REPORT

DETERMINATION OF THE PARTICLE SIZE DISTRIBUTION

OF TKA 40278 (CGI 827)

Data Requirement / Test Guidelines:

European Commission, ECB/TM/February 1996

Study Director:

Mrs. Alessandra Tognucci

Stud Corn letion Date:

March 07, 2003

Test Facility:

RCC Ltd

Environmental Chemistry & Pharmanalytics

CH-4452 Itingen/Switzerland

ClBA SPECIALTY CHEMICALS INC.

CH - 4002 Basel/Switzerland

RCC Stud Number:

845429

Page I of 16

272
RCC Study Number 845429 REPORT Page 2

TKA 40278 (CGI 827)

GLP CERTIFICATE

The Swiss GLP Monitoring Authorities

 ̄ Statement of GLP Compliance

It is hereby confirmed that

dudng the pedod of

the following Test Facilities of

August15-17,2000

-.August-28-29 ̄,2001 and.

.... Apd115,2002 ’ "

..... RCC Ltd .....

4452 Itingen

Switzerland

were Inspected by the Federal Office of Public Health, the Swiss Agency for the

Environment, Forests and Landscape and the Intercentonal Office for the Control of

Medicines with respect to the compliance with the Swiss legislation on Good Laboratory

Practice.

Test Facilities areas of expertise*

- Toxicology Division

- Environmental Chemistry and

Pharmsnalytics Diyis|on

- Microbiological Diagnostics by

Biotechnology & Animal Breeding Division

TOX, ACC, MUT

ACC, ECT, ENF, EMN,

PCT, RES,OTH (Animal

metabolism)

OTH (Microbiology)

The inspection was performed in agreement with the OECD Guidelines ~or National GLP

Inspections and Audits. It was found that the aforementioned test facilities were operating tn

compliance with the Swiss Ordinance relat ng to Good Laboratory Practice [RS 813.016.5] at

the time they were Inspected.

Federat Office of Public Health

The Director

Bern, May 2002 Prof. Th. Zeitner

 ̄ TOX = Tox~cok~y ; ACC = Analy~ca} end Clinical Chernlsky ; ECT = Environmental toxlc~ on ~quat~c and teuestdal

273
RCC Study Number 845429 REPORT Page 8

TKA 40278 (CGI 827)

GOOD LABORATORY PRACTICE

STATEMENT OF COMPLIANCE

RCC Study Number:

Test Item:

Study Director:

Title:

845429

TKA 40278 (CGI 827)

Mrs. Alessandra Tognucci

Determination of the Particle

of TKA 40278 (CGI 827)

Size Distribution

This study was conducted in compliance with the Swiss Ordinance relating to Good

Laboratory Practice, adopted February 2r~, 2000 [RS 813.016.5]. This Ordinance is based on

the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted November

26th, 1997 by decision of the OECD Council [C(97) 186/Final].

The purity of the test item is excluded from the statement of compliance (purity available in

Analytical Certificate No. 14026512-1; Ciba Specialty Chemicals Inc.).

Study Director: Tognucci

274
RCC Study Number 845429 REPORT Page 4

TKA 40278 (CGI 827)

SIGNATURES

Study Director:

Management: Dr. JSrn SchreitmQller

275
Rcc Study Number 845429 REPORT Page 5

TKA 40278 (CGI 827)

QUALITY ASSURANCE UNIT

RCC Ltd, Environmental Chemistry & Pharmanalytics, CH-4452 Itingen / Switzerland

STATEMENT

RCC Study Number:

Test Item:

Study Director:

Title:

845429

TKA 40278 (CGI 827)

Mrs. Alessandra Tognucci

Determination of the

of TKA 40278 (CGI 827)

Particle Size Distribution

The general facilities and activities are inspected periodically and the results are reported to

the responsible person and the management.

Study procedures were periodically inspected. The study plan and this report were audited by

the Quality Assurance Unit. The dates are given below:

Dates and Types of QAU Inspections Dates of Reports to the Study

Director and to the Management

September 16, 2002 Study Plan September 16, 2002

January 29, 2003 Process based January 29, 2003

(Measurement)

February 28, 2003 Final report February 28, 2003

This statement also confirms that this final report reflects the raw data.

Quality Assurance: Mrs. Diana B(~rgin

276
RCC Study Number 845429 REPORT Page 6

TKA 40278 (CG1827)

CERTIFICATION OF GLP

AND VERIFICATION OF THE REPORT

The statement of Compliance with Good Laboratory Practice found in this report, and signed

by the Study Director is truthful and accurate, and this report as provided by the testing

facility is complete and unaltered.

