamihata Fish Ind., Ltd. 

Date received 

Mean total length 

Mean bodyweight 

292-1 Chigusa-cho, Hanamigawa-ku, 

Chiba City, 262-0012 Japan 

December 2, 2010 

3.2 ± 0.1 em 

0.24 ± 0.03 g 

Acclimation period December 2, 2010 - December 20, 201 0 

Receiving and acclimatization procedures were based on SOP 2.1 and SOP 2.5. 

7 
(2) Test solution 

(3) Test temperature 

(4) Test aquarium 

(5) Exposure method 

(6) Water for test 

(7) Number offish 

Test water was prepared by dissolving the test substance 

in water for test. 

24.9 ± 0.2°C 

Made of glass, amount of test water; 4 L 

Semi-static method (replacing test water after 48 hours) 

Dechlorinated city water of Saitama 

Analysis of water quality: it was confirmed that water quality 

was adopted to 'the standard of quality of fisheries water' (see, 

Ref.10). 

10 fish/4 L test aquarium 

(8) Experimental period 

(9) Sensitivity test of 

December 20,2010- December 24,2010 

A sensitivity test was conducted using CuS04 • 5H20 as 

Danio rerio 

(10) Test results 

a standard substance. The fish in the same lot which passed in 

the sensitivity test (96h LC50 value; 0.08 to 0.3 mgL-1

) were used 

as 96h LC5o test fish. 

Since the preliminary 96h LC50 value of the test substance 

in Zebrafish (Danio rerio) was greater than 100 mgL-1

, 

the test was done by "a limited test" in which the concentration 

of the test substance was 100 mgL-1

• As a result, it was 

confirmed that the 96h LC50 value was greater than 101 mgL- 1

• 

The concentration was an initial experimental value 

calculated from LC/MS analysis (see, the following table). 

Test water Mortality (%) 

Compound Concentration 

(mgL- 1

) 

Copper (II) sulfate pentahydrate 0.5 

(sensitivity test) 0.05 

Test substance 101 

Control (water for test) -

2.2 Performing a bioconcentration test 

(1) Test fish 

24 48 72 96 

hours hours hours hours 

0 20 70 100 

0 0 0 0 

0 0 0 0 

0 0 0 0 

To confirm the bioconcentration in fish, the carp (Cyprinus ca~pio), representative fish 

species in Japan, was used (SOP 2.1.2) as the test fish for the bioconcentration test. 

Common carp 

Supplier: 

Cyprinus carpio (Lot No. 100823) 

Kitamura Carp Breeding Farm (12-388 Gunchiku, Yatsushiro-shi, 

Kumamoto Prefecture, Japan) 

8 
Date purchased: August 23,2010 

Feeding and acclimatization: 

(2) Feeding 

Kind: 

Manufacturer: 

Feeding method: 

After receiving the fish at the Institute of Ecotoxicology Co., 

Ltd., those with a total length of approx. 8.0 ± 4.0 em were 

selected (SOP 2.4). After the fish were treated with Elevage® 

(4-[2-(5-nitro-2-furanyl)ethenyl]benzoic acid, sodium salt) and 

rock salt, the fish were raised. Fish were selected again on 

January 5, 2011 and 150 fish were transferred to an acclimation 

water-tank. After they were treated again with Elevage®, the 

process of acclimatization was started (SOP 2.3, SOP 2.5). 

Finally, the fish were acclimatized at water temperature 25 ± 

2.0°C for 7 days and it was confirmed that the mortality was 5% 

or less before use in the test. 

Mean body weight 

Mean total length 

3.36 ± 0.21 g 

6.9± 0.2 em 

Mmiality of fish during the experiment 

High exposure level 0% 

Low exposure level 0% 

Control 0% 

(SOP 2.8) 

(SOP 2.8) 

(SOP 2.7) 

Lipid contents (by chloroform/methanol extraction method) 

Before experiment 5.0% (mean value, n=2) (SOP 2.9) 

After experiment 4.8% (mean value, n=2) (SOP 2.9) 

(The conditions ofthe test fish were shown in Ref. 11) 

Mixed feed for carp in pellet form (Swimmy) 

Crude proteins 37.0% 

Crude lipids 2.5% 

Nippon Pet Food Company 

Fish were fed daily in an amount of approximately 1.5% of the 

bodyweight ofthe fish. However, no feed was given on the 

day before sampling of the test fish. 

(3) Performing the test 

Water for test: Same as in Section 2.1 (6) 

Preparation of a stock solution: 

A test substance was added to water for test to prepare 105 mgL"1 

stock solution. The stock solution was diluted with water for test 

to obtain test water. 

High exposure level: 

9 
The stock solution was diluted with water for test to obtain a 

solution ofthe test substance (20 mgL-1

). This was further diluted 

with water for test and continuously flowed through the test 

aquarium. 

Low exposure level: 

Control: 

The stock solution was diluted with water for test to obtain a 

solution of the test substance (2 mgL-1

). This was further diluted 

with water for test and continuously flowed through the test 

aquarium. 

Water for test was continuously flowed through the test aquarium. 

Bioconcentration test equipment: 

see, Ref. I 

Test aquarium: "Flow-through system" (made of glass) 

Capacity 100 L (amount oftest water; 50 L) 

Flow rate 300 mL min·' (432 L day"1

) 

No. 9 aquarium for high exposure level, No. 8 aquarium for low 

exposure level and No. 7 aquarium for the control were used. 

(4) Test concentrations 

The 96h LCso of in Dania rerio was greater than 101 mgL"1 

and the water solubility of was> 1.0 x 105 mgL-1

• The setup 

concentrations for the bioconcentration test were determined to 

be less than the solubility in water. Based on the 96h LC50 value, 

1/505 and 1/5050 were used as safety factors and the test 

concentrations were set as follows. 

High exposure Low exposure Control level level 

Concentration of the test 0.200 mgL-1 0.0200 mgL-1 

substance 

(concentration of 

(5) Test conditions 

Temperature oftest water: 

pH of test water: 

DO of test water: 

(0.193 mgL-1

) 

25.3 ± 0.2°C 

7.5±0.1 

7.4 ± 0.1 mgL-1 

(0.0193 mgL-1

) 

(Temperature, pH and DO of test water were measured on each analysis day, 

see, Ref. 2-2) 

10 

-

-
Measurement of total organic carbon concentrations in the test water (SOP 2.12): 

Light type: 

Photoperiod: 

Experimental period: 

Since solvents or dispersant were not used in this 

test, the total organic carbon concentrations in the 

test water was not measured. 

UV- cut fluorescent lamp 

About 1 0 hours 

January 26, 2011 -February 23, 2011 

The analysis schedule is presented in Section 2.8. 

Number oftest fish: High exposure level 

Low exposure level 

Control 

50 fish 

50 fish 

15 fish 

Number offish analyzed and number repeated: 

2 fish were paired for a single analysis (n=2). 

2 fish were paired for analysis of the control group (n=l). 

11 
2.3 Analyses of fish and test water 

2.3.1 Analytical method for fish (SOP 2.11) 

Pretreatment 

Each six fish in the Concentration Levell and Level 2 and two fish in the control group 

were sampled using a landing net on the analysis day. The fish were temporarily 

paralyzed by placing them in an ice water. The body surface was washed with tap 

water and wiped lightly by gauze. Then, the body weight was measured in a unit of 

0.01 g using a balance (SOP/INS/4.2.2), and the body length from the mouth to the tail 

fin was measured in a unit of 0.1 em. In the test group, two fish were used as one 

sample and analyzed in duplicate, and other sample was put into a plastic bag and 

stored in a freezer after the fish were wrapped with aluminum-foil and packed in a 

plastic bag (SOP 2.15). 

The fish were put into a homogenizer cup after slicing, and acetonitrile (60 mL) was 

added to the homogenizer cup. The mixture was homogenized at 9000 r.p.m. for 6 min. 

(SOP/INS/4.17.2). The lid and the interior ofthe homogenizer cup were rinsed with 

acetonitrile (4 mL) and the washings were transferred to a centrifugal tube. The interior 

of the said homogenizer cup was rinsed three times with 4 mL of acetonitrile and the 

washings were also transferred to the said centrifugal tube. The weight of the 

centrifugal tube was adjusted by adding a small amount of acetonitrile. After 

centrifugal separation for 15 min. (SOP/INS/4.16.3) (9000 r.p.m., at about l0°C), the 

supernatant was filtered into a 100 mL volumetric flask (a glass filter GC50 by 

Advantec MFS, Inc., Tokyo, Japan). The interior ofthe said centrifugal tube and the 

filter paper were rinsed three times with 4 mL of acetonitrile and the washings were also 

filtered into the said volumetric flask. After waiting until the temperature of the solution 

reached room temperature, the volume was adjusted to 100 mL by adding acetonitrile. A 

portion of the constant volume solution was taken with a hole pipette (1 mL) and was 

transferred to a 10 mL volumetric flask. After adding 4 mL of acetonitrile, the volume 

was adjusted to 10 mL by adding ultra pure water. The constant volume solution was 

filtered through a filter (EKICRODISK 13 by Nihon Pall Ltd., Tokyo, Japan). The 

filtrate was analyzed by liquid chromatography mass spectrometer (LC/MS). 

The operations of the pretreatment are shown in the next page as a flow chart. 

12 
I Residue 

Test fish 

(one sample consisting of two fish) 

• Measurements of the total length and bodyweight 

• Slicing 

• Transferred to a homogenizer cup 

~Acetonitrile 60 mL 

• Homogenized (9000 r.p.m., 6 min.) 

~Acetonitrile 4 mL 

(washing the lid and the interior of the homogenizer cup) 

• Transferred to a centrifugal tube 

~Acetonitrile 4 mLx3 (washing the interior of the homogenizer cup) 

• The weight of the centrifugal tube was adjusted by adding a small amount of acetonitrile. 

• Centrifugal separation (9000 r.p.m., at about 10°C, 15 min.) 

I Supernatant I 

• Filtration (glass filter GC50 by Advantec MFS, Inc.) 

(filtered into a 100 mL volumetric flask) 

~Acetonitrile 4 mLx3 

(washing the interior of the centrifugal tube and the filter paper) 

• Waiting until the temperature of the solution reached room temperature 

• Fixed volume 100 mL 

Fixed volume solution 

(acetonitrile) 

~Taken in 1 mL (using a hole pipette) 

• Transferred to a 10 mL volumetric flask 

~Acetonitrile 4 mL 

~Ultra pure water 

• Fixed volume 10 mL 

Fixed volume solution 

(acetonitrile/water(! : 1)) 

• Filtration through a filter (EKICRODISK 13 by Nihon Pall Ltd.) 

Sample for LC/MS analysis 

13 
2.3.2 Analytical method for test water (SOP 2.10) 

Pretreatment 

Test water was sampled (approx. 200 mL from the Concentration Levell and approx. 

200 mL from the Concentration Level 2) from the central area of each aquarium with an 

Erlenmeyer flask. After leaving it standing, the supernatant (1 mL in the Concentration 

Level 1 and 2.5 mL in the Concentration Level 2) was taken using a hole pipette and 

transferred to a volumetric flask (20 mL in Concentration Level 1 and 5 mL in 

Concentration Level2). In Concentration Levell, ultra pure water (9 mL) was added 

to the volumetric flask. The volume was adjusted with acetonitrile to 20 mL in 

Concentration Level 1 and 5 mL in Concentration Level 2. The constant volume 

solution was filtered through a filter (EKICRODISK 13 by Nihon Pall Ltd., Tokyo, 

Japan). The filtrate was analyzed by liquid chromatography mass spectrometer 

(LC/MS). 

The operations of the pretreatment are shown below as a flow chart. 

14 
I 

Concentration Level 1 

Test water I • Sampling water: Concentration Levell, 200 mL; Concentration Level2, 200 mL 

(left standing for a while, waiting for suspended dirt to be deposited) 

• Measurement: Concentration Level 1, 1 mL; Concentration Level 2, 2.5 mL 

(measurement using a hole pipette) 

Taken solution _j 

Concentration Level 2 

• Transferred to a 20 mL volumetric flask 

~Ultra pure water 9 mL 

• Transferred to a 5 mL volumetric flask 

~Acetonitrile 

~Acetonitrile 

• Fixed volume 20 mL 

Fixed volume solution 

(acetonitrile/water(!: 1 )) 

• Filtration through a filter 

(EKICRODISK 13 by Nihon Pall Ltd.) 

Sample for 

LC/MS analysis 

• Fixed volume 5 mL 

Fixed volume solution 

(acetonitrile/water( 1 : 1)) 

• Filtration through a filter 

(EKICRODISK 13 by Nihon Pall Ltd.) 

Sample for 

LC/MS analysis 

The operation of water analysis in the control group (when starting the test) was based 

on the method described in Concentration Level 2. 

15 
2.3.3 Measurement methods 

<Analytical machine> 

Liquid chromatography mass spectrometer (SOP/INS/4.33) 

<Analytical apparatuses and conditions> 

< Liquid chromatography (LC)> 

Analytical apparatus High performance liquid chromatography LC-20AD (Shimadzu Co., Ltd.) 

Column Hypersil GOLD 2.1 mm i.d. x 50 mm (5 J.lm) 

(Lot No. 9966) Thermo Scientific Inc. 

Mobile phase 

Flow rate 

Temperature 

Injection volume 

A: 20 mM ammonium formate/formic acid (1000:1), B: acetonitrile, 

A/B(l:1) 

0.2 mL/min 

40°C 

5 f.lL 

< Mass spectrometer (MS)> 

Analytical apparatus TSQ Quantum Access MAX triple stage quadrupole mass spectrometer 

Ionization method 

Vaporizer temperature 

Spray voltage 

Sheath gas 

AUX gas 

Ion sweep gas 

Capillary temperature 

Scan type 

(Thermo Fisher Scientific Inc.) 

Negative ESI 

450°C 

2500V 

50 

15 

0 

220°C 

Selected ion monitoring (SIM) m/z 180 [M-Lif 

<Quantitative calculation> 

Instrument Xcalibur by Thermo Fisher Scientific Inc. 

Calculation method Two-point calibration method (absolute calibration method) 

The calibration using two standard solutions and the quantification of sample were 

performed on the analysis day. 

2.4 Calibration curve by liquid chromatography mass spectrometry 

2.4.1 Preparation of standard solutions 

The test substances (5.03 mg and 3.15 mg) were weighed using Mettler Toledo 

XS 1 05DU (SOP/INS/4.1.4) into 200 mL and 250 mL volumetric flasks and brought to 

the mark with acetonitrile/water( 1:1 ), respectively. The concentrations of the 

solutions were 25.0242 mgL- 1 and 12.5370 mgL- 1 when considering the purity. The 

concentrations of the solutions were 24.0959 mgL- 1 and 12.0719 mgL-1 when 

16 
converting to the concentration of The solutions were diluted with 

acetonitrile/water( I: 1) to prepare standard solutions for calibration curve. 

2.4.2 Preparation of calibration curve 

The calibration curve was prepared using the standard solutions prepared in Section 

2.4.1 and the linearity was checked. 

Test substance Anion 

Solution concentration concentration Peak area No. 

(mgL"1

) [X] (mgL- 1

) [X] [Y] 

1 0.0050 0.0048 1246726 

2 0.0100 0.0096 2149557 

3 0.0200 0.0193 3499884 

4 0.0501 0.0483 7806108 

5 0.1001 0.0964 15009834 

Calibration curve Y = 149267349.5345 X +616562.7686 

(by the least square method) 

r = 1.000 

[Y] 

c;j 

~ 

~ 8000000 f-------.rat'-+----......J 

ll.) 

Q.. 

0 0. 06 0. 12 

Concentration of (mgL"1

) [X] 

1111 The chromatograms used when preparing the calibration curve are shown in Ref. 6. 

2.5 Recovery test and blank test for 

2.5.1 Recovery test from fish for (SOP 2.11) 

Fish bodies (two fish paired as one sample) were crashed with a blender after mincing, 

added an acetonitrile solution (1 ml) of the test substance , an analytical sample 

was prepared according to Section "2.3.1 Analytical method for fish" and was analyzed 

by liquid chromatography mass spectrometer. The recovery rates of were found to 

be as follows. 

The amount of an addition of the test substance (the amount converted to : 21.9 j..tg) 

corresponded to 3.40 j.lgg·1 relative to the fish 3.22 g (6.44 g/2 fish) and the 

bioconcentration factor in Concentration Level2 (the concentration converted to : 

0.0193 mgL- 1

) was approximately 176-fold. 

Added group 

Fixed Quantity of addition Recovery Recovery Sample Fish weight Dilution 

No. (g) volume factor Measured Calculated measured rate 

(mL) concentrated areas (%) areas (mgL"1

) 

1 3.12 100 10 4025794 0.0219 3951983 98.2 

3.21 

2 3.14 100 10 4025794 0.0219 3953287 98.2 

3.15 

3 3.38 100 10 4025794 0.0219 3849419 95.6 

3.22 

Mean recovery rate: 97.3 ± 1.2% 

17 
<Background of the fish> 

After measuring bodyweight and total length offish bodies (two fish were paired as one 

sample), the fish bodies were crashed with a blender after mincing. After adding 1 mL of 

acetonitrile, an analytical sample was prepared according to Section "2.3.1 Analytical 

method for fish" and was analyzed by liquid chromatography mass spectrometer. No 

background peaks were detected at the position of the peak of 

Control group 

Sample Fish Fixed Measured bodyweight volume Dilution factor No. (g) (mL) areas 

1 3.21 100 10 N.D. 

3.29 

2 3.25 100 10 N.D. 

3.22 

3 3.09 100 10 N.D. 

3.41 

• The chromatograms used when calculating the recovery rates of 

bodies are shown in Ref. 7-1. 

from the fish 

2.5.2 Recovery test from test water for (SOP 2.10) 

Water used for raising fish was sampled and a solution containing the test substance 

was added. According to Section "2.3.2 Analytical method for test water", an 

analytical sample was prepared and analyzed by LC/MS. As shown in the following 

table, the mean recovery rates of were 96.2 ± 1.1% in Concentration Level 1 and 

95.5 ± 1.2% in Concentration Level2. 

Recovery rates in Concentration Level 1 (Added group) 

Amount of Fixed Quantity of addition Recovery Recovery Sample water Calculated 

No. measured volume Measured concentrated measured rate 

(mL) (mL) areas (mgL-I) areas (%) 

I 1 20 2377235 0.0114 2270545 95.5 

2 1 20 2377235 0.0114 2266589 95.3 

3 1 20 2377235 0.0114 2321802 97.7 

Mean recovery rate: 96.2 ± 1.1% 

18 
Recovery rates in Concentration Level 2 (Added group) 

Amount of Fixed Quantity of addition Recovery Recovery Sample water Calculated 

No. measured volume Measured concentrated measured rate 

(mL) (mL) areas (mgL-1) areas (%) 

1 2.5 5 2413659 0,0118 2334903 96.7 

2 2.5 5 2413659 0.0118 2315855 95.9 

3 2.5 5 2413659 0.0118 2264899 93.8 

Mean recovery rate: 95.5 ± 1.2% 

<Background of the test water> 

Water used for raising fish was sampled and ultra pure water was added. According to 

Section "2.3.2 Analytical method for test water", an analytical sample was prepared and 

analyzed by LC/MS. No background peaks were detected at the position of the peak of 

either in Concentration Level 1 or in Concentration Level 2. 

c ontro group 

Concentration Level 1 Concentration Level 2 

Amount of Fixed Amount of Fixed Sample water volume Measured Sample water volume Measured 

No. measured (mL) areas No. measured (mL) areas 

(mL) (mL) 

1 1 20 N.D. 1 2.5 5 N.D. 

