Study Title 

Study Director 

Test Facility 

Sponsor 

Internal distribution 

Study No. 

FINAL REPORT 

Determination of the density of 

according to OECD Guideline No. 109 (pycnometer) 

Archives 

Author. 
Study No. - Final report 

CONTENTS: 

Title page 

Contents 

Statement of GLP compliance (Swiss GLP monitoring authorities) 

Statement of GLP compliance (Study Director) and signatures 

Quality Assurance Unit 

Summary 

1 . General information 

2. Information on the test substance 

2. 1 . Identity 

3. 

4. 

2.2. Physical-chemical and analytical data 

General test method 

Results 

Total number of pages : 12 

Page 2/12 

Page 

1 

2 

3 

4 

5 

6 

7 

8 

8 

8 

9-11 

12 
Study No. - Final report Page 3/12 

The Swiss GLP Monitoring Authorities 

Swiss Federal 

Office of 

Public Health 

It is hereby confirmed that 

during the period of 

the following Test Facility of 

Swiss Agency for the 

Environment, Forests 

and Landscape 

March 31 - April 1, 2005 

I 

lntercantonal Office 

for the Control of 

Medicines 

was inspected by the Swiss Agency for the Environment, Forests and Landscape with 

respect to the compliance with the Swiss legislation on Good Laboratory Practice. 

Test Facility Areas of expertise 

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

Inspections and Audits. It was found that the aforementioned test facility was operating in 

compliance with the Swiss Ordinance on Good Laboratory Practice [SR 813.016.5] at the 

time it was inspected. 

Bern, June 2005 

Swiss Agency for the Environment, 

Forests and Landscape 

Philippe Roch 
Study No. - Final report Page 4/12 

STATEMENT OF COMPLIANCE WITH GOOD LABORATORY PRACTICE: 

Study No. 

Test substance 

Title Determination of the density of according to 

OECD Guideline No. 109 (pycnometer). 

This study has been performed in compliance with the Swiss Ordinance relating to 

Good Laboratory Practice, adopted May 18th, 2005 [RS 813.112.1 ]. 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 undersigned declare that this study was carried out according to the procedures 

herein described and this report represents a true and accurate record of the results 

obtained. 

Study director 

Test Facility Management: 

Date: 
Study No. - Final report Page 5/12 

QUALITY ASSURANCE UNIT 

STATEMENT 

Study No. 

Test substance 

Study title Determination of the density of 

according to OECD Guideline No. 109 (pycnometer). 

Test facilities are periodically inspected and the results are reported to the responsible 

persons and the management. 

Experimental procedures were periodically inspected. The study plan and this final 

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 

February 1 8

\ 2007 

February 1st, 2007 

Study plan February 1st, 2007 

Weighting and filling of the February 1

8

\ 2007 

pycnometer with the displacement 

liquid 

Final report 

The methods and procedures as used in the reference study are accurately described 

in this document and reported results reflect accurately the raw data of the study. 

Manager, Quality Assurance Unit 

Date 
Study No. - Final report Page 6/12 

SUMMARY 

The density of was determined according to the OECD guideline No. 1 09 

(pycnometer). The result can be expressed as follows: 

p = 1 .12 g/cm3 (at 25°C) 
Study No. - Final report Page 7/12 

1. GENERAL INFORMATION 

Study No. 

Purpose 

Guidelines 

Study Director 

Laboratory 

Technician 

Schedule: 

Study initiation date 

Start of experiments : 

End of experiments : 

Study completion date: 

Archives 

Determining the density of using a pycnometer 

OECD Guideline for the Testing of Chemicals No. 109, Paris, 

1995 

92/69/EEC, method A.3. 

January 2nd, 2007 

February 1 5

\ 2007 

April 3rd, 2007 

The study plan, all data generated and recorded during the 

study and the final report will be stored in the Archives of the 

for at 

least ten years after issue of the final report. 
Study No. - Final report Page 8/12 

2. INFORMATION ON THE TEST SUBSTANCE 

2.1 . Identity 

Commercial name 

Givaudan Code No. 