Signature of the Sponsor:

Date:

277
RCC Study Number 845429 REPORT Page 7

_TKA 40278 (CGI.___.827)

CONTENTS

.2

GLP CERTIFICATE ..............................................................

STATEMENT OF COMPL ANCE ................................. " ....................3

4

SIGNATURES ...................................................... ’ ’"

QUALITY ASSURANCE UN T .....................................................................

CERTIFICATION OF GLP ........................................................

6

AND VERIFICATION OF THE REPORT ....................................................................

CONTENTS ...............................................................

PREFACE ............................................................. " ..........................

8

9

DATA REQUIREMENT / TEST GUIDELINES .............................................

SUMMARY OF STUDY PLAN AMENDMENTS ..............................................................

9

1 PURPOSE ...........................................................................................................

10

2 MATERIAL AND METHODS ................................................................................

10

2.1 TEST TEM ............ " ...... 10

2.2 PRINCIPLE OF THE TEST ........................................................................

10

2.3 PERFORMANCE OF THE TEST.....~ ..........................................................

11

2.3.1 V~sua Exam nat on .....................................

11

..11

2.3.2 Test System ............................................................

2.3.3 Preliminary Study .......................................................................... 11

2.3.3 Main Study .................................................................................... 11

2.4 DATA HANDLING ......................................................................................

12

3 RESULTS .................................................. ’ ...........................

13

Table 1: Particle size distribution of TKA 40278 (CGI 827) .......................................

14

Table 2: Particle size distribution of TKA 40278 (CGI 827) in % at the relevant

partcesze .......................................

Figure 1 : Particle size distribution curve of TKA 40278 (CGI 827) (1. Run) ............... 16

278
RCC Study Number 845429 REPORT Page 8

TKA 40278 (CGI 827)

PREFACE

GENERAL

Title:

Sponsor:

Monitoring Scientist:

Test Facility:

Determination of the Particle Size Distribution

of TKA 40278 (CGI 827)

CIBA SPECIALTY CHEMICALS iNC.

CH-4002 Basel/Switzerland

Dr. Moung Sook Lee

RCC Ltd

Environmental Chemistry & Pharmanalytics

Zelgliweg 1

CH-4452 Itingen!Switzerland

RESPONSIBILITIES

Study Director:

Deputy Study Director:

Technical Coordinator:

Reporting:

Head of RCC Quality Assurance:

Mrs. Alessandra Tognucci

Dr. Ute Schmiedel

Mrs. Jos~e Gustaw, Mr. Christoph Wild

Mrs. Miriam Weissenfeld

Mrs. Iris WOthrich

SCHEDULE

Experimental starting date:

Experimental completion date:

Study completion date:

January 23,2003

January 24,2003

March 07,2003/wei

ARCHIVING

RCC Ltd, CH-4452 Itingen/Switzerland will retain the study plan, raw data, a sample of test

item and the final report of the present study for at least ten years.

No data will be discarded without the sponsor’s consent.

279
RCC Study Number 845429 REPORT Page 9

TKA 40278 (CGI 827)

DATA REQUIREMENT / TEST GUIDELINES

This study followed the procedures indicated by the following internationally accepted

guidelines and recommendations:

EUROPEAN COMMISSION, Directorate General XII-JRC, Science Research and

Development-Joint Research Centre. "Particle Size Distribution, Fibre Length and Diameter

Distribution" Guidance Document, ECBFFM/February 1996.

SUMMARY OF STUDY PLAN AMENDMENTS

First Amendment to Study Plan:

Concerning:

Alteration:

Reason for Alteration:

Schedule

New experimental starting and completion date

Delay in study

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RCC Study Number 845429 REPORT Page 10

TKA 40278 (CGI 827)

1 PURPOSE

The purpose of the study was to determine the particle size distribution of TKA 40278 (CGI

827).

2 MATERIAL AND METHODS

2.1 TEST ITEM

Data of test item as supplied by the sponsor.

Identity:

Product Name:

Batch:

Purity:

Appearance:

Expiration date:

Storage:

TKA 40278

CGI 827

KL 01/02

See Analytical Certificate, Study No. 14026512-1

Ciba Specialty Chemicals Inc

Solid, slightly yellow

August 30, 2004

At room temperature at about 20 °C.

The test item is light sensitive.

2.2 PRINCIPLE OF THE TEST

The above mentioned guideline contains several different methods for the determination of

the particle size distribution. These methods include sieving, microscopic examination,

sedimentation, gravitational settling examination, cascade impaction and laser scattering /

diffraction method. In the present study the laser scattering / diffraction was applied.