2 1 20 N.D. 2 2.5 5 N.D. 

3 1 20 N.D. 3 2.5 5 N.D. 

111 The chromatograms used when calculating the recovery rates of 

shown in Ref. 7-2. 

from water are 

2.6 Quantifiable limit 

2.6.1 Quantifiable limit for fish 

In the LC/MS chromatograms, there were no background peaks found due to fish body 

at the position of . If the peak area of 800000 counts is set as a detectable limit, the 

quantifiable bioconcentration factor is shown in the following table. 

Setup concentration in Detectable Quantifiable Bioconcentration factor-'' 

each concentration level concentration 1 l limit2

l Concentration Concentration 

(concentration of (mgL-1

) (IJ.gg-1) Levell Level2 I (0.193 mgL-1

) (0.0193 mgL-1

) 

Concentraion Level 1 

0.193 mgL-1 

0.0012 0.192 1-fold or greater 10-fold or greater Concentraion Level 2 

0.0193 mgL-1 

(Note I) The detectable concentration was calculated from the calibration curve in Section 2.4.2. 

(Note 2) The quantifiable limit in the fish bodies was calculated by the calculation formula shown in the next page. 

19 
The quantifiable limit was calculated as the fish bodyweight; 6.44 g (3.22 g per fish, 2 fish 

were paired as one sample), recovery rate; 97.3%, fixed volume; 100 ml and dilution factor; 10. 

Quantifiable limit (f,!gg·I) = Detectable concentration (mgL"1

) x Fixed volume (mL) x Dilution factor x 100 

Recovery rate(%) x Fish bodyweight (g) 

(Note 3) 

Quantifiable limit (ugg·1

) Bioconcentration factor (BCF) = Setup concentration in the test group (theoretical value) (mgL"1

) 

2.6.2 Quantifiable limit for test water 

In the LC/MS chromatograms, there were no background peaks observed due to test 

water at the position ofthe peak of . Ifthe peak area of800000 counts is set as a 

detectable limit, the quantifiable limit for test water is shown in the following table. 

Setup concentration in each 

concentration level Detectable Quantifiable limit2

l (mgL"1

) 

(concentration of concentration1

l 

Concentration Concentration (mgL"I) Concentration Concentration 

Levell Level2 Levell Level2 (0.193 mgL"1

) (0.0193 mgL"1) 

0.193 mgL"1 0.0193 mgL"1 0.0012 0.0249 0.00251 

(Note 1) The detectable concentration was calculated from the calibration curve in Section 2.4.2. 

(Note 2) The quantifiable limit for test water was calculated by the following calculation formula. 

The quantifiable limit was calculated as the fixed volume; 20 mL in Levell and 5 mL in Level 2, 

recovery rate; 96.2% in Levell and 95.5% in Level 2 and amount of water measured; 1 mL in 

Level 1 and 2.5 mL in Level 2. 

Quantifiable limit (mgL"1

) = 

2.7 Preliminary test 

Detectable concentration (mgL"1

) x Fixed volume (mL) 

Recovery rate(%) x Amount of water measured (mL) X 100 

Test fish (Cyprinus carpio) were exposed to test water (the concentration converted to 

0.0193 mgL"1

) in Concentration Level2 (the flow-through system). On the ih day 

of exposure, the test fish (2 fish as a pair) and the test water were sampled and analyzed 

according to Sections "2.3.1 Analytical method for fish" and "2.3.2 Analytical method 

for test water". As a result, the BCF of was found to be under a detectable limit 

and less than 8-fold. 

20 
2.8 Schedule of analyses of fish and test water 

Analyses of test water and fish were conducted according to the following schedule: 

Number of 

Sampling date days after Test group Control group 

exposure 

Analysis of water 

Exposure January 26,2011 DayO Analysis of water Analysis of fish 

period Lipid contents 

(28 days) February 2, 2011 7th day Analysis of water -

Analysis of fish 

February 9, 2011 14th day Analysis of water -

Analysis of fish 

February 16, 2011 21 51 day Analysis of water - Analysis of fish 

February 18, 2011 23rd day Analysis ofwater - Analysis of fish 

February 23,2011 28th day Analysis of water Analysis of fish 

Analysis offish Lipid contents 

On the day of analysis ofwater, pH, DO (adjusted to be approximately 7 mgL-1

) and 

water temperature were measured for each aquarium and the data were recorded (pH 

meter SOP/INS/4.3.3, DO meter SOP/INS/4.4.4). 

In compliance with SOP 2.10 (measurement of concentrations of the test substance in 

the test water), analysis of test water was conducted according to Section "2.3.2 

Analytical method for test water". The changes in the concentration of the test 

substance in water were confirmed to be ±20% or less relative to the mean of the 

measured values during the exposure period. 

In compliance with SOP 2.11 (measurement of concentrations of the test substance in 

the fish), analysis offish was conducted according to Section "2.3.1 Analytical method 

for fish". In compliance with SOP 2.9 (measurement of lipid contents in the fish), lipid 

contents were measured according to the "chloroform/methanol extraction method". 

The changes in the lipid contents from the beginning ofthe experiment to the end of the 

experiment were confirmed to be ± 25% or less. 

Storage before analysis offish and treatment ofwaste during analysis offish were 

performed in compliance with SOP 2.15 (storage before analysis offish) and SOP 2.16 

(discharging waste during analysis offish). 

21 
3. Test results 

3.1 Concentrations of in test water 

The concentrations of in the test water (mgL- 1

) are shown in the following table 

with respect to the concentration levels. 

~ e DayO 7th day 14th day 2151 day 23rd day 28th day 

p 

Concentration Level I 0.177 0.181 0.177 0.181 0.187 0.185 

Concentration Level 2 0.0186 O.oi88 0.0184 0.0184 0.0193 0.0188 

Umt. mgL ·l 

• Analytical data during the analysis oftest water are shown in Ref.5. 

Based on the results shown above, the mean concentration during the test period and the 

standard deviation were as follows. 

Concentration Level 1: 0.181 ± 0.00373 mgL -I 

Concentration Level2: 0.0187 ± 0.000308 mgL"1 

The following calculation formula was used. 

Mean Standard deviation 

11 

L.>i 2> ~=i::!____=- n n 

111 Changes over time in the concentrations of the test substance in water, water 

temperature, pH and DO are shown in Ref. 2. The chromatograms at the time of 

analysis on Day 0, 7, 14, 21, 23 and 28 after exposure are shown in Ref. 9-1. Further, 

the chromatograms at the time of analysis of water in the control group are shown in 

Ref. 9-2. 

22 
3.2 Bioconcentration factors (BCFs) 

According to the results of the bioconcentration test, the bioconcentration factors (BCFs) 

of were under a detectable limit in both Concentration Levels (less than 1-fold in 

Concentration Level 1 and less than 11-fold in Concentration Level 2). Since it was 

confirmed that the steady-state was achieved within 28 days in both Concentration 

Levels, exposure was finished. The bioconcentration factors (BCFs) of are shown 

below with reference to the day of analysis. 

Number of Concentration Level 1 Concentration Level 2 

days after Sample No. Bioconcentration Sample No. Bioconcentration 

exposure (n==2) factor (BCF) (n==2) factor (BCF) (Days) 

7 No.1 <1 No.1 <10 

No.2 <1 No.2 < 10 

14 No.1 <1 No. I <11 

No.2 <1 No.2 <10 

21 No.1 <I No. I < 10 

No.2 <1 No.2 < 10 

23 No.1 <1 No. I < 10 

No.2 <1 No.2 < IO 

28 No.1 <1 No.1 < 10 

No.2 <1 No.2 < 10 

(Note 1) The sample numbers in Concentration Levels 1 and 2 are symbols given to the fish taken from 

each concentration group when the amount of accumulation was measured. 

(Note 2) The fish bodyweight during the analysis and other analytical data are shown in Ref. 4, and 

the chromatograms obtained at the time of analysis on the 7fu, 14th, 21 5

\ 23rdand 28th day are 

shown in Ref. 8-1. The chromatograms obtained at the time of analysis of fish in the control 

group are shown in Ref. 8-2. 

(Note 3) The time-dependent changes in the bioconcentration factors in fish (BCF) are shown in 

Ref. 3. 

(1) Mean ofbioconcentration factors (BCFs) and standard deviation 

The mean of the bioconcentration factors (BCFs) of during the experiment period 

and the standard deviation are shown below. 

Concentration Level 1 BCF =The mean of the BCFs was not calculated because 

all the BCFs were less than the detectable limit. 

Concentration Level2 BCF =The mean of the BCFs was not calculated because 

all the BCFs were less than the detectable limit. 

Bioconcentration factor (BCF) == Concentration of 

in the aquarium (mgL"1 Concentration of ) 

Mean Standard deviation 

n 2>; 2> 

;: == .!.::!.__ == --

n n n n 

23 
(2) Confirmation of a steady-state 

Since the bioconcentration factors (BCFs) of in both Concentration Levels were 

less than 50-fold during the 28-day exposure period, the steady state was assumed to be 

achieved on the 28th day of exposure and exposure was completed on the 28th day. 

< Reference items> 

Test fish in the control group were analyzed before exposure Day 0 (n=1) and at the end 

of exposure Day 28 (n=1). No background peaks due to fish were detected in the LC/MS 

chromatograms at the position of the peak of as in the control group in the added 

recovery test. These chromatograms are shown in Ref. 8-2. 

3.3 Evaluation, discussion of the results and conclusion 

The test substance is a salt consisting of . In this test, the bioconcentration of 

only was confirmed. Since lithium bromide [1-110], listed as an existing chemical 

substance, is not high bioconcentration, the bioconcentration ofLi is considered to be 

confirmed by analogy. 

The 96h LC5o value of the test substance in Zebrafish (Dania rerio) was greater 

than 101 mgL-1

• Since the solubility in water was >1.0 xl05 mgL- 1, two concentrations 

of the test substance in water were set up to be 0.2 mgL-1 ( : 0.193 mgL-1

) for the high 

exposure level (Concentration Levell) and 0.02 mgL-1 ( : 0.0193 mgL-1

) for the low 

exposure level (Concentration Level2). According to the results of the bioconcentration 

test, the bioconcentration factors (BCFs) in both Concentration Levels 1 and 2 were less 

than 50-fold. Thus, a steady state was assumed to be achieved within 28 days of exposure. 

It was confirmed that the bioconcentration was under a detectable limit and the 

bioconcentration factors were less than 1-fold in Concentration Levell and less than 11-

fold in Concentration Level 2. Therefore, bioconcentration of 

of the test substance was confirmed to be low. 

24 
Table of contents for references 

Page 

Reference 1 Bioconcentration test equipment outline 26 

Reference 2 Aquatic environment (concentrations, water temperature, pH and DO) 27 

2-1 Concentrations of in water 27 

2-2 Measurement oftemperature, pH and DO in water 27 

Reference 3 Time-dependent changes in bioconcentration factors 28 

Reference 4 Fish analysis data 29 

Reference 5 Water analysis data 32 

Reference 6 Chromatograms for the preparation of calibration curve 35 

Reference 7 Chromatograms of for recovery rate 38 

7-1 Chromatograms of for recovery rate from fish 38 

7-2 Chromatograms of for recovery rate from water 43 

Reference 8 Chromatograms in fish 52 

8-1 Test group 52 

8-2 Control group 67 

Reference 9 Chromatograms used when measuring the concentrations in water 70 

9-1 Test group 70 

9-2 Control group 82 

Reference 10 Water quality test 84 

Reference 11 Records of observations of the conditions of the test fish (carp) 86 

Reference 12 IR spectrum for the test substance 88 

25 
Reference 1 Bioconcentration test equipment outline 

A 

B 

A: Tap water 

B: Filter/activated charcoal tank 

(dechlorination equipment) 

C: Tank for water for test 

(dechlorinated water) 

E 

G .............................................................................. 

... ....................................................................... .. . .. ........................................................................ ... . ... ........................................................................ .. . ... ........................................................................ .. . ... ······································································· .. . . ,.. .. . ~11 ~11111~~1~j~1111111j1111111jjjjj~1~1l111111111~jjj~jjjjj1jjj~jj 1~j 

m 111~~~1111111~11jj~jjjj1j1mmmlml~H~11111mm1l m 

11U~!~~~~~~~~~~~~lmrrrmmmmmm11l11llll1Wl111111llll!~~~!~Ull 

E: Tank for stock solution of test substance 

(lOL) 

F: Mixing tank (2L) (mechanical stirrer) 

G: Test aquarium (lOOL) 

G-1: Aeration 

D: Regulator for controlling the flow rate and 

mixing ratio 

G-2: Temperature regulator/heater 

D-1 : Water for test 

D-2: Stock solution of test substance 

26 
Reference 2 Aquatic environment (concentrations, water temperature, pH and DO) 

2-1 Concentrations of in water 

+ Concentration Level 1 Group Ill Concentration Level 2 Group 

:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: 

""""""""""""""""""""""""""""""".""""""""""""""""""""""""""""""".""""""""""""""""""""""""""""""" .. """""""$""""""""""""""""""""" 

0. 1 :::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: 

----------------------------------------------------------------------------------------------------------------------------------

11'"""""""""""""""""""""""""""""""1111""""""""""""""""""""""""""""""11111""""""""""""""""""""""""""""""1!1"""""""1111"""""""""""""""""""""1~ 

0. 01 1;::=::.=·::=::=::=::=:::=::=::=::=::·=.::=::=::=::=::=:::=::=::=::=:::=::=::=::=::=:::=::""::=::=:::=::=::=::=::=:::=::=::=::=:::=::=::=::=::=:::=::=::=::=::=:::=::=::=::=:::=::=::=::=::=:::=1:: 

---------------------------------------------------------------------------------------------------------------------------------- ---------------------------------------------------------------------------------------------------------------------------------- ----------------------------------------------------------------------------------------------------------------------------------

0.001 ~------------~------------~--------------~------------~ 

Number of 

days after 

0 7 14 21 28 

Time after exposure (days) 

2-2 Measurement of temperature, pH and DO in water 

The aquatic environment was measured at a fixed time (10 a.m.). For pH, 

water was sampled from the center of the aquarium and measured with a 

pH meter (SOP/INS/4.3.3). Further, DO in water was measured at the 

center of the aquarium using a DO meter (SOP/INS/4.4.4). The results are 

shown in the following table. 

Concentration Level 1 Concentration Level 2 Control group 

Water DO Water DO Water DO 

exposure temperature pH (mgL"1

) temperature pH (mgL"1) temperature pH (mgL"1

) (OC) CCC) CCC) 

0 

7 

14 

21 

23 

28 

24.8 7.4 7.5 25.1 7.4 7.5 25.5 

25.4 7.4 7.5 25.2 7.5 7.3 25.3 

25.5 7.5 7.4 25.5 7.5 7.4 25.4 

25.1 7.5 7.5 25.4 7.4 7.4 25.1 

25.4 7.6 7.5 25.4 7.6 7.4 25.4 

25.2 7.6 7.4 25.1 7.5 7.5 25.3 

[DO in water] 

+ Concentration Level 1 Ill Concentration Level 2 .A Control Group 

10 

9 

~ 8 

5 

8 7 

6 

5 

a 

0 7 

Ill lfll lfll ~ 

14 21 28 

Time after exposure (days) 

27 

7.4 7.3 

7.5 7.3 

7.5 7.5 

7.4 7.4 

7.6 7.4 

7.6 7.5 
Reference 3 Time-dependent changes in bioconcentration factors 

[Concentration Level 1] 

..!- indicates less than a detectable limit. 

100 ::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: 

----------------------------------------................................................................................................................................................................. .. """" (.;.. 

u 

6 

.... 

..9 10 u 

..::: 

::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: ---------------------------------................................................................................................................................................................................. .. !::: 

0 

·~ 

!::: 

0 

u 

!::: 

0 

u 

0 

co 

0.1 

0 7 14 21 28 

Number of days after exposure (days) 

[Concentration Level 2] 

..!- indicates less than a detectable limit. 

100 

G:' 

u 

6 

.... 10 ..9 

u 

..::: 

!::: 

.g 

0:: 

~ 0 

u :::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: 

............................................................................................................................................................................................................................................................... 

---------------------------------------------------------------------................................................................................................... .. :::: ------------------------------------------------------------------------------------------------------------------ 0 

u 

0 

co 

0.1 

0 7 14 21 28 

Number of days after exposure (days) 

28 
N 

\0 

Reference 4 Fish analysis data 

Fish analysis data (Concentration Level 1) 

Items I A B c D E I F I G I H I I I J I K 

Fixed Concentration Amount volume Dilution calculated by two- deposited in Fish Recovery rate 

Concentration Concentration Bioconcentration 

1 

D f 

of the Peak area point calibration weight in fish body in water tank ateo 

factor fish body (%) factor 

Samples ""- !solution curve (J.tg) (g) (J.tgg-1) (mgL-1) (BCF) measurement 

(mL) (mgL-1) 

Day 7No.1 I 100 10 N.D. <0.0012 <1.2 I ~.69 [3.24, 3.45] I 97.3 I <0.184 I 0.181 I <1 I 2011.2.2 

.... P..~~-.?. .. ~.?.:.~---······ ...... ~_gg ______ ................. !.?. ....................... ~:P.: .................... ::':?..:.~9. .. ~-~ ....................... :!.:.~ .............. ?.:.~?. .. P.:.~~:. .. ~.:.?.?.J.. ............. ?..?.:? ....................... ~?..:} .. ~-~ .................. ~.:.!.~.! ............................. ::':L ................... ~-~}--~ .. : .. ~.: .... ~ .. . 

Day 14 No.I 100 10 N.D. <0.0012 <1.2 6.87 [3.58, 3.29] 97.3 <0.180 0.177 <1 2011.2. 9 

... P.~~-J-~ .. ~~-:~ ..... .... } ..~~------ .............. J9. ....................... ~:P.: .................... ::':?..:.~~--~-~ ........................ ~.!..:.~ .............. ?.:.?.?. .. E:.!}.:. .. ~.:.?.~J.. ............ .?..?.:.~ ....................... ::':?..:.!. .. ~-~--------· ......... ~.:.!.?..?. ........................... :::L .............. ----~g-~ .. !.: ... ~.: .... ?. .. . Day 21 No.I 100 10 N.D. <0.0012 <1.2 6.83 [3.41, 3.42] 97.3 <0.181 0.181 <1 2011. 2.16 

.. P.~? .. ~L-~.?..:~ ...... ... JQg ...... ................ !..~ ...................... ~.:.P..: .................... ::':~.:-~~.!.-~ ....................... ::':.!..:.~ ............. ~.:-~~ .. P.:~.?..~.-~.:-~ .. ~L ............. ?.?..:.~ ....................... :::~.:.!..~.! ................... ~.:.!..~.!. ............................ ::':.~ ..................... ~-~-~).: ... ~.:-~ .. ?. .. . Day23No.1 100 10 N.D. <0.0012 <1.2 6.62[3.14,3.48] 97.3 <0.186 0.187 <1 2011.2.18 

... P.~t~.~---~~-:~ ..... ..... L~~-----· ................. ~-~ ....................... ~:P.: .................... ::':?..:.~~--~-~ ........................ ~.!.:.~ .............. ?.:.?..?. ... P.:.~?.-~ .. ~.:~~1. ............ .?..?.:.~ ...................... ::':?..: .. ~ .. ~-~ .................. ~.:.!..~.?. ........................... ::':L ................... ~.~-~ .. !..: ... ~.:-~-~ .. . 

Day 28 No.1 100 10 N.D. <0.0012 <1.2 6.91 (3.11, 3.80] 97.3 <0.178 0.185 <1 2011.2.23 

Day 28 No.2 100 10 N.D. <0.0012 <1.2 

Calculation methods: Amount deposited in fish 

Concentration in fish 

B ioeoneentration factor (BCF) 

6.81 (3.53, 3.28] 

E =AxBxD 

H= EX 100 

FxG 

J = I 

97.3 <0.181 0.185 <I 2011.2.23 

Total value of2 fish was used for F. 

(Note) Sample No. is a symbol given when two fish were paired when sampling fish and analyzed at n=2. 

For the samples showing less than the detectable limit (peak area 800000), N.D. is recorded in the column C and the calculation was made by 

replacing the detectable limit of0.0012 mgL-1 in the aforementioned D. 
w 

0 

Fish analysis data (Concentration Level 2) 

Items I A B c D E I F I G I H I I I J I K 

Fixed Concentration Amount volume Dilution calculated by two- deposited in Fish 

Recovery rate 

Concentration Concentration Bioconcentration 

1 

D . f 

of the Peak area point calibration weight in fish body in water tank ateo factor 

"'- !solution 

factor fish body (g) (%) (flgg-1) (mgL"1

) (BCF) measurement 

Samples curve (f.! g) (mL) (mgL"1

) 

Day 7No.1 I 100 10 N.D. <0.0012 <1.2 6.76 [3.58, 3.18] I 97.3 I <0.1 82 I 0.0188 I <10 I 2011.2. 2 

... P.~~-.?. .. ~.?.:.~ ......... ···---~--~~---··· ················-~-~---··--········· ....... ~.:P.: .................... ::~.:-~~--~-~---········· ........... :.!.:.~ .......... .... ~:-~~ ... P.:.~.?..~.-~.:.?..~1 ............. ?.?:.~ ...................... ::~.: .. ~.-~g········· ......... ~.:-~} .. ~~---······ .............. :.!.~ .............. ··---~-~-~--~-: ... ~.: .... ~---

Day14No.l 100 10 N.D. <0.0012 <1.2 6.25[2.89,3.36] 97.3 <0.197 0.0184 <11 2011.2.9 

·---~-~~-}-~ .. ~-?..:.~ ......... J.~~------ ................. .~ ................ ....... ~.:P.: .................... ::~.:.?.~ .. ~-~ ............. ........... :::~.:-~ .......... .... ~.:.?. .. ~ .... P.:.~.?.:. .. ~.:-~.?..1.. ............. ?..?.:.~ ............. ......... ::.~:} .. ~.?. .................. ~.:-~.~--~~ ........................ :::~-~ .................. ~.?.-~ .. ~ .. : .. ~.: .... ?. .. . Day 21 No.I 100 10 N.D. <0.0012 <1.2 6.89 [3.24, 3.65] 97.3 <0.179 0.0184 <10 2011. 2.16 

----~-~x .. ~~---~-?..:~ ..... L. ... ~ .. ~~--.. ..I. ............. J~ ................ L.. .... ~.:P.: ...... ..I. ........... ::~.:.?.~ .. ~-~------ ...... .I.. ......... :::~.:~ .......... L..?..:.?..~ .. E:.?..?.:. .. ~.:.?..~.LI... .......... ?..?.:.~ ........... ..I. ........ ::.?.:} .. ~-~---- .... .l. ........ ~.:-~-~--~~ ......... .1.. ............ :::~.~--------------L-~-~-~---~ .. : .. ~.:.~ .. ?. .. . 

Day 23 No.I 100 10 N.D. <0.0012 <1.2 

Day 28 No.2 100 10 N.D. <0.0012 <1.2 

Calculation methods: Amount deposited in fish 

Concentration in fish 

Bioconccntration factor (BCF) 

6.49 (3.29, 3.20] 

6.76 (3.42, 3.34] 

E=AxBxD 

H= EX 100 

FxG 

J = _lL 

I 

97.3 <0.190 0.0193 <10 2011. 2.18 

97.3 <0.182 0.0188 <10 201 I. 2.23 

Total value of2 fish was used for F. 

(Note) Sample No. is a symbol given when two fish were paired when sampling fish and analyzed at n=2. 

For the samples showing less than the detectable limit (peak area 800000), N.D. is recorded in the column C and the calculation was made by 

replacing the detectable limit of0.0012 mgL·' in the aforementioned D. 
w 

....... 

Fish 

Items 

Samples 

DayO 

---------------------

Day28 

. d (C 

' 

A 

Fixed 

volume 

of the 

solution 

(mL) 

100 

------------------

100 

) 

/ 

B c D 

Concentration 

calculated by 

Dilution Peak two-point 

factor area calibration 

curve 

(mgL-1

) 

10 N.D. -

------------------------- ------------- -----------------------------

10 N.D. -

E F G H I J K 

Amount 

deposited Fish Recovery Concentration Concentration Bioconcentration Date of in fish weight rate in fish body in water tank factor 

body (g) (%) (!lgg-') (mgL-1

) (BCF) measurement 

(!lg) 

- 6.84 [3.53, 3.31] - - - - 2011. 1.26 

------------------- --------------------- ------------------- ------------------------- -------------------------- ------------------------- ----------------

- 6.62 [3.66, 2.96] - - - - 2011. 2.23 
w 

N 

Reference 5 Water analysis data 

Water analysis data (Concentration Level I) 

Items A B c D E 

Fixed Concentration Absolute amount Amount of volume calculated by in the fixed water of the Peak area two-point 

solution calibration curve volume solution measured 

Samples (mL) (mgL-1

) (f!g) (mL) 

DayO 20 1873661 0.0085 0.170 1 

Day7 20 1921305 0.0087 0.174 1 

Day 14 20 1895936 0.0085 0.170 1 

Day21 20 1931160 0.0087 0.174 1 

Day23 20 1967483 0.0090 0.180 1 

Day28 20 1964054 0.0089 0.178 1 

Calculation methods: Absolute amount in the fixed volume solution 

Concentration in the aquarium 

F 

Recovery rate 

(%) 

96.2 

96.2 

96.2 

96.2 

96.2 

96.2 

D = AxC 

G = __ D X lQO_ 

ExF 

G H 

Concentration Date of in water tank 

(mgL-1

) 

measurement 

0.177 2011. 1.26 

0.181 2011.2. 2 

0.177 2011.2. 9 

0.181 2011.2.16 

0.187 2011. 2.18 

0.185 2011. 2.23 
w 

w 

Wat · data (C tration L 12) ' / 

Items A B C D E F G H 

Fixed Concentration Ab 1 t A f 

so u e amount mount o . volume calculated by . Concentration 

fth P k t 

. t m the fixed water Recovery rate . k Date of 

o e ea area wo-pom . 0 m water tan 

I . J"b . volume solutiOn measured (Yo) ( L.1) measurement 

so utJOn ca 1 rat10~ curve (~g) (mL) mg 

Samples (mL) (mgL ) 

Day 0 5 1937105 0.0089 0.0445 2.5 95.5 0.0186 2011. 1.26 

Day 7 5 1970474 0.0090 0.0450 2.5 95.5 0.0188 2011.2. 2 

Day 14 5 1941077 0.0088 0.0440 2.5 95.5 0.0184 2011.2. 9 

Day21 5 1943578 0.0088 0.0440 2.5 95.5 0.0184 2011.2.16 

Day23 5 1988219 0.0092 0.0460 2.5 95.5 0.0193 2011.2.18 

... Day 2~ _5_ 198577~L_ 0.0090 0.0450 2.5 95.5 0.0188 2011.2.23 

Calculation methods: Absolute amount in the fixed volume solution 

Concentration in the aquarium 

D = AxC 

G = _I)__2_!QQ_ 

ExF 
w 

..p. 

w 

Items 

Samples 

DayO 

· data (C 

' 

A 

Fixed 

volume 

of the 

solution 

(mL) 

5 

) 

/ 

B c 

Concentration 

calculated by 

Peak area two-point 

calibration curve 

(mgL-1

) 

N.D. -

D E F G H 

Absolute amount Amount of 

in the fixed water Recovery rate 

Concentration Date of in water tank volume solution measured (%) (mgL-1

) 

measurement 

(!!g) (mL) 

- 2.5 - - 2011. 1.26 
Reference 6 Chromatograms for the preparation of calibration curve 

Calibration curve 

Standard solution 

0.0048 mgL"1 

Calibration curve 

Standard solution 

0.0096 mgL"1 

Dafafile: C186CALS1·2 Acquisition Date: 

Original Oala Path: C:\XCAliBUR\DATA\KANEKOIC Samplo!O: 

186\CAL Sampla Name: Sid. 0.0046 mg/1.. Comments: 

Sample Type: Std Bracket Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE : SHIMADZU LC-20AD COLUMN 

ELUENT : Hyparsil GOLD 2.1 x 50 mm (5 um) : A; 20 mM Amrrumlum !orrnale aqJformlc acid (1000:1) 

B; Acefonlttife 

FLOW RATE COLUMN TEMP INJECT VOL 

NB(1:1) 

: 0.2mllmln 

: 313K : 5uL 

"' ANALYTICAL CONDITIONS (MS) "' 

MS 1YPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILlARY TEMP : 493 K SCAN TYPE : SIM (mlz 160) 

Data File: C186CALS2·2 Acqulslllon Oata:-

Or!glnal Oala Path: fJ)l(~IBUR\DATA\KANEl(O\C Sample!D: 

Sampla Namo: Std. 0.0096 mgt1.. Comments: Samplo Typo: Sid Bn>cket Operator: 

................................................................................................................................... "' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC-20AD COLUMN : Hyporsll GOlD 2.1 x 50 mm (5 um) 

ELUENT : A: 20 mM Ammonium formate aqJformfc o<'.d (1000:1) B; Acetonllrlle AIB(1:1) FlOW RATE : 0.2 mllmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"' ANALYTICAL CONDITIONS {MS) -

MS TYPE : Thermo Ftsher SclenUrre TSQ Quantum Access MAX IONIZATION : Nagalive ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM {m/z 1 80) 

................................................................................................................................... 

35 
Calibration curve 

Standard solution 

0.0193 mgL-1 

Calibration curve 

Standard solution 

0.0483 mgL-1 

Data Fife: CfB6CAlS3·2 Acquisition Date: Original Data Path: C:\XCAUBUR\DATA\KANEKO\C SampleiD: 186\CAL Sampto Name: Ski. 0.0193 mgn. Commenfs: 

Sample Type: Sid Bracket Oporator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

H?LC TYPE : SHIMADZU Lc-20AD COLUMN : HyporsiiGOLD 2.1x50mm(5um) ELUENT : A; 20 mM Alnrn<><'Jum fomtale aq.llormlc acid (1000:1) 

B; Acetcnlftl!e NS(f:f) FLOW RATE : 0.2 mVmin COLUMN TEMP : 313 K INJECT VOL : 5 ul 

'" ANALYTICAL CO!IDITIONS (MS) "' MS TYPE : Thermo Fisher Sc!an!ltic TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPAAYVOLTAGE : 2500V 

SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

1\T;O.CG·~OO 

-,._, 

lto:«rl 

-::= ·-

Data Fifo: Cf86CAlS7·2 Acqulsnlon Oate: Original Data Path: C:IXCAUBUR\OATA\KANEKO\C SampleiO: 186\CAL Sample Namo: Std. 0.0483 mgll Comments: 

Sample Type: Sid Bracket Operator: 

"' AIIAL YTICAL CONDITIONS (HPLC) "' 

HPlCTYPE : SHIMADZU LC.20AD COLUMN 

ElUENT : Hypersll GOlD 2.1 x EO mm (5 um) 

A; 20 mM Ammonium fo111U!Ie •qJformlc add (1000:1) Bi Ac:e!onitrile 

FLOW RATE COLUMN TEMP INJECT VOL 

NS(1:1) : 0.2mUmln : 313K : 5ul 

'" ANALYTICAL CONDITIONS {MS) '" 

MS TYPE : Thermo Fisher Sclenllfic TSQ Quantum Acc.,s MAX IONIZATION : Negallve ESI 

VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : EO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARYTEMP : 493 K SCANTYPE : SIM{mlz180) 

'""" 

"''"" Jt~~ 

..,., '''"' 

.....,· 

..,... 0· 

0.0

36 
Calibration curve 

Standard solution 

0.0964 mgL·1 

Calibration curve 

Chromatogram of 

solvent for adjustment 

Data File: C166CA!.S4·2 Acqulsllfon Dalo: 

Original Data Path: 0:\XCAUBURIOATA\KANEKO\C SampleiO: 

186\CAL 

Sample Name: Std. 0.0964 mg/L Comments: 

Sampltt Type: Std Bracket Operator: 

'" ANAl \'TICAl CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LG-20AD COI.UMN Hypersl GOI.O 2.1x 50 mm (5 urn) ELUENT A; 20 mM Ammonium lormale aq.lfotmle aeld (1000:1) 

B;AcetonlkHs 

A/8(1:1) FLOWRATE : 0.2mlhnln 

COLUMN TEMP : 313 K INJECTVOL : 5uL 

·~ ANALYTICALCONDinONS(MS) •·• 

MS TYPE : Thermo Fisher Scientific TSQ Quanlum Access MAX 

IONIZATION : Negallve ESI VAPORIZER TEI.lP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Data Fils: C1 86CALS0·1 Acquisition Date: 

Original Data Path: C:IXCALIBUR\OATA\KANEKO\C SamplaiD: 

186\CAL 

Sample Name: Solvent Comments: 

Sampte Type: Blank Operator: 

"'ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE : SHIMAOZU LG-20AO COI.UMN 

ELUENT : Hypersll GOL!l 2.1x so mm (5 urn) : A; 20 mM Ammonium lormato eqJiorm!c acid (1000:1) 

B;Aeelonll!lle 

FLOW RATE COLUMN TEMP INJECT VOL 

A/8(1:1) : 0.2mllmln 

: 313K : SuL 

"' ANAL\'TICALCOIIDITIOilS (MS) "' 

MS TYPE : Thermo Asher Scitnlifi-c TSQ Qua."'otum Access: ~tAX 

IONIZATION : Negalive ESI VAPORIZER TEI.IP : 723 K SPRAYVOLTAGE : 2500V 

SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

__

37 
Reference 7 Chromatograms of for recovery rate 

7-1 Chromatograms of for recovery rate from fish 

Recovery test 

(Fish analysis) 

Standard solution 

0.0048 mgL·1 

Recovery test 

(Fish analysis) 

Standard solution 

0.0483 mgL·1 

DalaF!Ie: 

Original Data Path: 

Sample Name: 

Sample Type: 

C1B8FRS1 Acquisition Dale: 

C:IXCALIBUR\OATA\kaneko\C18 SampleiD: 

6\FR 

Sld. 0.0048 mgll Comments: 

Sid Bracket Opor.ator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC-2!lAO COlUMN : Hypersll GOLO 2.1 x50 mm (5 um) ElUENT : A; 20 mMAmmonlum formate aqJf01mk: acid (1000:1) 

B; Acelonllli!e 

NB(I:1) FLOW RATE : 0.2mllmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Tharmo'Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negall\'0 ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

• ..'1-·

Oafa FUe: C1e6FRS2·1 AcquisiUon Oate: 

Original OalaPath: C:IXCALIBURIDATA\kaneko\018 SampleiD: 

6\FR 

Sample Name: Std. 0.0483 mgJL Comments: Sample rype: Std Bracket Operator. 

"' ANALYTICAL CONOtnONS (HPLC) "' 

HPLC TYPE : SHIMAOZU LC-20AD COLUMN : Hypetsl GOLD 2.1 >50 mm (Sum) 

ELUENT : A; 20mMAmmon!um fonnalcaq./form!cacld (1000:1) 

B; Acelonltrllo 

AIB(1:1) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Tharmo Asher Seienlilic TSQ. Quantum AcceS3 MAX 

IONIZATION : Negallve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : o CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 180) 

"''" 

"""' ''

38 
Recovery test 

(Fish analysis) 

Added Standard 

solution 

Recovery test 

(Fish analysis) 

Added group 

No.1 

Data File: C1S6FRAS Acquisition Dal&: Otlginal Data Path: C:\XCALIBUR\DATA\kanoko\C1S SamploiO: 

6\FR Samplo Name: Added Std. Comments: 

Samplv Typo: Unknown Operafor: 

"' ANALYTICALCONDITIONS(HPLC) '" 

HPLC 7YPE : SHJMADZU LC.ZOAD COLUMN HypetsU GOLD 2.1 x SO mm (5 urn) ELUENT A; 20 mM Ammonium formate aq./form!c acid (1000:1) 

B;Aeetonlkilo 

FLOWRATE : ~(~8ru, COLUMN TEMP : 313 K 

INJECT VOL : 6 uL 

'" ANALYTICAL CONDITIONS (MS) '" MS TYPE ! Thermo Fisher Sclenllfio TSQ Quantum Access MAX IONIZATION : NegaUva ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2SOOV SHEATH GAS : 50 

AUXGAS : 15 JON SWEEP GAS : 0 CAPiLLARY TEMP : 493 K 

SCAN 7YPE : SIM (mlz 180) 

i 

··-

Data FiltH C185FAA1·1 Acqulsltfon Date; 

Otlgtnat Data•Path: C:\XCALIBUR\DATA\I<aneko\C18 Sampte!D: 

6\FR Sample Name: Added No.1 Commanls: Sample Type: Unknown Operator. 