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RCC Study Number 845429 REPORT Page 11

TKA 40278 (CGI 827)

2.3 PERFORMANCE OF THE TEST

2.3.1 Visual Examination

Visual examinations were performed macroscopically and microscopically under

magnification of 400, using a light microscope LEITZ, DIALUX 20, to judge the colour and the

structure of the test item. For the examination under the microscope, the test item was

dispersed in white spirit containing Sympatens SHO/400, placed on a slide and covered with

a second slide. Details are part of the raw data and are given later in this report.

2.3.2 Test System

Particle size analyser

Instrument:

Particle size range:

Test item dispersion:

"Analysette 22" COMPACT, Fritsch GmbH using a micro

dispersing apparatus. The laser diffraction apparatus was

checked at the beginning of the measurement using a Fritsch

standard sample. The results measured are within the limits

given by Fritsch. Details are not reported here but are part of the

raw data.

0.3 - 300 pm

The test item was dispersed in white spirit containing

Sympatens SHO/400 at room temperature. The mixture was

ultrasonificated for 15 minutes before use.

2.3.3 Preliminary Study

A solubility test was performed to choose a suitable dispersing agent. For this test it is

required that the test item be both: sparingly soluble and well dispersible in the solvent.

During the solubility test, white spirit containing Sympatens SHO/400 was selected as an

appropriate dispersing agent for this study. A pre-saturated test item solution was prepared

by transferring 0.20 g of the test item to an Erlenmeyer flask.by bringing up the volume to 1 I

with white spirit and adding one drop Sympatens SHO/400. This mixture was stirred for

2 hours at room temperature and filtered with a 0.45 pm filter before use. The pre-saturated

white spirit solution was used to prepare a stock dispersion (0.2 g of the test item were mixed

with 10 ml pre-saturated white spirit) and for the measurement of the background of the laser

diffraction apparatus.

2.3.3 Main Study

The dispersing apparatus was rinsed and filled with pre-saturated white spirit. A few ml of the

stock dispersion were added slowly until the absorption of the laser was reduced by about

10 %. The particle size distribution in the suspension was quantified by laser diffraction. The

test was performed with two samples of the stock dispersion and each measured in

duplicate. Details are part of the raw data and are summarised in Table 1 and 2.

282
RCC Study Number 845429 REPORT Page 12

TKA 40278 (CGI 827)

2.4 DATA HANDLING

A particle size distribution curve was prepared in the range between 0.3 I~m and 300 pm. The

mass median diameter (MMD) of TKA 40278 (CGI 827), which is the particle size (pm), not

exceeded by 50 % of the particles was calculated. In other words, the size of 50 % of the

particles are smaller than the determined value and also, that 50 % of the total sample mass

is presented in particles having a diameter larger than the determined value.

283
RCC Study Number 845429 REPORT Page 13

TKA 40278 (CGI 827)

3 RESULTS

The determination of the particle size distribution of TKA 40278 (CGI 827) was performed

according to the European Commission, Document ECB/TM/February 1996: "Particle Size

Distribution, Fibre Length and Diameter Distribution" Guidance Document using the laser

diffraction method.

TKA 40278 (CGI 827) is a light beige granular powder. Under the microscope (with a

magnification of 400) elongated, squared, colourless, crystalline structures were observed.

A solubility test showed, that the test item is both: sparingly soluble and well dispersible in

white spirit containing Sympatens SHO/400. The particle size distribution of TKA 40278 (CGI

827) was performed with two samples of the stock dispersion one measured in duplicate.

The individual measurements and the results obtained using the laser diffraction of TKA

40278 (CGI 827) are summarised in Table 1 and Table 2. The tables represent rounded

results obtained by using the exact measurement data. An example of the particle size

distribution curve is shown in Figure 1.

The results obtained during the measurements showed no relevant difference. The MMD

mean value of 16.8 pm with a standard deviation of + 0.05 pm is given in Table 1. The

standard deviation is smaller than 5 % therefore the study is considered to be valid and no

further testing is necessary.

In conclusion using alaser diffraction apparatus to study the particle size of TKA 40278 (CGI

827) in the range between 0.3 pm and 300 pm the mass median diameter (MMD) was

determined to be

16.8 pm.

The particle size distribution was found to range from approximately 0.3 I~m to 35 pm.

284
RCC Study Number 845429 REPORT Page 14

TKA 40278 (CG1827)

Table 1 : Particle size distribution of TKA 40278 (CGI 827)

1. Run 2. Run 3. Run mean Stand.

Particle Particle ~article Particle Dev.

Size Size Size Size

% pm pm IJm pm [±]

5 < 1.82 < 1.80 < 2.01 < 1.9 0.116

10 < 4.95 <5.11 < 4.73 < 4.9 0.191

50 < 16.80 < 16.90 < 16.82 < 16.8" 0.053

9O < 25.21 < 25.08 < 27.53 < 25.9 1.379

*) The MMD of the test item indicates that 50 % of the particles are smaller than 16.8 pm.