"' ANAL YT!CAL CONDITIONS (HPLC) "' 

HPLC 7YPE SHIMADZU LC·20AD COLUMN Hypersll GOLD 2.1 x 50 mm (5 um) ELUENT A; 20 mM Ammonium ~rmale aq.lform!c acid (1000:1) 

B;Acaton!trllc 

FLOWRATE : ~(~Dmln COLUMN TEMP ! 313 K INJECTVO!. : 5ul 

"' AIIALYTICAL CONDITIONS (MS) "' 

MS 7YPE : Thenno Fisher Scientific TSQ Quantum Acress MAX IONIZATION : Nega!lva ESI VAPORIZER TEMP ! 723 K SPRAYVOLTAGE : 2SOOV SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN 7YPE : SIM (mtz 180) 

39 
Recovery test 

(Fish analysis) 

Added group 

No.2 

Recovery test 

(Fish analysis) 

Added group 

No.3 

Data Fifo: C186FRA2 Acqulslflon Da!e: 

Original Data Path: C:\XCAUBUR\OATA\l<aneko\018 Sample ID: 6\FR 

Sample Name: Added No.2 Comments: 

Sample Type: Unkno\\'tl Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC 1YPE : SHIMAOZU LC.20AO 

COLUMN : Hypersl GOLD 2.1 x 50 1r.m (5 um) 

ELUENT : A: 20 mMAmlron!um formate aqJformlc acld (1000:1) B:AcetcnitrHa 

AIB(1:1) FlOW RATE : 0.2 mlJmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

.., ANALYTICAL CONOmONS (MS) '" 

MS lYPS. : Thermo Fisher Scle.nUffc TSQ Quantum Access MAX IONIZATION : Negailve ESI VAPORIZER TEMP : 723 K 

SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPilLARY TEMP : 493 K 

SCAN TYPE : SIM (mlz 160) 

..

u 

Data Fllo! C186FRA3 Acquisition Date: Original Data Path: C:\XCALIBUR\DATA\kaneko\C18 Sample ID: 6\FR 

Sample Name: Added No.3 Comments: 

Sample Type: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) ••• 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COlUMN TEMP INJECT VOL 

SHIMAOZU LC-20AO 

Hyporsi GOLD 2.1X 50 mm (5 um) 

A; 20 mM Ammonium formate aqJforrnlo acld (1000:1) 

B;Ao<!lonl'.r!le 

AIB(1:1) 

: 0.2mUm~ 

: 313K 

l Sul 

''' ANALYTICALCONOmONS (MS) "' MS TYPE : Therrr.o Asher Scientific TSQ Quantum Access MAX IONIZATION l Negallw ESI VAPORIZER TEMP : 723 K 

SPRAYVOLTAGE l 2500V 

SHEATH GAS : 50 AUXGAS : i5 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 1 80) 

40 
Recovery test 

(Fish analysis) 

Control group 

No.1 

Recovery test 

(Fish analysis) 

Control group 

No.2 

DataFite: C165FRCH Acquls!Uon Date: 

Original Data Path: C:IXCALIBUR\DATA\I<aneko\C16 SamploiD: 

6\FR 

SampJa Nama: Cfrl No. f Comments: 

Sample Type: Unknovm Operator: 

"' ANALYTICALCONDITIONS(IiPLC) ·~ 

HPLCTYPE : SHIMADZU LC·20AO COLUMi'l 

ELUENT : Hypersll GOLD 2.1 x 50 mm (5 um) : A; 20 mM Ammonium formate aq./fonnlo ecld (1000:1) B:Acelonlttilc 

FLOW RATE COLUMN iEMP INJECT VOL 

NB(1:1) : 0.2mUmln : 3t3K : 5uL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : ThermoflsherScterrtir~eTSQQuao!umAccess MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 250GV SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : ~93 K SCAN TYPE : SIM (m/z160) 

DatafUe: C166FRC2 Acqulstuon Dale: 

Original Data Path: C:IXCALtBURIDATAikaneko\C16 SampleiO: 

6\FR 

Sample Name: Ctrf No.2 Comments: 

Samplo. Type: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) m 

HPLCTYPE SHIMA02U LC.20AD COLUMN 

ELUENT Hyperolf GOLD 2.1 x 50 mm (5 um) A; 20 mM Ammonium formate aqlform!o acid (1000:1) a: Acetonffrlfa 

fl.OWRATE 

COLUMN TEMP 

INJECT VOL 

NB(1:1) : 0.2mUmin : 313K : 5 uL 

H' ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Fisher Sci.!lnL'fic TSQ Quanlum Access MAX IONIZATION : Negative ESJ VAPORIZERTEMP: 723K SPRAY VOLTAGE : 2500 V 

SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARYiEMP : 493K SCAN TYPE : SIM (rr!z 180) 

41 
Recovery test 

(Fish analysis) 

Control group 

No.3 

Dataflre: CI86FRC3 Acquisfl!on Date: 

Original Data Path: ~CAUBURIDATA1l<aneko\C18 SampieJD: 

SampJe Name: C!rl No.3 Comments: 

Sample Typo: Unknown Operator: 

......... : ....................... ._ ....................................................... .. '" ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE : SHIMAOZU LC-21lAD COlUMN : HypersiiGOLO 2.1xo0mm(5um) ELIIENT : A;20mMAmrnon!umformale"'lJformicaeld(1000:!) 

B;Aceton!trlla 

FLOW RATE : 'ri:~(~Bmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICALCONOtnONS (MS)"' 

MS TYPE : Thermo Fisher Sclenfir!C TSQ Quanfiim Access MAX IONIZATION ! Nega!lve ESI 

VAPORIZERTEMP: 723K SPRAY VOLTAGE : 2500 V 

SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP ! 493 K SCAN TYPE : SIM (mlz 180) 

'""'· tl

42 
7-2 Chromatograms of for recovery rate from water 

Recovery test 

(Water analysis) 

Standard solution 

0.0048 mgL-1 

Recovery test 

(Water analysis) 

Standard solution 

0.0483 mgL-1 

Oata fife: C166WRHS1·2 Acqu1sfUon Data: Original Dala Palh: ~jfca!lbu!ldala\kaneko\C1B6\W SamploiD: 

Sample Name: Std. 0.0048 mgfL Commen!s: 

Sampfa Type: Sfd Brac!ret Operator. 

........................................................................................... "' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC-20AD 

COLUMN : HypersU GOLD 2.1 X 50 mm (5 um) 

ELUENT ~i~~~~~moolum formate sqJiorm!c acld (1000:1) 

Nll(1:1) FlOW RATE : 0.2 mUm!n COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thsrmo Fisher SclentifJC TSQ Quantum Access MAX IONIZATION : NegaUve ESI VAPORIZER TEMP : 7231< SPRAYVOLTAGE : 2500V SHEATH GAS : EO AUXGAS : 15 ION SWEEP GAS : 0 CAPILlARY TEMP : 493 K 

SCAN TYPE : SIM (m'z 180) 

''''''"''"''''''''''"''"'''''''"'"''''''''""''''''"''''"'''''"''"'''''"''''"Hooouooohoo"'"''"'''"'''"'''''''' 

OafaFl1e: C186WRHS2·2 Acqubltro11 Date: 

Original Da!a Path: C:\Xcafibur\data\kaneko\C16tAW Sample ID: 

RH Sample Name: Std. 0.0483 mgit Commenls: 

Sample Type: S1d Btacket Opcrnlor: 

"'ANALYTICAL CONDITIONS (HPLC) 'M 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMADZU LC-20AD 

: Hypersa GOLD 2.1 x 50 mm (5 um) : A: 20 mM Ammonium formalo aq.lformlo acid (1000:1) B: Aoelonllrlle 

FlOW RATE 

COLUMN TEMP INJECT VOL 

Nll(1:1) 

: 0.2mUm!n : S1SK : 5UL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Asher Scientific TSQ Q'Jantum Access MAX 

IONIZATION : Negative ESI 

VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V 

SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m'z 180) 

RT:.C.Oih:1b2 ut1$0 

"""' ,.,.,. 

43 
Recovery test 

(Water analysis) 

High exposure level 

Added Standard 

solution 

Recovery test 

(Water analysis) 

High exposure level 

Added group No.1 

Data Fila: Original Data Path: 

Sample Name: Sample Type: 

C186WRHAS·2 

C:\Xcallburldalalkaneko\C 186\W 

RH 

Added Sid. 

Unknown 

"' ANALYTICAL CONDITIONS (HPLC) "' 

Acqufsltlon Oafe: 

SampleiO: 

Comments: 

Operator: 

HPLCTYPe 

COLUMN 

ELUENT 

SHIMAOZU LC.20AO Hyper>UGOLO 2.1 x60mm(5um) 

A; 20 mM Ammonium fo1111ale aqlfOflllllacld (1000:1) 

B: Acalon!We 

FLOW RATE COLUMN TEMP INJEOTVOL 

A/8(1:1) 

; 0.2mUmln : 313K 

: 5ul 

'" ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher SclenUfio TSQ Quantum Access MAX 

IONIZATION : NegaUva ESI VAPORIZER TE\lP : 723 K SPRAYVOLTAGE : 250DV SHEATHGAS ; 50 

AUXGAS : 15 

ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

u

Dala Fila: C186WRHA1·2 Aequts!Uon Date: 

Orlglnal Data Path: C:\Xl:allburldala\kaneko\C1BS\W SamploiD: RH Sample Name: Added No.1 Comments: 

Sample Typo: Unknown Operator: 

............................................................................................ '" Af/AL YTICAL COf/DITIONS (HPLC) "' 

HPLC TYPE : SHIMAOZU LC.20AD 

COLUMN : h)persHGOLO 2.1x50mm(5um) 

ELUENT : A; 20 mM Ammonium fo1mate aqJformlc acld (1000:1) B: Acetonitrile NB(1:1) FLOWRATE : 0.2mllmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'"ANALYTICAL CONDITIONS {MS) "' 

MS 1YPE : Thermo Fishe-r Sdtmtll1c TSQ Ol.mnhlm Acce.ss ,.,tAX IONIZATION : NegaUve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 

ION SWEEP GAS : 0 CAFILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

.................................................................................................................................... 

"""' 

44 
Recovery test 

(Water analysis) 

High exposure level 

Added group No.2 

Recovery test 

(Water analysis) 

High exposure level 

Added group No.3 

Oafa Frte: 

Original Oala Path: C1SSWRHA2·2 Aequt.tUon Date< 

~lfcaliburldala\l<aneko\C186\W Sample 10: 

Sample Name: Added No.2 Comments: Sample Type: Unknown Opa.ralon 

"' ANALYTICAL C0,1DITJONS (HPLC) "' 

HPLC TYPE : SHIMADZU Lc-20AO COLUMN : J{yparsl GOLD 2.1 x 50 mm (5 urn) ELUENT : A;20mMAmmenlumformaleaqJfotmloacld(100D:1) 

a;Acelonllrila 

!VIl(1:1) FLOW RATE : 0.2 mllmln COLUMN TSMP : 313 I< INJECTVOL : 5uL 

·~ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Fisher Sclentiflc TSQ Quanlum Access MAX IONIZATION : Negaiivo ESI VAPORIZER TEMP : 7231( SPAAYVOLTAGE : 2500V SHEATHGAS : 5<l AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (miz 1 eo) 

Data Fife: Original Data Path! C186WRHA3·2 

CX<cali1lu!lda!alkanel<o\C1Bfii'l/ 

RH Sample Name: SamploTypo: Added No.3 

Unknown 

'" ANALYTICAL CONDITIONS (HPLC) "' 

: SHIMADZU LC.20AO 

: J{ypmUGOLO 2.h50mml5um) 

Acquisition Oato: 

SamplalO: 

Comments: 

Operator: 

HPLCTYPE 

COLUMN 

ELUENT : A; 20 mM Ammonium founate aqJfotmlo acid (1000:1) B: AcelonlkHa 

FLOWAATE 

COLUMN TEMP INJECT VOL 

!VIl(1:1) : 0.2mUmln : 313 K : 5uL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclenfiria TSQ Quanlum Act:es' MAX IONIZATION : Negallve ESI VAPORIZER TEMP : 723 K SPAAYVOLTAGE : 2500V 

SHEATH GAS : 50 AUXGAS : 15 IONSWEEPGAS : 0 CAPILI.ARY TEMP : 493 K SCAN TYPE : SIM (mh ISO) 

45 
Recovery test 

(Water analysis) 

High exposure level 

Control group No.I 

Recovery test 

(Water analysis) 

High exposure level 

Control group No.2 

O.afaFito: C166WRHC1-2 Acquisition Date: Original Dala Palh: C:\XIl<lubui\dala\l<aneko\C166\W SamplaiD: RH Sampla Name: Clrl No.1 Commants: Sample Type: Unknown Operator: 

•~ ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMAOZU LC20AD COLUMN : Hyper.UGOLD 2.1 x50mm(5um) 

ELUENT : A; 20 mM Ammonium formate aq./fc!mtc acid (1000:1) 

B;Acelonllrlle 

N8(1:1) PLOW RATE : 0.2 m!Jmin COLUMN TEMP ; 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDITIONS (MS) "' MS lYPE : Thfrmo Fisher SctenUf!C TSQ Quantum Access MAX IONIZATION : Negative ESI 

VAPORIZER TEMP : 723 K SPRAYVDLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (11'Jz 150) 

U,

Data File: C1B6WRHC2·2 Acquisition Date: Orlgtnal Data Path: C:\Xcalibur\data\kaneko\C1B6\W Sample fO: RH Sample Name: Cfrl No.2 Comments: 

Sample Type: Unknown Operator: 

'"ANALYTICAL CONDITIONS (HPLC) '" 

HPLCTYPE 

COLUMN 

ELUENT 

SHIMAOZU LC20AD 

: HypersiiGOLD 2.1x50mm(5um) 

: A;20 mM Ammonium formate aqJ!ormlc acid (1000:1) 

a; Aceton!!rile 

PLOW RATE COLUMN TEMP INJECT VOL 

AIB(1:1) : 0.2rnUmln : 313K :5uL 

"'ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher SO.nUfio TSQ Quanlcm A~ss MAX IONIZATION : Negative ESI VAPORIZER TEMP ; n3 K SPRAY VOLTAGE ; 2500 V SHEATHGAS ; BO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 1 BO) 

ttr:o.n.::r.w

-,.,.,. 

"'''"

46 
Recovery test 

(Water analysis) 

High exposure level 

Control group No.3 

Recovery test 

(Water analysis) 

Standard solution 

0.0048 mgL"1 

Data File: C186WRHC3 .. 2 AcquisfUon Date: Original Oala Path: C:\XcaibUI\dala\kaneko\C186\W Sampla!O: RH Sample Nama: Clrt No.3 Comments: 

Sample Type: Unknown Operator: 

'" ANAL YT!CAL CONDITIONS (HPLC) '" 

HPLC TYPE SH!MADZU LG-20AD 

COLUMN HypersU GOLD 2.1 X50mm (5um) ELUENT A; 20 mMAmmon!um lonnaleaqJfOill11c ecld (1000:1) 

SjAI;elonilrifa 

FLOW RATE COLUMN TEMP INJECT VOL 

NB{1:1) : 0.2mUmin : 313K 

: SoL 

"' AIIALYT!CALCOND!T!ONS(MS) "' MS TYPE : Thermo Asher Sclenfifrc TSQ Quanlum Access t.iAX IONIZATION : Negative ES! VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPilLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

l'tT;o;tt.:uo 

Data File: C186WRLS1·2 Acqufsltlon Date; 

Original Data Path: C:\Xca!lbur\dala\kanako\C186\W SamplelD: RL Sample Name! Std. 0.0048 mgll Comments: 

Sampla Type: Std Bracket Oporalor: 

"' ANALYTICAlCOND!TlONS(HPLC) '" 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMADZU LC.20AD : HypersH GOLD 2.1 X 50 mm (5 um) : A; 20 mM Ammonium formale aqlformf<: acid (1000:1) B; AcetonJ!rflo 

FLOW RATE COLUMN TEMP 

INJECT VOL 

A/8(1:1) 

: 0.2mUmln : 313K 

: 5uL 

'" ANALYTICAL COIIDlT!ONS (MS) "* MS TYPE : Thermo Fisher S"Clanl!fic TSQ Ouanlum Atcess MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 60 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SlM (mlz 180) 

................................................................................................................................... 

47 
Recovery test 

(Water analysis) 

Standard solution 

0.0483 mgL"1 

Recovery test 

(Water analysis) 

Low exposure level 

Added Standard 

solution 

Dala File: C166WRLS2·2 Acquisition Date: Original Data Path: C:\Xcalibur'dala\kaneko\Ctll6\W Sample 10: RL Sampf.a Name: Std. 0.0483 mglt Comments: 

Sample Type: Sid Bracket Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE : SHIMAOZU LC.20AO COLUMN 

ELUENf : Hypersll GOLD 2.1 xso mm (Sum) : A; 20 mM Ammonium form ale aqJiormlc acid (1000:1) 

B; Ace1oni!rlre 

FLOW RATE COLUMN TEMP INJECT VOL 

NB(1:1) 

; 0.2m\Jmln : 313K : 5 oL 

'" ANALYTICAL CONDITIONS (MS) '" MS 1YPE : Thermo Flsher SclenUlic TSQ Quantum Acteu MAX IONIZATION : Nejjalive ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : Sf~ (miz 1 BO) 

Data File: C166WRLA8-3 Acquls!Uon Dale: Original Data Path: C:\Xcallbur\da!alkaneko\C165\W Sample ID: RL Sample Name: Added Std. Comments: 

Sample Type: Unkrnwm Operator: 

"'ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE SHIMADZU LC.20AO COLUMN H)llersi GOLD 2.1 x SO mm (5 um} ElUENT A: 20 mM Ammonium formale aq./fctmlo acid (1000:1) 

B; Acetonltrifs NB(1:1) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 oL 

"" ANALYT!CALCONOITIONS(MS) "" MS TYPE : Therrr.o Fisher Sclentmc TSQ Quantum Access MAX IONIZATION : Negalive ESI VAPORIZER TEMP: 723K SFRAYVOLTAGE : 2500V SHEATH GAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCANTYPE : SIM(rr/z1BO) 

48 
Recovery test 

(Water analysis) 

Low exposure level 

Added group No.1 

Recovery test 

(Water analysis) 

Low exposure level 

Added group No.2 

Data File: C166WRLA1-4 AcqulsiUon Date: 

Original Data Path: C:\Xcallbul\data\IQlneko\C1BS\W Sample!O: 

Rl Sample Namo: Added No.1 Comments: 

Sampta Typa: Un!mov.n Operator: 

'" ANALYTICAL CONDITIONS (HPLC) bo 

HPLC lYPE : SHIMADZU LC·20AO COLUMN : Hypersll GOLD 2.1 x 5ll mm (5 om) ELUENT : A; 20 mMAmmon~.1111 furmale aq./formlc ocld (1000:1) Bi Acetonitrile AIB(1:1) FLOWRATE : 0.2mUmfn 

COLUMN TEMP : 313K INJECTVOL : 5Ul 

"' ANALYTtCALCONOITIONS(MS) "' MS TYf'E : Thermo Flsher Sctenlif1c TSQ Quantum Access A-fAX IONIZATION ; Negalive ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE ; SIM (mk 160) 

-

... 