285
RCC Study Number 845429 REPORT Page 18

TKA 40278 (CGI 827)

Table 2: Particle size distribution of TKA 40278 (CGI 827) in % at the relevant particle size

1. Run 2. Run 3. Run Mean Stand.

pm Dev.

% % % %

< 0.3

< 0.5 0,37 0.45 0.29 0.37 0.08

< 1.0 2.29 2.49 2.27 0.24

< 2.0 5.47 5.47 4.98 5.31 0.28

< 3.0 7.44 7.34 6.99 7.26 0.24

< 4.0 8.85 8.72 8.74 8.77 0.07

< 5.0 10.06 9.88 10.46 10.13 0.30

< 6.0 11.21 10.92 12.20 11.44 0.67

< 7.0 12.40 11.96 14.05 12.80 1.10

< 8.0 13.77 13,16 16.10 14.34 1.55

< 9.0 15.48 14.69 18.46 16.21 1.99

< 10.0 17.74 16.77 21.24 18.58 2.35

< 11.0 20.68 19.58 24.47 21.58 2.57

< 12.0 24.33 23.17 28.12 25.21 2.59

< 13.0 28,61 27.43 32.13 29.39 2.45

< 14.0 33.64 32.54 36.53 34.24 2.06

< 15.0 39.24 38.27 41.19 39.57 1.49

< 16.0 45.07 44.26 45.97 45.10 0.86

< 17.0 51.22 50.61 50.87 50.90 0.31

< 18.0 57.31 56,90 55.73 56.65 0.82

< 19,0 63.15 62.93 60.44 62.17 1.51

< 20.0 68.92 68.87 65.11 67.63 2.19

< 22.0 78.73 78.91 73.49 77.04 3.08

< 24.0 86.18 86.45 80.49 84.37 3.37

< 26.0 91.71 91.98 86,22 89.97 3,25

< 28.0 95,80 96.02 90.96 94.26 2.86

< 30.0 97.93 98.08 94.11 96.71 2.25

< 35.0 100.0o 100.00 98.95 99.65 0.61

< 40.0 100.00 100.00 100.00 100.00 0.00

< 45.0 100.00 100,00 100.00 100.00 0.00

< 50.0 100.00 100.00 100.00 I00.00 0.00

286
RCC Study Number 845429 REPORT Page 16

TKA 40278 (CGI 827)

Figure 1: Particle size distribution curve of TKA 40278 (CGI 827) (1. Run)

FRITSCH PARTICLE SIZER A N A L Y S E T T E 2 ?

RCC ~d,

13 I 01-24.2003 112:181

RCC Study No 845429

’TKA 40278 (CG1827)

’In Petroleumbenzin (60-80°C)

’RCC No 128115 / Batch KL 01/02

’l.Zugabe. 1. Msssung

~4.01.2003

I UserlD LAB/22 ISarN°" SN3165 I

Me~3berelch 0.31 [,um] - 300.74 [pm] Pumpe 100 [rpm] Aufl5sung 62 MeSkan&le ( 17 mm/ 114 mm ) ROhrer 2 [rpm] Absorption 7.00 [%] Ultraschall 0

Dauerder Messung 12 [Scans]

Mo<:le]l Independant

Fraunhofer Berechnung aktiv.

Inte rpolationswerte... C:XA22WlN~RITSC Hk03_50.FPS

****** % < 0.80 ,urn 0.37 % < 0.50 ,urn 2.29 % < 1,00 ,urn 5.47 % < 2.00 pm 7.44 % < 3.00 pm 8.85 % < 4,00 .urn 10,06%< 5.00 pm 11~21%< 6.00 #m 12.40%< 7,00 ,urn 13.77%< 8.00 pm 15.48%,< 9.00 pm 17,74%< 10.00 pm 20.68%< 11,00 ,urn 24,33%< 12,00 pm 28,61%< 18.00 pm 33,64%< 14.00 .urn 89.24%< 15.00 ,urn 45.07%< 16.00 pm 81.22%< 17.00 pm 57,31%< 18.00 pm 63.15%< 19,00 pm 68,92%< 20,00 pm 78.73%< 22.00 pm " 86,18%< 24,00 ,urn 91,71%< 26,00 .urn 95.80%< 28,00 pm 97,93%< 30.00 pm 100.00’/o < 35,00 pm 100.00% < 40.00 pm 100.00% < 45.00 ,urn 100.00’~ < 50,00 pm

Interpolationswerte_. C:~,22WIN~RITSCH~5_90PERC.FPV

5.00%< 1.82 pm 10,00%< 4,95 ,urn 50,00%< 16.80 .um 90,00% < 25.21 pm

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