Oata Flle: C186WRLA2~2 Acquisition Date: 

Original Data Path: C:\Xcalibul\dalalkaneko\C1B6\W Sample 10: RL Sampfe Name: Added No.2 Comments: 

Sample Typo: Unknown Operator: 

"' ANAL YTlCAL CONDITIONS (HPLC) "' 

Hf'LC TYPE : SHIMADZU LC-2MD COLUMN : Hypersi GOLD 2.1 x 50 mm (5 um) 

ELUENT : A; 20 mM Ammonium formate aqJformlc acid (1000:1) 

e; AcelonllrUa AIB(1;1) 

FLOW RATE : 0.2 mllmln COLUMN TEMP : 313 K tNJECTVOL : 5Ul 

"' ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : TlMlrmo Asher Sclent!trc TSQ Quanlt.lm Access MAX IONIZATION : NegaUva ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 160) 

49 
Recovery test 

(Water analysis) 

Low exposure level 

Added group No.3 

Recovery test 

(Water analysis) 

Low exposure level 

Control group No.1 

Data File: C186WRlA3·2 Acqulslllon Date: 

Original Data Path: C:\Xcaiburldala\kaneko\C186\W Samp!o ID: 

RL Samplo Uame: Added No.3 Comments: 

Sample Type: Unknown Operator: 

'" ANALYTICAL CONDITIONS (IIPLC) "' 

HPLC TYPE SHIMAOW lC-20AD COLUMN Hyp""'U GOlD 2.1 X 50 mm (5 urn) ELUENT A: 20 mM Ammonium formal<! aqlformlo acid (1000:1) 

Bi Acetonitrile AIB(1:1) 

FLOW RATE : 0.2 mllmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

•~* ANALYTICAL CONOrrtONS {MS) •u 

MS TYPE : Thermo Fisher Sclentinc TSQ Quantum Access MAX IONIZATION : Negallve ESI VAPORIZER TEMP: 723K SPRAYVOLTAGE : 2SOOV SHEATH GAS : 50· AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 180) 

Data File: C186WRLC1-3 Acquisition Date: 

Original Data Path: C:\Xca!!burldnla\l<aneko\C186\W Sample ID: 

Rl Sample Nama: Cfrl No.1 Commonfs: Sample Typo: Unknown operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

H?LC TYPE : SHIMAOZV lC·20AD 

COLUMN : Hyporsll GOlD 2.1 X 50 mm (5 um) 

ELUENT : A;20 mMAmmonlum formate aqJfotrnlc acid (1000:1) B;Aceton!tnle 

AIB(1:1) FLOWRATE : 0.2ml!mln COLUMN TEMP : 313K !NJECTVOL : 5uL 

"' ANALYTICAL CONDITIONS (MS) '" MS TYPE : Thermo Fisher Sclenllnc TSQ Qua:nlum Access MAX IONIZATION : !legallvo ESI VAPORIZER TEMP : 723 K 

SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPillARY TEMP : 493 K 

SCANTYPE : SIM(mlz180) 

RT:O.~·:tco &.t15G 

'"'"" 

50 
Recovery test 

(Water analysis) 

Low exposure level 

Control group No.2 

Recovery test 

(Water analysis) 

Low exposure level 

Control group No.3 

Data FUe: C1B6WRLC2·2 Acquisition Dale: 

Original Data Path: C:\Xcalibui\da!all<aneko\C186\W SamploiO: RL Sample Name: Ctrl No.2 Comments: 

Sample Type: Unknown Operator: 

'" ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE : SHIMADZU LC·20AD COLUMN : Hyp<lrsU GOLD 2.1 X 50 mm (5 um) ELUENT : A;20mMAmmonlumfonnaleaqlfOlmlcacld(1000:1) 

B: AcelonittUe AIB(1:1) 

FLO\VRATE : 0.2mtlmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Ftsher Sclentlfic TSQ Quantum Access MAX IONIZATION : Negative ESI VAFORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAFILLAAYTEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Data FUe: C186WRLC3--2 Acquisition Date: Orlgfnal Data Path: ~L\Xcaflbu!\dala\kaneko\C1S6\W Sample 10; 

Sampla Namo: Ctr! No.3 Comments: Sampfo Typo: Unknown Operator: 

............................................................................................. , 

"' ANALYTICAL CONDITIONS (HPLC) '" 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMAOZU LC.20AD : Hypersll GCLO 2.1 X 50 mm (5 um) 

: A; 20 mM Ammcn!um fOrmate aq./fonnic ecld (1000:1) DiAtelon!lrilo 

FLOW RATE COLUMN TEMP INJECT VOL 

AIB(1:1) : 0.2m!Jmtn : 313K :SuL 

•~ ANALYTICAL CONDITIONS (MS) "* MS TYPE : Thermo Asher Scleotifio TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATHGAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLMY TEMP ! 493 K SCAN TYPE : S~.l (mlz 180) 

•1

51 
Reference 8 

8-1 

Chromatograms in fish 

Test group 

.---------------· 

Fish analysis 

Day7 

Standard solution 

0.0048 mgL" 1 

Da!a Fifo: 

Original Da!a Path: 

Sampht Name: 

sample Typo: 

C186WFA7S1 Acqu!siUon Dato: 

C:\Xcallbur\Oa!o\kaneko\C185\Da Sample 10: y7 

Sid. 0.0048 mgll Comment.: 

Std Brackal Operator: 

'" ANALYTICALCONDITIONS(HPLC} '" 

HPLC TYPE : SHIMADZU LC-20AD COLUMN : Hypersll GOlD 2.1 X 50 mm (5 urn} 

ElUENT : A; 20 mM Ammonlum formate aq.tronnlc acid (1000:1} B;Acelonftrife 

Alll(1:1) 

FLOW AATE : 0.2 mUrnln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

'" ANAl VTICAl CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Sciat.tifio TSQ Quantum Aa;ess MAX IONIZATION : Nega!lw ESJ VAPORIZER TEMP : 723 K SPAAVVOLTAGE : 25COV SHEATH GAS : 50 AUXGAS ; 15 ION SWEEP GAS : 0 CAPILLARYTEMP : 493 K SCAN TYPE : SIM (rnlz 180} 

Fish analysis 

Day7 

Standard solution 

0.0964 mgL·1 

Data Filo: C1B6WFA7S2. At"qutsltton Date: 

Orlgfnal Data Path: C:\XcaUbur\Data\kaneko\C186\0a Sample fO: y7 

Sample Name: ·std. 0.0964 mg/L Comments; 

Sample Typo! Std Bracket Operator: 

"'ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMAOZU lC-20AO COLUMN : Hypersll GOLD 2.1 x 50 mm (5 um) 

ELUEIIT : A; 20 mM Ammonium formate aq.trorrnlo acid (1000:1} B;Acelonl~llo 

NB(1:1} FLOW AATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOl : 5 ul 

'" ANAlYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclenlifit: TSQ Quanlum ACCe$S MAX IONIZATION : NsgaUvo ESI VAPORIZER TEMP : 723 K SPAAVVOLTAGE : 2500V SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPillARY TEMP : 493 K SCAN TYPE : SIM (rnlz 180} 

52 
Fish analysis 

Day7 

High exposure level 

No.1 

Fish analysis 

Day7 

High exposure level 

No.2 

Dafaflle: C1B6FA7H1 Acquisition Date: 

Original Data Path: C;\Xtallbur\Data\l<aneko\CIB6\0a Sample ID: y7 Sample Namo: Ash anelysls H lv No.1 Commanfs: Sample Type: Unknown Op~rator: 

"' ANALYTICAL CONDITIONS (HPlC) '" 

HPlCTYPE : SHIMADZU lC-20AO COlUMN 

ElUENT : Hyporsll GOlD 2.1 x 50 mm (5 urn) 

A; 20 mM Ammonium formale aqJformlo acid (1000:1) 

B;Acelonltraa 

FlOW RATE COlUMi'llEMP 

INJECT VOl 

Ai!l(1:1) : 0.2mlfmln 

: 313K : 5uL 

"' ANAlYTICAL CONDITIONS (MS) '" MS TYPE : Thermo Fisher Sclenllfic TSQ Quantum Access MAX IONIZATION : Negallva ESI VAPORIZERTEMP: 723K 

SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Data File: C18SFA7H2 Acquls!Uon Dale: Original Oat a Path: C:\Xca11bur\Data\kaneko\C1 66\0a Samplo.IO: 

y7 

Sampto Name: Fish analysls·H lv·No.2 Comments: Sample Type: Unknown Oparator. 

"' ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE : SHIMADZU lC-20AD COlUMN : Hypersll GOLD 2.1 x SO mm (5 um) ELUENT : A; 20 mMAmmon!um fonnale aqJfoll!11c acid (1000:1) a: Acelonitn1e NS(1:1) FlOWRATE : 0.2mUmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"' ANALYTICAL CONDITIONS (MS) -

MS TYPE : Thermo Fisher Scientific TSQ Quanlum Access MAX IONIZATION : NegaUve ESI VAPORIZER TEMP : 723 K SPRAYVOlTAGE : 2SOOV SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 1 SO) 

RT:o.c¢·~v.t:1SG 

lCCIOOOO 

'"'''" 

53 
Fish analysis 

Day? 

Low exposure level 

No. I 

Fish analysis 

Day? 

Low exposure level 

No.2 

Oala File: C1S6FA7L1 Acqulsl!lon Date: 

Orlglnal Data Path: C:\XcaHbu~Oata\l<aneko\C1S6\Da SamploiD: 

y7 

Sample Name: FJsh analysis L lv No.1 Comments~ 

Sampte Typo: Unknown Opuator: 

"' ANALYTICAL CONDITIONS {HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMADZU LC-20AD : Hypersll GOLD 2.1 x 50 mm (5 urn) 

A; 20 mMAmmonlum formate aqJformlo acld (1000:1) S;Acelonllille 

FLOW RATE COLUMN TEMP 

INJECT VOL 

AIB(1:1) : 0.2mUmin : 313K 

: 5UL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Asher Sclcntlria TSQ Quantum Access MAX IONIZATION : NegaUvo ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2oOOV SHEATHGAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCANTYPE : SIM(mlz1SO) 

1\TIO.tc~).OO SltUG 

"""' '"'""' 

ur

Data File: C186FA7l2 Acquisition Dato: 

OrlgtnaJ Data Path: 

Sampla Nama: 

C:\Xcafl:bur\Oala\kaneko\C186\0a Sample fD: 

y7 

Fish analysis l Lv No.2 Comments: 

Sample Type: Unknown Operator; 

"' ANALYTJCALCONDJTIONS(HPLC) '" 

HPLC TYPE : SHIMADZU Lc-20AD COLUMN : HypernO GOLD 2.1 X 50 mm (5 um) ELUENT : A; 20 mM Ammonium format a aq.lformlc acid (1C00:1) 

FLOW RATE 

COLUMN TEMP INJECT VOl 

B; AcetonllrHa NB(1:1) 

: 0.2mUmln : 313K ! 5UL 

"' AIIAL YTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Sc{entific TSQ Quan!um Access MAX IONIZATION : Naga!lve ESI 

VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

------

54 
Fish analysis 

Day 14 

Standard solution 

0.0048 rngL·' 

Fish analysis 

Day 14 

Standard solution 

0.0964 rngL·' 

Data FOe: C186WFA14S1 Acquisition Date: Original Data Path: C:\Xcallbut\Da!a\keneko\C186\Da Samp!eiO: y14 Sample Name: Std. 0.0048 mg/L Comments: 

Sample Typa: Sid Erackat Operalor: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMADZU lC.20AO : Hyporsi GOlD 2.1 x 50 mm (5 urn) : A; 20 mM Ammonium formate aqJformlo add (1000:1) 

B; Acetonlll!le Al!l(1:1) : 0.2mUmln : 313K : 5Ul 

'" ANALYTICAL CONDil!ONS (MS) m 

MS TYPE : Thermo FlsherSc!entlllcTSQ Quantum Access MAX IONIZATION : Negal!va ESt VAPORIZER TEMP : 723 K SPAAYVOLTAGE : 2SOOV SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m'z 180) 

Rf:O.C¢-:).00 .SIA:U3 

u~

~c.o'

Data File: C1S6WFA14S2 Acquisition Date: Original Data Path: C:\Xcalibut\Data\kanelro\C186\Da SampleiD: 

y14 

Sample Name: Std. O.OSt64 mg!L Comments: 

Sample Type: Std Brac.~et Operator. 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

SHIMADZU LC.20AD Hypersll GOLD 2.1 x 50 mm (5 um) A; 20 mMAmmonlum fonnale oqJfonnlc acid (1000:1) Bi Acetonlln1e 

:-:<~]min : 313K 

: 5ul 

'" ANAL YTlCAL CONDITIONS (MS) •ri 

fi,1S TYPE : Thermo Fisher Scfenlific TSQ Quantum Access MAX IONIZATION : Negallve ESI VAPCRIZERTEMP: 723K SPAAYVOLTAGE : 2500V SHEATH GAS : 60 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

1\1; 0.00~),00 $l.t t$!J 

""" """ 

~~

55 
Fish analysis 

Day 14 

High exposure level 

No. I 

Fish analysis 

Day 14 

High exposure level 

No.2 

Data File: C186FA14H1 Acqulslllcn Date: 

Original Data Path: C:\Xcallbu!l!lata\kaneko\C186\0a SamploiO: 

y14 

Sampta Name: Fish analysis H Lv No.1 Comments: 

Sample Type: UnJr.nown Operator: 

HO ANALYTICAL CONDITIONS (HPLC) ... 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

SHIMAOZU LC.20AO Hypersil GOLD 2.1 X 50 mm (5 urn) 

A; 20 mM Ammonium formate aq.Jformlc acid (1000:1) B;AC1llonltrllo 

NB(1:1) : 0.2mUmln : 313K : 5ul 

•~ ANALYTICALCONDITIONS(MS) '" 

MS TYPE : Thenno Fisher Sclenllilc TSQ Quantum Access MAX IONIZATION : NegaUvo ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

U

Oatafllo: C185FA14H2 Acqulsl!lonOato: 

Original Data Path: C:\Xcalibw\OatsUianeko\C186\Da Sample ID: y14 

Sample Nam-e:: Fish analysts H lv N-o.2 Comments: Sample Type: Unknown Operator: 

"' ANALYTICAL COflDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMAOZIJ LC·20AD HypersU GOLD 2.1 x 50 mm (5 um) A; 20 mMAmmonlum fotmale eq.Ifotmlc ecld (1000:1) 

B; AcelonUrile NB(1:1) : 0.2mUmin : 3i3K : 5ul 

"' ANALYTICAL CONDITIONS (MS) '" MS 1YPE : Thermo Fisher Sclenllfic TSQ Quantum Access MAX 

IONIZATION : NegoUve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : EO AUXGAS : 15 ION SWEEP GA9 : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

o

56 
Fish analysis 

Day 14 

Low exposure level 

No. I 

Fish analysis 

Day 14 

Low exposure level 

No.2 

Data File: 

Original Dafa Path: 

Sampla Nama: Sample Type: 

C186FA 14L 1 Acqulsll!on Date: C:\Xcalibur\Dala\kaneko\C1S6\Da Sample ID: yH 

Fish analysts L lv No.1 Commenfs: Unknown Operator. 

'" ANALYTICALCOND!TIONS(HPLC) "' 

HPLC TYPE : SHIMAOZU LC·2DAO 

COLUMN : Hyparsil GOlD 2.1 x 50 mm (5 urn) ELUENT : A; 20 mM Ammonium formate aqJTormlo acid (1 000:1) 8; Acefonitrlia AIB(1:1) FLOW RATE : 0.2 mUmln COlUMN TEMP : 313 K INJECT VOL ; 5 uL 

'" ANALYTICAL COilDITIONS (MS) "' 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Naga!lva ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500 V SHEATHGAS ; 60 

AUXGAS : 15 ION SWEEP GAS : 0 

CAPillARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

............................. u ................................................................................................... . 

Data File: C186FA14l2 Acquisition Date: 

Orlglnal Dala Path: 

Sample Name: 

C:\XcaU!rur\Data\kanako\C186\Da SamplaiD: 

y14 

Frsh analysis L Lv No.2 Comments: Sample Typo: Unknown Operator: 

"' ANALYTICAL CONDITIONS {HPLC) '" 

HPLCTYPE : SHII.'.ADZU LC.20AD COLUMN 

ElUENT ; Hypersll GOLD 2.1 x 50 mm (5 um) : A; 20 mM Ammonium forma to aqJformic ac!d (1000:1) 

B; Acetonlfrlla 

FLOW RATE COLUMN TEMP INJECT VOl 

A/8(1:1) : 0.2mUmin : 313 K : 5uL 

'" ANALYTICALCONDITIONS(MS) •u 

MS lYPE : Thermo Fisher Sci&ntific TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP: 723K SPRAYVOI.TAGE : 2500V SHEATH OAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mh: 180) 

57 
Fish analysis 

Day21 

Standard solution 

0.0048 mgL"1 

Fish analysis 

Day21 

Standard solution 

0.0964 mgL· 1 

Data Fllo: C166WFA2tS1 Aequfsltlon Dale: 

Original Data Palh: C:\Xcalibur\Oalalkaneko\0166\Da Sample ID: y21 Sample Nama! 

Sample Typo: Std. 0.0048 mg/L Comments: Std Broc-ket Operator: 

••• AMALYTICALCONOITIONS(HPLC) '" 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMADZU LC·20AD liyper;UGOLO 2.1 x50mm(5um) 

A; 20 mMAmmonlum formals aqJformlc acid (1000:1) 

B;Acelonlln1e 

FLOW RATE COLUMN TEMP INJECT VOL 

NB(1:1) 

: 0.2mUmlo : 313K :5Ul 

'" ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Fisher S<:!en~fio TSQ Quantum Aceess MAX IONIZATION : N'!)allvo ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARYTEMP : 493 K SCAN TYPE : SIM (m/z 180) 

Data Fife: 

Original Dala Path: 

Sample Name: 

Sample Type: 

C186WFA21S2 AequlsiUon Dale: C:\XcallburlDala\kaneko\C18S\Da Srunp!o!D: 

y21 

Sid. 0.0964 mg/L Comments: 

Sid Bracket Operator: 

wo ANALYTICAL CONDITIONS {HPLC) '" 

HPLC TYPE SHIMADZU LC·20AO COLUMN liypersH GOLD 2.1 x 50 mm (5 um) 

ELUENT A: 20 mM Ammonium fonnale eqJformlo acid (1000:1) 

B; Acotonllrllo A/6(1:1) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

••• ANALYl1CAL CONDITIONS (11.5) •ri 

MS TYPE : Thenno Fisher Scientific TSQ Quantum Access Mt-.X IONIZATION : NegaUva ESI VAPORIZER TEMP ; 723 K SPRAYVOLTAGE : 2500V 

SHEATHGAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPIULARY TEMP : 493 K SCAN TYPE : SIM (mlz ISO) 

~.l

58 
Fish analysis 

Day21 

High exposure level 

No.! 

Fish analysis 

Day21 

High exposure level 

No.2 

Data File: C186FA21H1 Acqulsltfon Dale: Original Oala Path: C:\XcallbU!IDala\kaneko\C186\0a SampleiD: 

y21 Sampfo Name! Ffsh ana.fysfs H Lv No.1 Comments: 

Sampfo Typo: Unknown Oporafor: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLC1YPE : SHIMADZU LC.20AO COLUMN 

ElUENT Hypersl GCLO 2.1 x 60 mm (5 um) A; 20 mM Ammonium formate aqJformlc acid (1000:1) Bi Acelonifrlle 

FLOW RATE COLUMN TEMP INJECT VOL 

NB(1:1) : 0.2mUmin : 313K : 5ul 

H. ANALYTICAL CONDITIONS (MS) •» MS lYPE : Thermo Fisher Scientific TSQ QUanlum Access MAX 

IONlZATlON : Negallve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V 

SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mil 160) 

nr:o.oo.uo 

'"""' ,...,. 

___

Data File: C185FA21H2 AcqutslUon Date: 

Original Data Path: C!\Xca4'but\Oata\kaneko\C186\0a Sampte 10: 

y21 Sample Na:me: Fish analysis: H lv No.2 Comments: Sample Typo: Uni'.nown Operator: 

'" ANAL YT!CAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMADZU LC.20AD : Hype!Sll GOLD 2.1 x 50 mm (5 um) : Ai 20 mMAmmonlum fc:mc:te aqJformic ae-!d (1000:1} B; Acelonltrile 

FLO\'/ RATE 

COLUMN TEMP INJECT VOL 

Ml(1:1) 

: 0.2mUmln : 313K : 5UL 

m AIIALYTICALCONDITIONS (MS) "' 

~.iS TYPE : Thermo Ffsher Sclentllio 1SQ Quanlum Access MAX lONlZATION : NegaUve ESI VAPORIZER TEMP: 723K SPRAYVOLTAG;; : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARVTEMP : 493K SCAN TYPE : SIM (mlz 180) 

59 
Fish analysis 

Day21 

Low exposure level 

No. I 

Fish analysis 

Day 21 

Low exposure level 

No.2 

DalaFJJo: C166FA21 L 1 Acquisition Date: 

Orfglnal Data Palh: C:\Xcallbur\Data\kaneko\C166\Da Sample to: 

Y21 Sample Name: Fish analysis llv No.1 Comments: Sample Typo: Uokna.vn Operator: 

"' ANALYTICAL CONDITIONS (HPI.C) "' 

HPLCTYPE : SHIMADZU LC·20AO COLUMN 

ELUENT 

Hyparsll GOLD 2.1 x 50 mm (5 um) 

k. 20 mMAmmonlum f0!1!1ale aqHormlc a~d (1000:1) B;Aceloni!rila 

FLOW RATE COLUMN TEMP 

INJECT VOL 

N8(1:1) : 0.2mUmln : 313K :Sui. 

"' ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher SclenUfro TSQ Quanlum Access MAX. ION12ATION : Negallve ESI VAPORIZER TEMP : 72.l K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Dalaflle: 

Or(gfnal Data Palh: C1B6FA21L2 AcqulsiUon Date: 

C:\Xcafibtir\Data\kaneko\C186\0a SamplsiD: y21 Sampfe Name: 

Sample Type: Fish anaiJ.'Sis llv No.2 Ccmmonls: · 

Unkr.own Operator: ........................................................................................... , 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC.20AD 

COLUMN : Hyper.;U GOLD 2.1 x 50 mm (5 um) 

ElUENT : k. 20 mM AmmoniL'fll formola aqJformlo acid (1000:1) a; Ac-etonllrRo 

FLOWRATE : :<~Vmln COLUMN TEMP : 313 K INJECT VCL : 5 ul 

'" ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo F1sher Selent'!ic TSQ Quantum Act:ess MAX IONI2ATION : NegaUve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILI.ARYTEMP : 4~3 K 

SCAN TYPE : SIM (mlz 180) 

................................................................................................................................... 

.

60 
Fish analysis 

Day23 

Standard solution 

0.0048 mgL'1 

Fish analysis 

Day23 

Standard solution 

0.0964 mgL·1 

Data File: C186WFA23S1 Acqutslllon Date: 

Original Data Path: C:\Xcalibur\Daia\kaneko\C186\Da SompleiD: 

y23 

Sampro Name: Std. 0,0048 mg/L Commonfs: 

Sample Type: Sid Bracket Operator: 

"'ANALYTICAL CONDITIONS (HPLC) •~ 

HPLClYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEW.P INJECT VOL 

: SHIMAOZU LC·20AD : HypersKGOLD 2.1x50mm(5um) : A;20 mM Ammonium formalo nqJ!o<mlc acid (1000:1) a: Acelcnllillo A/8(1:1) 

: 0.2mUmfn : 3131< : 6uL 

"' ANALYTICAL COtllliTIONS (MS) '" MS TYPE : Thermo Fisher Scfenllfic TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP! 723 K SPRAYVOLTAGE ! 2500V SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mh: 180) 

""':

Data Flle: C186WFA23S2 Acquisition Dale: 

Original Data Pa!li: C:IXoalibiR\Data\kaneko\C186\Da Samplo 10: y23 

Sample Nama: Std. 0.0954 mgn.. Comments: 

Sample 'fYpo: Sid Bracket Operotor: 

OM ANALYTICAL CONDITIONS (HPLC) ... 

HPLOTYPE 

COLUMN 

aUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMADZU LC·20AO : Hypersll GOLD 2.1 X 50 mm (5 urn) : A; 20 mM Ammonium formale aqJformlc ecld (1000:1) 

B;Acelcnllrile 

AIB(1:1) : 0.2mUmln : 3131< : 5ul 

w• ANALYTICAL CONDITIONS {1\'.S) "' 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negalive ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : tro AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM {mlz 180) 

: 

61 
Fish analysis 

Day23 

High exposure level 

No.1 

Fish analysis 

Day 23 

High exposure level 

No.2 

Data File: C186FA23H1-1 Acquisition Date: Original Data Path: C:\XcallburlDala\kaneko\C186\0a Sample ID: 

y23 Sample Name: Fish analysis H Lv No.1 Comments: Sample Type: Unkrw....n Operator: 

'" ANALYTICAL CONDITIONS (HPLC) '" 

SHIMAOZU LC-20AO : HyporsifGOLO 2.1x50mm(5um) HPLCTYPE 

COLUMN 

ELUENT : A; 20 mM Ammonium formala aqJfarmlc acid (1000:1) B: Acaloo!UIIa 

FLOW RATE COLUMN TEMP INJECT VOL 

NB(1:1) : 0.2rnllmln 

: 313K : 5uL 

"'ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Flsbe:r Scientific TSQ Quanlt!m Access MAX IONIZATION : Nogallvo ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 160) 

Data File: C1B6FA23H2-1 Aequlslllon Data: 

Original Data Path: C:\Xcallbur\Oafa\kaneko\C186\Da Samplo 10: y 23 

Sample Name: Fish analysis H Lv No.2 Comments: Sample Type: Unknown Op~ralor: 

'" ANAL YriCAl CONDITIONS (HPLC) "' 

H?LC TYPE SHII.'ADZU LC-20AD 

COLUMN : Hyporsll GOLD 2.1 x 50 mm (5 urn) ELUENT : A: 20 mMAmmonlum formale aq./formlc acid (1000:1) 

B; AcafonUrl!a NB(1:1) FLOIVRATE : 0.2mUmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"'ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher SclentifJC TSQ Quantum Access MAX IONIZATION : NogaUve ESI VAPORIZERTEM?: 723K SPRAYVOLTAGE : 2500V SHEATH GAB : 60 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 1 SO) 

62 

.., 
Fish analysis 

Day23 

Low exposure level 

No. I 

Fish analysis 

Day23 

Low exposure level 

No.2 

OalaFIIe: 

Original Data Palh: 

Sample Name: 

Samplo Type: 

C186FA23L1-1 

C:\Xcallbur1Dala\kaool<o\C186\Da 

t1~ analys~ L Lv No.1 

Unknov.n 

"' ANALYTICAL CONDITIONS (HPI.C) "' 

HPLC ll'Pg SHIMADZU LC·20AD 

Acquisition Dato: Samplo!D: 

Comments: 

Operator: 

COLUMN Hypersll GOLD 2.1 x 50 mm (5 urn) ELUENT A: 20 mMAmmonlum formate aqlfonnlo acid (1000:1) B: Acetonltrllo NB(1:1) FLOW RATE : 0.2 mllmln COLUMN TEMP : 313 K INJECT VOL : 5 u!. 

"' ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Asher SclenU!ic TSQ Quantum Access MAX IONIZATION : NegaUva ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mil 180) 

Data File: C186FA23l2-1 Acqulslllon Date: Original Data Path: C:\Xca!bur\Data\lcanel<o\C186\0a SamplaiD: 

y23 

Sampla Name: Ffsh analysis L lv No.2 Comments: 

Sample Type: Unknown Operator: 

"' ANALYTICAl CONDITIONS {HPLC) "' 

HPLC TYPE : SHIMADZU LC-20AD COLUMN HypersH GOLD 2.1 X 60 mm (5 um) ELUENT A; 20 mM Ammonium formalo aq.lfonnlo ac!d (1000:1) 

B; Acetoru1rile NB(1:1) FLOW RATE : 0.2 mt.lmln COLUMN TEMP : 313 K INJECT VOL : 5 L-1. 

••• AIIAI.YTICALCONOITIONS (MS) '" 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 60 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN ll'PE : SIM (mlz 1 60) 

}Fi

63 
Fish analysis 

Day28 

Standard solution 

0.0048 mgL"1 

Fish analysis 

Day28 

Standard solution 

0.0964 mgL·1 

Data Fife: C186WFA28S1 Acqulslflon Date: 

Original Data Path: C:\Xcalibur\Oala\kaneko\C16S\Da Sampla 10: y28 

Sampfe Name: Sfd. 0.0046 mglt.. Comments: 

Sampfa: Type: Sid Bracket Operator: 

H• ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU Lc-20AO COLUMN : Hyperd GOLO 2.1 X 50 mm (5 um) 

ELUENT : A; 20 mM Ammonium forma to aq.lforrnlc ac!d (1000:1) 

B; Ac.elonilri!a 

FtOWRATE : ~~(;:;/)min COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclenllfio TSQ QuanlumAccess MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K 

SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Data Fila: C186WFA28S2-2 Acquisition Date: Original Data Path: C:lXcallilw\Oata\kaneko\C185\0a SampleiD: 

y28 

SampJe Name: Std. 0.0964·mg!L Comments! 

Sampre Type! Std Braekef Operator: 

'" ANALYTICAL CONDITIONS (HPLC) ... 

HPLC TYPE SHIMAOZU LC.20AO COLUMN Hypersil GOLD 2.1 x 50 mm (5 umj 

ELUENT A;20 mM Ammonium formalo aqJformlo a~'d (1000:1) B: Acelonllri!e AIB(1:1) FLOW RATE : 0.2 mtlmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

... ANALYTICAL CONDITIONS (MS) H• MS TYPE : Thermo Fisher Scleni!Rc TSQ Quantum Access MAX ION12ATION : NegaUve ESI VAPORIZER TEMP: 723 K SPRAYVOLTAGE : 25COV SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 160) 

""" ,. 

..

64 
Fish analysis 

Day28 

High exposure level 

No.1 

Fish analysis 

Day28 

High exposure level 

No.2 

DalaFlle: CIB6FA26HI Acquisition Dale: Original Data Path: 

Samplo Nama; 

Sample Typo: 

C:\Xcallbut\Data\kaneko\0186\Da Sampla 10: 

y2B 

Fish analysJs H Lv No.1 Comments: Unknown Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC.20AO 

~~~~/: f ~fo~Gk.~o~~~;:r:.~e<!:;l,m!cac!d(1000:1) B;Acolonlfrua 

Al8(1:1} FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAl CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher ScfenUfic TSQ Quan!um Access MAX 

IONIZATION : Negallva ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 1 60) 

OalaF!Je: 

Orfglnal Data Path: CI86FA26H2 Acquls!Uon Date: C:\Xcalibur\Dallle<aneko',C196\Da SampleiD: y28 Sample Name: Fish analysis H Lv No.2 Comments: Samplo Type! Unknown Operalor: 

"' ANALYTICAL CONDiliONS (HPLCJ '" 

H?LC TYPE SHIMAOZIJ LC.20AD 

COLUMN Hype"D GOLD 2.1 x 50 mm (5 urn) ELUENT A; 20 mMAmmonlum formate aqJformlc acid (1000:1} B;AcalllnllrUa 

Alil(l:l) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313K INJECT VOL : 5 uL 

"" ANALYTICAlCONDmONS(MS} "' t.m TYPE : 1hflrrno Fisher Scle:nlific TSQ Quantum Access MAX IONIZATION : Negaliva ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATHGAS : 56 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K 

SCANTYPE : Sh\1(rnlz180) 

................................................................................................................................... RnC.t<l•'-t<llt1.tlSC 

'""" '""" 

0· 

a 

65 
Fish analysis 

Day28 

Low exposure level 

No. I 

Fish analysis 

Day28 

Low exposure level 

No.2 

Data File: C1B6FA2BL 1 Acquisition Date: 

Original Data Path: 

Sample Name: 

C:\Xcallbur\Oala\kaneko\C186\Da SampleiD: 

y2B 

Fish analysis Llv No.1 Commenls: 

Sample Type: Unknown Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC-20AO COLUMN : Hypersl GOLD 2.1 X 50 mm (5 um) ELUENT : A;20 mM Ammonium fonnale aqJform!o acid (1000:1) 

B; Acefonltrlla 

A!B(1:1) FLOW RATE : 0.2 ml!mln COLUMN TEMP : 313 K 

INJECT VOL : 5 ut 

"' ANALYTICAL CONDITIONS (~lS) '" 

MS TYPE : Thenno Fisher Sclentlllc TSQ Quantum Access MAX IONIZATION : Nega!lve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATHGAS : 00 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 100) 

''''" 

Data File: C186FA28L2 Aoqulslllon Date: 

Original Data Path: 

Sample Name: 

Sample Type: 

C:\Xclllibur\Dala\~aneko\C166\0a SampleiD: 

y2B 

Flsh anatys!s llv No.2 Commenfs: Unknown Ope-rator. 

'" At/AL YTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHII.'.ADZU LC·ZOAD COLUMN : HypersQ GOLD 2.1 x 50 rr.m (5 um) 

ELUENT : A; 20 mMAmmonlum formals aq.trom!le acid (1000:1) B;Aca!onitrllo 

AIB(1:1) 

FLOWRATE : 0.2mllmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' A~IAL YTICAL CONDITIONS (MS) m 

MS TYPE : Thermo Fl•her SdenUiic TSQ Quantum Access MAX IONIZATION : NegaUve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz180) 

= ·--u

66 
8-2 Control group .-------------------~ 

Fish analysis 

DayO 

Standard solution 

0.0048 mgL"1 

Fish analysis 

DayO 

Standard solution 

0.0964 mgL· 1 

Da!a Fife: C1BSWFAOS1 Acqulslt!on Dalo: Original Data Palh: C:\Xcallbur\Oa!a\kaneko\C166\0a Sample 10: yO 

Sampfo Name: Std. 0.0048 mgll. Commsnts: 

Sample Typo: Sid Bracket Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

SHIMADZU LC·20AD Hyper<B GOLD 2.1 x 50 mm (5 urn) 

A: 20 mM Ammonium formate aqlfonnfcacld (1000:1) B;Acetonlltile 

A/8(1:1) : 0.2mUmln : 313K : suL 

•~ ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Sclenlific TSQ Quanlum Access MAX IONIZATION : Negalive ESI 

VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (rnlz 1 80) 

................................................................................................................................... 

Dala File: C186WFAOS2·2 Acquisition Dale: Original Dala Palh: C:\Xcallliut\Dala\kaneko\C186\Da Sample 10: yo 

Sampfe Name: Std. 0.0964 mgJL Comment~: 

Sample Typa: StdBracket Operator: 

"'ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMAOZU LG-20AD 

: Hypersll GOLD 2.1 x 50 mm (5 um) : A; 20 mM Ammonium fonnale aqlfo;mlc acid (1000:1) 

B; Acelon!trlle NB(1:1) 

: 0.2 mllmln : 313 K : 5uL 

"' ANALYTICAL CCNDITIOtiS (MS) ••• 

MS TYPE : Thermo Flsher SclenUfic TSQ QuantUJ"' Access MAX IONIZATION : Nega!lvo ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 16 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

67 
Fish analysis 

DayO 

(before starting exposure) 

Control group 

Fish analysis 

Day28 

Standard solution 

0.0048 mgL"1 

DafaFtla: C185FAOC Acqulslllon Data: 

Original Oala Palh: C:\Xcalibuf\Dala\kanel<o\C186\0a Sampl< 10: 

SamptG Name: Sample Type: 

~~h analysis Ctrl Comments: Unknown Oporalor: 

........................................................................................... , 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

aUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMADZU LC.ZOAD : Hype!SII GOlD 2.1 x 50 mm (5 um) : A; 20 mM Ammonium l01111a!e eqJform!c acid (1000:1) 

B; Acelonllrile 

AIS(1:1) : 0.2mUmln : 313K 

: 5ul 

'"ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclantl!io TSQ Quanlum Accass MAX 

iONIZATION : Negallve ESI 

VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCANTYPE : SIM(mlz180) 

................................................................................................................................... 

""" """"

OataFne: C186WFA2BS1 AcqulsiUon Date: Original Data Path: 

Sampfe Name: 

Sampfo Type: 

C:\Xcalibur\Dala\kancl<o\C186\0a Sampla!O: 

y2a 

Std. 0.0048 mgJL Comments: Std Bratket Operator: 

"' ANALYTICAl COilCITIONS (HPLC) '" 

HPLC TYPE : SHIM .. OZU lC.20AO COlUMN : HypmUGOlD 2.1 x50mm(5um) 

ElUENT : A; 20 mM Ammcnlum f01111ala aqJformlc acid (1000:1) 

B;Acetonlllllo 

N8(1:1) 

FLOW RATE : 0.2 mtlmin COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAl CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Sc!en!ilic TSQ QuanttJJ11 Access MAX IONIZATION : Nega!lve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 

AUXGAS ! 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 160) 

68 
Fish analysis 

Day28 

Standard solution 

0.0964 mgL-1 

Fish analysis 

Day28 

(at the termination 

of exposure) 

Control group 

Data File: C186WFA28S2·2 Acquisition Dale: Original Data Path: C:1Xcal~u~Oalalkaneko\C1B6\Da Sampla!D: 

y2B 

Sampro Nama: 

sample Type: 

Std. 0.0964 mgll.. Comments: Sid Br.u:kel Operator: 

'" ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHlMADZU LC-20AO 

COLUMN : Hypersll GOLD 2.1 X 50 mm (5 um) ELUENT : A: 20 mM Ammonium rormalo aqJformlc acid (1000:1) 

B;Acetoni!tito 

AIB(1:1) FLOWRATE : 0.2m!lmln COLUMN TEMP : 313 K INJECT Vat. : 5 uL 

~• ANAlYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Asher Sdenlifio TSQ Quanlum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOI.TAGE : 2500V SHEATHGAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 180) 

Data Flto: C1B6FA28C Acquisition Date: 

Original Dala Palh: C:\Xcalibu~aio\kaneko\C185\Da Sample ID: 

y2B Sample NamG: Flsh analysis Ctrf Comments: 

Sample Typo: Unkno\vn Operator: 

'" ANAlYTICAl CONDITIONS (HPLC) '" 

HPlCTYPE 

COLUMN 

ELUENT 

SHIMADZU tC-20AO : Hypersil GOLD 2.1 x 50 mm (5 um) : A: 20 mM Ammonium forrnalo aq./formlo aold (1000:1) 

B; Acetonlltllo 

FLOW RATE COLUMN TEMP INJECT VOL 

AIB(1:1) : 0.2mlJmln : 313K : 5uL 

"' ANALYTICAL CONDITIONS (MS) "' 

MS lYPE : Thermo Fisher Sc!ontif'JOTSQ OuanlumAccess MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

69 
Reference 9 

9-1 

Chromatograms when measuring the concentrations in water 

Test group r-------------------~ 

Water analysis 

DayO 

Standard solution 

0.0048 mgL"1 

Water analysis 

DayO 

Standard solution 

0.0964 mgL"1 

Data File: 

Orlglnnl Data Path: 

Sample Name: Samplo Typo: 

C166WFAOS1 Acqulslllon Dale: C:\Xcallbw\Dalo\kaneko\C16o\Da SampteiD: 

yo 

Sid, 0.0048 rr.gll Comments: 

Sid Braeket Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHtMAOZU LC.20AO COlUMN : Hyparsi!GOLD 2.1x50mm(5um) 

ELUENT : A: 20 mM Ammonium formalo aq./formlo acid (1000:1) 

B; Aoetonlltila IVB(1:1) FLOW RATE : 0.2 mUmln COlUMN TEMP : 313 K INJECT VOL : 5 ul 

'" AIIALYTICAL CONDITIONS (MS) '" 

MS TYPE : Thermo Fisher Sclonlifr. TSO Quanlum Access MAX IONIZATION : NegatiVe ESI VAPDRIZERTEMP: 723K 

SPRAYVOlTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE ; SIM (mlz 180) 

RT:O.WJ~3.00 UI:1SG 

"'''"

OalaFife: 

Original Data Path: 

Sampla Name: Sample Type: 

C186WFAOS2·2 Acqulsl!lon Date: 

C:\XcaHbur\Dalalkaneka\C186\Ila SampleiD: 

yo 

Std. 0.0904 mgJL Commenls! 

Std Bracket Operator: 

••• AIIALYTlCAL CONDITIONS (HPLC) "' 

HPLCTYPE : SHIMAOZU LC.20AO COLUMN Hypersl! GOLD 2.1 x 50 mm (5 urn) ELUENT A; 20 mM Ammonium formate aq.lformlo acid (1000:1) 

B; Acetonilrlle Alll(1:1) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDITIONS (MS) ••• 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : llegallve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2SOOV SHEATH GAS : 50 AUXGAS ; 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAli TYPE : SIM (mlz 180) 

70 
Water analysis 

DayO 

High exposure level 

Water analysis 

DayO 

Low exposure level 

Data File: 

Original Data Path: 

Sample Name: 

Sampio Type: 

C186WAOH3 Acquls!Uon Date: C:\Xca!ibur\Dafa\kaneko\0186\0a SampleiD: 

yO 

Wale:r analysTs H Lv Comments: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC·20AD 

COLUMN : HypersllGDLD 2.1x50rrm(5um) ELUENT : A;20 mM Ammonium formate aqJformlo acid (1000:1) B; Acetonitrile A/8(1:1) FLOWRATE : 0.2mUmln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDmONS {MS) "' 

MS TYPE : Thermo F~her Scienlif!C TSQ Quanfum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS ! 50 

AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 180) 

Data File: C186WAOL Acquisition Dato: 

OrlglnaJ Data Path! C:\Xcnllbur\Oala\kaneko\C1 B6\Da Sample 10: 

yo Sample Name: Wafer analysis Ltv Comments: Sample Type: Unknown oP-arator. 

"' ANAL YnCAl CONOmONS {HPLC) ri• HPLC TYPE : SHIMADZU LC.2DAD COLUMN Hypl!tsD GOLD 2.1 x 50 mm {5 um) aUENT A; 20 mM Ammonium roll!lale aq./Tormlo ocld {1000:1) Bi Aeelonltdte 

A/8(1:1) FLOWRATE : 0.2ml!mln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

'" ANALYTICAL CONDITIONS {MS) '" 

MS TYPE : Thermo Fisher Sclenllfic TSQ Quantum Access MAX 

IONIZATION : Negallve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 1 BO) 

71 
Water analysis 

Day7 

Standard solution 

0.0048 mgL'1 

Water analysis 

Day7 

Standard solution 

0.0964 mgL'1 

Oala Fila: C166WFA7S1 Acqulslllon Dalo: 

Original Oala Palh: C:\Xcaf:lxu\Oala\l<llnekDIC166\Da SampleiD: 

y7 

Sample Namo: Std. 0.0048 mg/L Comments: Sampfe Typo: Std Bracl:et Operator. 

"' ANALYTICAL CONDITIONS (HPLC) ''' 

HPLC TYPE : SHIMADZU LC·20AD COLUMN : HypersU GCLD 2.1 X 50 mm (5 urn) ELUENT : A: 20 mM Ammonium formate aq.ffonnle add (100o:1) 

B; Acelonllrife N8(1:1) FLOW RATE : 0.2 mllmln CCLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTJCALCONDITIONS(MS) "' 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negallve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 !ONSWEEPGAS : 0 CAPILlARY TEMP : 493 K SCAN TYPE : SIM (mh: 180) 

DataFite: 

Orlglnar Data Path: 

Sample Name: 

Sample Type: 

C186WFA7S2 Acqulslllon Dale: 

C:\Xcau~ur\Dala\kanel<o\C1B8\Da SamploiD: 

y7 

Sfd. 0.0964 mg/L Comments: 

Std Bracket Operator: 

PO ANALYTICAL CONDITIONS (HPLC) -

HPLC TYPE : SHIMADZU LC-2DAD CCWMN : Hypersll GOLD 2.1 x 50 mm (5 um) ELUENr : A; 20 mMAmmonlom fomrate aqJform!c acld (1000:1) 

B;AcalonlliUe 

N8(1:1) FLOW RATE : 0.2 mUm!n COLUMN TEMP : 313 K 

INJECT VOL : 5 uL 

•u ANALYTJCALCONOITIONS (MS) '" 

MS n'PE : lhenno Ffsher Scientific TSQ Quantum Access MAX IONIZATION : Negatlve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V 

SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPillARY TEMP : 493 K SCANTYPE : SIM(m/z!BO) 

Jtr:II.LII.:f.M nt1!G 

"''" ,_, 

,,_. '""" 

f1oo:;ooo 

~ 

"""' 

"""· 

""" """" 

72 
Water analysis 

Day7 

High exposure level 

Water analysis 

Day7 

Low exposure level 

DalaFIIe: Ci SSWA7H Acqulslllcn Oate: 

Original Oafa Path: C:\Xcall00r\Data\l:aneko\C186\Da Sample!D: 

y7 

Sampla Nama: Water analysis H Lv Comments: Sample Type: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) «> 

HPlC TYPE : SHIMADZU LC·20AO COLUMN : HypersH GOLO 2.1 X 50 mm (5 um) ElUENr : A: 20 mMAmmentum rormefe aqJrormk:acld (1000:1) 

B;Acelonltrll-a 

AJB(1:1) FLOWRATE : 0.21nl/mln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

'" ANALYTICAL CONDITIONS (MS) '" 

MS TYPE : Tharmo Fisher SclenOITe TSQ Quantum Access MAX IONIZATION : Negative ESt VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARYTEMP : 493 K SCAN TYPE : SIM (mlz 180) 

DafaFife: 

Origfnal Data Path: C186WA7L 

C:\Xcalilror\Oafa\l:aneko\C186\Da AcqulsJUon Data: SampleiO: 

SampJo Name: 

Sample Type: ~!er ana~tsls llv Unknovm Comments: 

Operator: 

............................................................................................. "' ANALYTICAL CONDITIONS (HPLC) k• HPLO TYPE : SHIMADZU LC-20AD 

COLUMN : Hypersff GOLD 2.1 x 50 mm (5 um) 

ELUENT : A; 20 mM Ammonium ronmale aqJiormlc acid (1000:1) B;AcsfOfl«rita 

AJB(1:1l FLOW RATE : 0.2 mllm!n COLUMN TEMP : 313 K INJECT VOL : 5 UL 

"' AIIALYTICALCONOITIONS(MS)'" 

MS TYPE : Thanmo Flsi>er Sclentiffo TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCANTYPE : SIM(mll:150) 

................................................................................................................................... RT:OJIO•),Ill Si.t-UQ 

"'"" ,_, 

O:,

73 
Water analysis 

Day 14 

Standard solution 

0.0048 mgL"1 

Water analysis 

Day 14 

Standard solution 

0.0964 mgL"1 

Data File: C186WFA14S1 Aequlsfllon Date: Original Data Path: C:\Xcallbw\Oala\kaneko\C186\0a SampleiO: y 14 Sample Namo: Std. 0.0048 rng!L Comments: Sampfe Typo: Std Bracket Opera!or. 

'"ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE : SHIMADZU LC-20AD COLUMN : Hypers!GOLD 2.1x50mm(5um) 

ELUENT : A;20mMAmmorJumrormaloaq.lformicaeid(1000:1) 

8; Acelon~rlte 

AIB(1:1) FLOW RATE : 0.2 rntimln COLUMN TEMP : 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thenno Fisher Scfentlfic TSQ Quan!um Access t.-i.AX IONIZATION ! Negallve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V 

SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPilLARY TEMP : 493 K SCANTYPE : SIM(mlz1BO) 

__

Data File: C1B6WFA14S2 AcqulsiUon Dale: 

Original Data Path: C:\Xcalibur\Oala\kaneko\C18S\Oa SamploiD: 

y14 

Sample Nama: Std. 0.0964 mgll Comments: 

Sampre Typo: Std Bracket Operator. 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

: SHIMAOZU LG-20AD : Hypersl GOlD 2.1 x 50 mm (5 urn) : A; 20 mM Ammonium formate aq.lformlc acid {1000:1) 

B;AC<>!onl!n1e 

FLOW RATE COWMN TEMP InJECT VOL 

N8(1:1) : 0.2 m!Jmln : 313K 

: 5 uL 

"' ANALYTICAL CONDITIONS (MS} -

MS TYPE : Thermo fisher ScfenUfic TSQ Quenlum Aecass MAX 

IONIZATION : Nogatve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPilLARY TEMP : 493 K SCANTYPE : SIM(mlz18G) 

U~Q

~~=""" """ "'"'

""" 

74 
Water analysis 

Day 14 

High exposure level 

Water analysis 

Day 14 

Low exposure level 

Data Ftfe! C1B6WA14H Acquisition Dale: Original Dala Path: C:\Xcallbw\Dala\kaneko\0186\Da SampleJO: 

yt4 Sampfo Namo: Water analysis H lv Comments: 

Sample TYpo: Unknowl\ Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMAOZU LC-20AD COLUMN Hyporsl!GOLD 2.1x50mm(Sum) ElUENT A; 20 mM Ammonium form ale aq./formlc acld (1000:1) 

SjAce!onllrile 

AIB(l:l) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"' ANAL YTJCAL CONDinONS (MS) '" MS 1YPE : Thermo Flsher Sclentlflc TSQ Quantum Access MAX IONIZATION : Negallve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V 

SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 1 BO) 

--

Oala File: C1B6WA14L Aequlslllon Dale: Original Data Path: C:\Xcalibu!IOala\kanekoiC18610a Sample to: y14 Sample Namo: Water analysts L Lv Comments: 

Sample Type: Unknown Operator: 

'" ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE : SHIMADZU lC-20AD COLUMN : HypersU GOlD 2.1 X SO mm (5 urn) ELUENT : A; 20 mM Ammonium formate eqJIO!l11k: acld (1000:1) Bi AcelonllrUa AIB(1:1) FLOWRATE : 0.2mVmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclenlllio TSQ Qoanlom Access MAX IONIZATION ; Negative ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (mlz 1 80) 

"""'

75 
Water analysis 

Day21 

Standard solution 

0.0048 mgL"1 

Water analysis 

Day 21 

Standard solution 

0.0964 mgL·1 

Data File: C186WFA21S1 Acquisition Date: 

Original Data Path: C:IXcafibur\Dala\kanel<o1C1SS\Da Sampto to: 

Y21 Sampto Name: Std. 0.0048 mg/L Comments: 

Sample Typo: Sld Bracket Operator: 

"' ANALYTtCALCONDITIONS (HPLC) "' 

HPLC TYPE : SHIMAOZU LC-20AD COLUMN : Hypersll GOLD 2.1 x 50 mm (5 urn) ELUENT : A: 20 mM Ammonium fcrmele aqJroon!c acid (1000:1} Bi Acelanilrllo NS(1:1) FLOW RATE : 0.2 mUmln COLUMN TEMP : 313 K INJECT VOL : 5 Ul 

"' ANAL YTJCAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Scler.fltic TSQ Quantum Acass MAX IONIZATION : Negalive ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATHGAS : 50 AUXGAS • : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCANTYPE : SIM(mlz100) 

Data fila: C156WFA21S2 Acquisition Date: OrlglnaJ Dala Path: C:\Xcalibur\Oaia\keneko1C1S6\0a Samplo!D: 

y21 Sample Name: Std. 0,0964- mgll. Comments: 

Sample Typa: Std Bracl<et Operator: 

"*ANALYTICAL CONDITIONS (HPLC) '" 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMAOZU LC-20AO : Hyparsn GOLD 2.1 x 50 mm (5 um) 

A: 20 mMAmmonhnn formale oqJiormlc acid (1000:1) a; Acetonilrne 

NS(1:1) : 0.2mllmin : 313K : SuL 

.,, ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 60 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m!z 180) 

76 
Water analysis 

Day21 

High exposure level 

Water analysis 

Day21 

Low exposure level 

Data File: C186WA21H Acqulslllon Date: 

Original Data Path: C:\XcallbU1\0ala\kaneko\C188\Da SamplaiD: 

y21 

Sample Name: Waleranalysls H lv Comments: Sampro 1'ype: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLClYPe 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMADZU LC·20AD : HypersU GOLD 2.1 X 50 mm (5 um) 

: A; 20 mM Ammonium formate aq./formlc acid (1000:1) B; AcetonUnla A/8(1:1) 

: 0.2mUmln : 313K 

: 5 ul 

"' ANALY11CAL CONDITIONS (MS) "' 

MS TYPE ! Thermo Fisher Sclentinc TSQ Quan!um Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : o CAPILLARYTEMP : 493 K SCAN TYPE : SIM (m/z 180) 

Data File: Orlgtnal O;ata Path: 

Sample Name: 

Sample Typo: 

C186WA21L Acquisition Dale: 

C:IXcaL'bu~Oala\kaneko\C186\Da Sample 10: y21 

Water analysis L Lv Ccmmcnfs! 

Unl<nown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE : SHIMADZU LC·20AD COLUMN : HypersU GOLD 2.1 x 50 mm (5 urn) 

ELUENT : A; 20 mM Ammonium formate aqJformlc acid (1000:1) B; Acetonitrile Nl3(1:1) FLOW RATE : 0.2 mllmln COLUMN TEM? : 313 K INJECT VOL : 5 uL 

'" ANALYTICAL CONDITIONS (MS) '" 

MS TYPE; : Thanno Fisher Sclenllffa TSQ Quanlum Ac<:ess MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN lYPE : SIM (m/z 180) 

RT:ilctlo:l.£10 $J.t:14G 

' 

-------,c:::

77 
Water analysis 

Day23 

Standard solution 

0.0048 mgL"1 

Water analysis 

Day23 

Standard solution 

0.0964 mgL·1 

Data Ale: C186WFA23S1 Acquisition Dale: 

Original Data Pat~: C:\Xcallbu!\Dala\l<aneko\C18G\Oa SampleiO: 

y23 

Sample Name: Std. 0.0046 mgfl Comments: 

Sampto Type: Std Bracket Operator. 

'" ANALYTJCALCONDITIONS(HPLC) •~ 

HPLC lYPE : SHIMAOZU LC-20AO COLUMN : Hyporsn GOLD 2.1 X S<l mm (5 um) ELUENT : A; 20 mMAmmonlum formals aqJfennlc ncld {1000:1) B: Acetonlln1e NS(1:1) 

FLOWRATE : o.2m!.lmln COLUMri TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICAL CONDITIONS (MS) "' MS TYPE : Thermo Fisher Sclentiflo TSQ Quantum Access MAX ICNIZATKlN : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : :!SOOV SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARYTEMP : 493 K SCAN lYPE : SIM (mlz 180) 

U:!lOOO•

•,

___

Data Fila: C186WFA23S2 Acqulailfon Dale: 

Ortgtnal Data Path: 

Sample Namo: 

C:\XcafibUI\Oala\l<aneko\C186\Da SampleiD: 

y23 

Sid. 0.0954 mgll. Comments: Sample Typo: S!d Bmcket Operator: 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLC TYPE SHIMAOZU LC-20AD COLUMN Hypersa GOLD 2.1 x SO mm (5 um) 

ELUENT A; 20 mM Ammonium fonnalo aqlfO!f111c acld (1000:1) 

B; Acekm!~Ha 

A/8(1:1) FLOW RATE : 0.2 mUrn!n COLUMN TEMP : 313 K INJECT VOL : 5 ul 

... ANALYTICAL CONDITIONS (MS) ~ 

MS TYPE : Thermo Fisher Scientific TSQ Quantum Access MAX IONIZATION : NegaUve ESI VAPORIZER TEMP: 723K SPRAYVOLTAGE : 2SOOV SHEATH GAS : 50 AUXGAS : 15 IONSWEEPGAS : 0 CAPILLARYTEMP : 493K SCAN lYPE : SIM (m/z 160) 

""" o;+

--------

78 
Water analysis 

Day23 

High exposure level 

Water analysis 

Day23 

Low exposure level 

Dataflfe: 

Original Data Path: 

Sample Name: Sample Typo: 

Cf85WA23H 

C:\XcelbUJ\Oe!a\kaneko\C186\Da 

W~tr analyslsH Lv 

Unknown 

Aequisltlon Date: 

Sample 10: 

Comments: 

Operator: ............................................................................................ "' ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COlUMN TEMP INJECT VOL 

: SHIMAOZU lC·20AD : HypersD GOlO 2.1 x 50 mm (5 um) : A;20 mMAmmon!um formate aqJformlc acld{1000:1) B; Acelonlltile Ml{1:1) 

: 0.2mUmin 

: 313K : 5ul 

ou ANALYTICAL CONDITIONS (MS) -

MS TYPE : Thermo Asher Sclentmo TSQ Quan!um Access t..tAX IONIZATION : Noga!Ne ESI VAPORIZER TEMP ! 723 K SPRAYVOlTAGE : 2500V SHEATH GAS : 50 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE ! SIM (mlz 160) 

................................................................................................................................... 

""" "''''

"""

___

Data File: C166WA23L Aequlslllon Pate: 

Orlgtnal Dnla Path: C:\Xca!ibur\Oala\kaneko\C1 66\0a Sample 10: y23 

Sample Name: Waler analysis L lv Comments: Sampta Typt: Unknown Operator: 

"' ANALYTICAL CONDITIONS (HPLC) '" 

HPLC TYPE SHIMAOZU lC-20AO COlUMN HypeiSR GOLD 2.1x 50 rnm (5 urn) ELUENT A; 20 mM Ammonium fonnalo aqJformlc sold (1000:1) B; AcetonltrUa 

FlOWRATE : ~{~Bmln COlUMN TEMP : 313 K INJECT VOL : 5 uL 

•~ ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thenno Fisher Sclannnc TSQ Quantum Access MAX IONIZATION : NegaUva ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 25COV SHEATHGAS : 5I) 

AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : S!M (mlz 180) 

flf;O.W·~$1.1:1~0 

. 

79 
Water analysis 

Day28 

Standard solution 

0.0048 mgL"1 

Water analysis 

Day28 

Standard solution 

0.0964 mgL"1 

Data File: 

Origrnal Data Path: C1BGWFA2BS1 Acquisition Date: 

C:IXcallbUIIOala\kanako\C1B6\Da Sample ID: y29 Sample Name: Std. 0.0048 mg/L Comments: SamplaT)'pe: Sid llmket Operator. ........................................................................................... "' ANAL YTfCAL COI>IOJTIONS (HPLC) "' 

HPLCTYP~ 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

SHIMADZU LC.2ilAO 

Hypersl GOLD 2.1 x 50 mm (5 urn) 

A; 20 mMAmmoolum formaloaqJformlc acid (1000:1) B;AcelonlltHe 

Al!l(1:1) : 0.2mUmln : 313K : 5ul 

"' ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Scfenlific TSQ Quantum Access MAX IONIZATION : Nega!We ESI VAPORIZER TEMP: 723K 

SPRAYVOLTAGE : 2500V 

SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 

CAPILLARY TEMP : 493 K SCAN TYPE : SIM (rrk 180) 

................................................................................................................................... 

""" 0 

.. ___

Data File: C1BoWFA28S2·2 AcqulsfUon Date: 

Original Data Path: C:\XcaUbur\Oata\kaneko\C165\0a Sample JO: 

y2B Sample Name: Std. 0.{)964 mgJ1. Comments: 

Sampf& Type: Sid Bracket Operator. 

"' ANALYTICAL CONDITIONS (HPLC) "' 

HPLCTYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP 

INJECT VOL 

•~ ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thenno Fisher Sd&ntffic TSQ Quantum Access MAX IONIZATION : Nesallve ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (ml< 180) 

..

80 
Water analysis 

Day28 

High exposure level 

Water analysis 

Day28 

Low exposure level 

Data File: C185WA26H Acquisition Date: Original Dala Path: C:\Xcallbu11Data\kaneko\C165\Da SamploiD: 

y28 

Sample Name: Water analysis H Lv Comments: 

Sample Type: Unknown Oporator: 

'" ANALYTICALCONDITIOilS(HPLC) "' 

HPLC TYPE : SHIMADZU LC-20AD COLUMil : Hypersll GOLD 2.1 X 50 mm (5 um) ELUENT : A; 20 mM Ammonium formal• aqJfotmlc acid (1000:1) 

B; Ac:etonltn1a 

FLOW RATE : ~(~Umln COLUMN TEMP : 313 K INJECT VOL : 5 ut 

•u ANALYTICAL CONDITIONS (MS) •u 

MS lYPE : Thermo Fisher Scienllfic TSQ Quantum Access MAX IONIZATION : Nogallvo ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K 

SCAN TYPE : SIM (11'1Z 180) 

Data Fllo: C166WA28t Acquisition Date: 

Orfglnaf Oata Path: C:\XcaffburiDala\lcane!:olC1S6\0a Sampte JD: y26 

Sample Name: Water anafysls l Lv Comments: Sample Type: Unknown Operator: 

u• ANAL YilCA!.. CONDITIONS (HPLC) u• 

HPLC TYPE : SHIMADZU LC-20AD COLUMN H1Jl'!I>U GOLD 2.1 x 50 mm (5 um) ELUENT A; 20 mMAmmonltJm formate aqJformla ackl (1000:1) 

B;Acelonl!rlle 

A/!!(1:1) FLOWRATE : 0.2mUmln COLUMN TEMP : 313 K INJECT VOL : 5 ul 

HO ANAL YT!CAL CONDITIONS (MS) '" 

MS lYPE : Thermo Fisher Sclenfirlc TSQ Quantum Access MAX 

IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V SHEATHGAS : 50 AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN TYPE : SIM (m/z 160) 

Rr:;.oo.:t..OO tJ.t1SG 

·=

'"""' 

~.a

81 
9-2 Control group .---------------, 

Water analysis 

DayO 

Standard solution 

0.0048 mgL"1 

Water analysis 

DayO 

Standard solution 

0.0964 mgL"1 

Data Ale: C1S6WFAOS1 Acqulsl!lcn Dala: Original Dala Path: C:\Xcalibur\Dala\kanekc\C1B6\0a SamplaiD: 

yo Samplo Name: Std. 0.0048 mg/L Comments: 

Samplo Typo: Sid Bracket Operator: 

"'ANALYTICAL CONDITIONS (HPLC) ~· 

HPLOIYPE 

COLUMN 

ELUENT 

FLDIVRATE 

COLUMN TEMi' INJECT VOL 

: SHIMADZU l0-20AD : Hypar.dl GOlD 2.1 X 50 mm (5 um) : A: 20 mM Ammonium fcrmala aqJfcrmlc acid (1000:1) B; Ace!onilllla AIB(1:1) : 0.2mllmlll : 313 K : Sci. 

"' ANALYTICAL CONDITIONS (MS) ••• 

MS TYPE : Thermo Fisher Sdenti!ic TSQ Quantum Access MAX IONIZATION : Negative ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500V 

SHEATH GAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPILLARY TEMP : 493 K SCAN IYPE : SIM (mlz 160) 

'"""' '"'"' 

c'

Data Fila: C1e6WFAOS2·2 Acquisition Da1o: 

Original Data Path: C:\Xcallbur\Dal2\kane~o\C186\0a SamploiD: 

yO 

Sample Name: Sid. 0.0964 mg1L Comments~ 

Sample Type; Std Bracket Operator. 

"' ANALYTICAL CONDITlONS (HPLC) "' 

HPLCIYPE 

COLUMN 

ELUENT 

FLOW RATE COLUMN TEMP INJECT VOL 

: SHIMADZU LC·20AD : Hypetsll GOLD 2.1 x 50 mm (5 um) A: 20 mM Ammonium fCimolo aq.fformlc acid (1000:1) 

B;Ace!onltriJe 

IVB(1:1) : 0.2mtlmln : 313K : 5ul 

'" ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Th<nr.c Fbher SclenUfc TSQ Ouanlum Access MAX IONIZATION : Nega!lvo ESI VAPORIZER TEMP : 723 K SPRAYVOLTAGE : 2500 V SHEATH GAS : 50 AUXGAS : 15 ION SWEE? GAS : 0 CAPILLARY TEMP : 493 K SCAN IYPE : SJM (mlz 180) 

82 
Water analysis 

DayO 

Control group 

Data Fife: C186WAOC1 AcqulsiUon Dalll: 

OtJginaJ Dafa Path: C:\Xcatihur\Oala\kaneko\C1BS\Da Sample 10: yo Sample Nama: Water analysts Ctrt Comments: 

Samplo Typo: Unknown Operator: 

H. ANALYTICAL CONDITIONS (HPLCJ ... 

HPLC TYPE SHIMAOZU LC-20AD COLUMN Hypers» GOLD 2.1 x 50 mm (5 urn) 

ElUENT A; 20 mMAmmonlum formal• aq.Jrormla acid (1000:1) B; Acelonilrife A/8(1:1) FLOW RATE : 0.2 mUmin COLUMN TEMP : 313 K INJECT VOL : 5 uL 

"' ANALYTICAL CONDITIONS (MS) "' 

MS TYPE : Thermo Fisher Scfenllfio TSQ Quanlum Access MAX IONIZATION : Negal!ve ESI VAPORIZER TEMP : 723 K SPRAY VOLTAGE : 2500 V 

SHEATH GAS : SO AUXGAS : 15 ION SWEEP GAS : 0 CAPiLLARY TEMP : 493 K 

SCAN TYPE : SIM (m'z 150) 

<>3

83 
Reference 10 Water quality test 

Analyzed at Institute of Ecotoxicology Co., Ltd. 

<Quantitative analysis of the residual chlorine> 

The quantification of the residual chlorine in water used in this study was performed 

according to JIS K0102 33.1 (a-Tolidine colorimetric method). 

The concentrations of the residual chlorine during the experimental period were 

determined to be 0.01 mgL- 1 or less. 

Concentration of residual Date of quantitative chlorine 

(mgL-1

) analysis 

<0.01 1.26.2011 

<0.01 2. 2.2011 

<0.01 2. 9.2011 

<0.01 2.16.2011 

<0.01 2.23.2011 

<Quantitative analysis of copper> 

The quantification of copper in water used in this study was perfonned according to ns KO 101 

51.1 (Absorptiometry). The concentration of copper was confinned to be 0.005 mgL-1 or less. 

Copper (mgL-1

) Date of quantitative analysis 

< 0.005 6.20.2010 

<Quantitative analysis of dissolved organic carbon concentrations> 

The quantitative analysis was performed using a total organic carbon meter TOC -

5000A (Shimadzu Corporation) (SOP/INS/4.9). 

The dissolved organic carbon concentration was confirmed to be 2 mgL-1 or less. 

Dissolved organic carbon 

concentration (mgL -

1

) 

<2 

Date of quantitative analysis 

2.14.2011 

84 
No. 106020 

April16,2010 

Report of the test results 

To the Institute ofEcotoxicology Inc. 

3-15-1 Komazawa, Setagaya-ku, Tokyo 

Shinnihon Environmental Research Corporation 

(Registration of Measurement Certification Business, 

Tokyo, No. 882) 

Akira Kishi, Executive Director 

The results of the test conducted on the sample as requested on April1, 2010 were as follows. 

Subject: Analysis of water used for raising carp 

Organization which Institute ofEcotoxicology, Inc. 

collected the sample: 

Type of sample: 

Date when sample was 

collected: 

Water quality 

April 1, 2010 

~ Water Quantifiable 

used for lower limit 

raising 

d carp 

Nitric nitrogen and (mg!L) 2.2 0.02 

nitrous nitrogen 

Potassium (mg!L) 1.5 0.5 

permanganate 

consumed 

Cyan (mg!L) Not 0.001 

detected* 

Total mercury (mg!L) <0.0005 0.0005 

Organic phosphorous (mg!L) Not 0.05 

detected* 

Iron (mg!L) <0.001 0.001 

Manganese (mg!L) <0.001 0.001 

Zinc (mg!L) 0.014 0.001 

Lead (mg!L) <0.001 0.001 

Hexavalent (mg!L) <0.005 0.005 

chromium 

Cadmium (mg!L) <0.001 0.001 

Arsenic (mg!L) <0.001 0.001 

Fluorine (mg!L) 0.10 0.05 

Total hardness (mg!L) 63 0.02 

Phenols (mg!L) <0.005 0.005 

Anionic surfactant (mg!L) <0.02 0.02 

pH( ) indicating water 7.4 0.1 

temperature at the time of (21.1) 

analysis 

Test methods 

Service water test method (2001) VI-2 13.2 

Service water test method (200 1) VI-1 17.2 

Service water test method (200 1) VI-2 22.5 

Service water test method (200 1) VI-3 25.2 

1974 Environmental Agency Notification No. 

64 Appendix 1 

Service water test method (2001) VI-3 13.4 

Service water test method (200 1) VI-3 12.4 

Service water test method (2001) VI-3 16.4 

Service water test method (2001) VI-3 27.2 

Service water test method (200 1) VI-3 11. 

Reference 2 

Service water test method (2001) VI-3 21.4 

Service water test method (2001) VI-3 17.2.2 

Service water test method (200 1) VI-2 3.2 

Service water test method (200 1) VI-I 15 .2.3 

Service water test method (200 1) VI-4 10.5 

Service water test method (200 1) VI-4 11.2 

Service water test method (200 1) VI-1 9.2 

* "Not detected" means when measured by the analytical method hsted above, the result was less than 

the quantifiable limit. 

85 
00 

0\ 

Reference 11 Records of observations of the conditions of the test fish (carp) 

Records of observations of the 

conditions of the test fish 

No.1 

Date Feeding Concentration Level I group 

period (Water tank No. 9) 

(days) Swim Food Health Water Mortality 

2011 status intake flow 

1.26 0 O.K. - O.K. O.K. 0 

1.27 I O.K. O.K. O.K. O.K. 0 

1.28 2 O.K. O.K. O.K. O.K. 0 

1.29 3 O.K. O.K. O.K. O.K. 0 

1.30 4 O.K. O.K. O.K. O.K. 0 

1.31 5 O.K. O.K. O.K. O.K. 0 

2.1 6 O.K. - O.K. O.K. 0 

2.2 7 O.K. O.K. O.K. O.K. 0 

2.3 8 O.K. O.K. O.K. O.K. 0 

2.4 9 O.K. O.K. O.K. O.K. 0 

2.5 10 O.K. O.K. O.K. O.K. 0 

2.6 II O.K. O.K. O.K. O.K. 0 

2.7 12 O.K. O.K. O.K. O.K. 0 

2.8 13 O.K. - O.K. O.K. 0 

2.9 14 O.K. O.K. O.K. O.K. 0 

Test substance 

Concentration Level 2 group 

(Water tank No. 8) 

Swim Food Health Water Mortality 

status intake flow 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

~ Flow rate: Level I: 300 ml/min, Lcvel2: 300 ml/min, Control: 300 ml/min to be confirmed. 

Swim 

status 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

~ Food intake: 0.050g per fish ~ Fcedprovided: Swimmy_(Japan Pel Food Col]l_., Lot No. 20 13.09.E3) 

~Cumulative mortality at the end of the test: Level I: 0%, Levcl2: 0%, Control: 0% 

~Remarks (Situation at the time of abnormality and treatments taken): Not applicable 

Control group 

(Water tank No. 7) 

Food Health Water 

intake flow 

- O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

- O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

O.K. O.K. O.K. 

- O.K. O.K. 

O.K. O.K. O.K. 

Test No. C-186 

Temperature/°C Observed 

by 

Mortality Level Level Control 

I 2 

0 24.8 25.1 25.5 Kaneko 

0 25.1 25.5 25.6 Kaneko 

0 252 25.3 25.5 Kaneko 

0 25.3 25.3 25.5 Kaneko 

0 25.5 25.3 25.5 Kaneko 

0 25.4 25.1 25.5 Kaneko 

0 25.5 25.3 25.5 Kaneko 

0 25.4 25.2 25.3 Kaneko 

0 25.5 25.5 25.3 Kaneko 

0 25.5 25.4 25.3 Kaneko 

0 25.5 25.4 25.5 Kaneko 

0 25.5 25.4 25.5 Kaneko 

0 25.3 25.5 25.5 Kaneko 

0 25.5 25.5 25.5 Kaneko 

0 25.5 25.5 25.4 Kaneko 
00 

.....:) 

Records of observations ofthe 

conditions of the test fish 

No.2 

Date Feeding Concentration Level 1 group 

period (Water tank No. 9) 

(days) Swim Food Health Water Mortality 

2011 status intake flow 

2.10 15 O.K. O.K. O.K. O.K. 0 

2.11 16 O.K. O.K. O.K. O.K. 0 

2.12 17 O.K. O.K. O.K. O.K. 0 

2.13 18 O.K. O.K. O.K. O.K. 0 

2.14 19 O.K. O.K. O.K. O.K. 0 

2.15 20 O.K. - O.K. O.K. 0 

2.16 21 O.K. O.K. O.K. O.K. 0 

2.17 22 O.K. - O.K. O.K. 0 

2.18 23 O.K. O.K. O.K. O.K. 0 

2.19 24 O.K. O.K. O.K. O.K. 0 

2.20 25 O.K. O.K. O.K. O.K. 0 

2.21 26 O.K. O.K. O.K. O.K. 0 

2.22 27 O.K. - O.K. O.K. 0 

2.23 28 O.K. O.K. O.K. O.K. 0 

Test substance 

Concentration Level2 group 

_(Water tank No.8) 

Swim Food Health Water Mortality 

status intake flow 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

O.K. - O.K. O.K. 0 

O.K. O.K. O.K. O.K. 0 

~ Flow rate: Level I: 300 ml/min, Level 2: 300 mllmin, Control: 300 mllmin to be confirmed. 

Swim 

status 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

O.K. 

~ Food intake: 0.050 g per fish ~Feed provided: Swimmy (Japan Pet Food Corp., Lot No. 20 13.09.E3) 

~ Cumulative mortality at the end of the test: Level 1: 0%, Level 2: 0%, Control: 0% 

~Remarks (Situation at the time of abnormality and treatrnenl~ taken): Not applicable 

Test No. C-186 

Control group Tempcraturef'C Observed 

(Water tank No.7) by 

Food Health Water Mortality Level Level Control intake flow 1 2 

O.K. O.K. O.K. 0 25.3 25.6 25.5 Kaneko 

O.K. O.K. O.K. 0 25.5 25.6 25.5 Kaneko 

O.K. O.K. O.K. 0 25.5 25.5 25.5 Kaneko 

O.K. O.K. O.K. 0 25.5 25.4 25.5 Kaneko 

O.K. O.K. O.K. 0 25.5 25.5 25.4 Kaneko 

- O.K. O.K. 0 25.5 25.6 25.4 Kaneko 

O.K. O.K. O.K. 0 25.1 25.4 25.1 Kaneko 

- O.K. O.K. 0 25.1 25.4 25.5 Kaneko 

O.K. O.K. O.K. 0 25.4 25.4 25.4 Kaneko 

O.K. O.K. O.K. 0 25.4 25.6 25.3 Kaneko 

O.K. O.K. O.K. 0 25.3 25.5 25.3 Kaneko 

O.K. O.K. O.K. 0 25.5 25.5 25.3 Kaneko 

- O.K. O.K. 0 25.3 25.5 25.2 Kaneko 

O.K. O.K. O.K. 0 25.2 25.1 25.3 Kaneko 
00 

00 

lt~rian t-rr,~-:.~~znuT'l'"'fl''ln~ 

l'l 

c 

2 

.id 

E 

~ 

!!! 

1-

~ 

)_ Namcofthctestsubstance: 

2. Lot No.: 

3. Date of measurement: 

4. Operator: Yasunori Okumura 

~ 

~ ~ 

~ 

= t':) 

(!) 

,.... 

~ 

II.>~ 

"'0 ~ (!) II.> 

a"'' .., (!) 

= t':) !3 ::;-

"'0 = 6 !3 

< ~ Q:""~ 

(!) """ Q..=" (!) 

~~ """II.> =-""" (!) II.> 

II.> = "'00"' 

0 ;!4. 

= ~ II.> 0 = t':) 

..:::;.,(!) 
IR spectrum (at the beginning of the study) measured at our location 

52.5 

45 

37.5 

30 

22.5 

15 

7.5 

-7.5 

4ooo 3aoo 3ioo 2s'oo 24oo 2000 1e'oo 16oo 

[C-186] 

File Name:c186s5 Sample: [C-186] 

Date/Time: 2010/12/03 14:43:06 

Type: IRPrestige-21 

Units: 1/cm 

Min: 399.26 

No. of Datapoints: 1868 

Gain: auto 

2 

3 

4 

5 

6 

7 

8 

No. of Scans: 20 

Ordinate: %T 

Max: 4000.36 

Data Interval: 1.93 

Aperture: auto 

89 

I .... I .... l. I'., .... I' I I I I 

1400 1200 1000 800 600 400 

Operator: 

Method: KBr 

Detector: standard 

Apodization: Happ-Genzel 

Resolution: 4 [1 I em] 

SpeedStd: 2.8 

1/cm 
IR spectrum (at the end of the study) measured at our location 

30 

22.5 

•• I I .. I. I' I I I I' I I I I I I I I I' I I I I j It I I I J I I I' I I I l' I I,-, I l-,, ~ ~ ~ ~ ~ ~ 1B 1B 1~ 1~ 1~ D B ~ 

[0-186] 1/cm 

File Name:c186e1 Sample: [C-186; 

Date/Time: 2011/02/25 9:39:17 

Type: IRPrestige-21 

Units: 1/cm 

Min: 399.27 

No. of Datapoints: 3735 

Gain: auto 

1 

2 

3 

4 

5 

6 

7 

8 

No. of Scans: 20 

Ordinate: %T 

Max: 400o.43 

Data Interval: 0.96 

Aperture: auto 

90 

Operator: 

Method: 

Detector: standard 

Apodization: Happ-Genzel 

Resolution: 2 [11om] 

SpeedStd: 2.8