Chemical nature 

Empirical formula 

Molecular Weight 

CAS No. 

N/A 

2.2. Physical-chemical and analytical data 

-General 

Aspect 

- Sample tested 

Lot No. 

Purity (GC) 

Expiration date May 14,2007 
Study No. - Final report Page 9/12 

3. GENERAL TEST METHOD 

3.1 Introduction 

Many methods exist which allow to measure the density with a greater precision than is 

required for environmental assessment. 

The pycnometer method is simple and does not require any special or expensive 

equipment. 

The method requires the use of a "displacement liquid" in which the test substance is 

insoluble, which has good wetting properties for it and has a low vapour pressure. 

3.2 Definitions and units 

The density of a substance is the quotient of the mass m and its volume V: 

p= m/V 

Sl unit is kg/m3

, but g/cm3 is most often used. 

3.3 Principle 

The density of the solid substance is determined by measuring the difference of weight 

between a volume (the pycnometer) filled with the displacement liquid and the same 

volume filled with a known mass of the solid immersed in the displacement liquid. 

3.4 Equipment 

- a dessiccator whose lid can be equipped with a dropping funnel 

- a 25-ml pycnometer 

- a thermostated water bath (±0.1 °C) 

- a laboratory balance (± 0.1 mg) 

- a sieve with 500 flm mesh size 

3.5 Reference Substance 

No reference substance is required. 
Study No. - Final report Page 10/12 

3.6 Operating procedure 

3.6.1. Temperature 

The temperature of the determination has a marked effect on the density of the 

displacement liquid but not on that of the test substance. For more convenience, the 

temperature of the experiment (i.e. the temperature of the water-bath) was chosen 

about soc higher than room temperature (ca. 20°C). 

3.6.2 Displacement liquid 

is almost insoluble in water (less than 10 mg/1 at room temperature). A first 

test was made with pure water, but it completely failed since the test substance could 

not be correctly wetted with pure water. 

Thus a second test was made with a 1% Tween 80 aqueous solution. This was seen to 

be a good displacement liquid. 

3.6.3 Sample preparation 

The sample of test substance was thoroughly mixed and a specimen of about 60 g was 

sieved through a 500 !lm mesh sieve and dried in a dessiccator for one night. 

3.6.4 Density of the displacement liquid 

The pycnometer, empty, clean and dry, was weighed with its stopper: mass m1. It was 

then filled with deionised water and weighed: mass ms. 

After that it was filled with the displacement liquid and weighed: mass m4. 

The density of the displacement liquid was calculated as follows: 

with Po the density of water at the temperature of determination 

3.6.5 Density of the test substance 

The pycnometer, empty, clean and dry, was weighed with its stopper: mass m1 

The dry pycnometer was filled about to the half with test substance, stopped and 

weighed: mass m2 

The pycnometer was placed in the dessiccator and the dropping funnel affixed to the lid 

so that the stem penetrates into the pycnometer. The stopcock of the funnel was closed 

and vacuum was carefully applied to the dessiccator for 15 minutes after it has reached 

less than 2 kPa. 
Study No. - Final report Page 11/12 

The funnel was filled with displacement liquid and the stopcock slowly opened to allow 

the liquid to fill the pycnometer 15 mm above the level of the test substance. 

The stopcock was closed and vacuum applied again to the dessiccator (pressure less 

than 2 kPa) until any bubble in the liquid has disappeared. 

Air was allowed to enter the dessiccator, the pycnometer was removed, filled up with 

displacement liquid and placed in the water-bath for 30 minutes. 

The pycnometer was closed with its stopper, any excess of liquid was wiped off, and 

then the pycnometer was left 15 minutes on the balance scale and weighed: mass m3 . 

At the end of the experiment, the pycnometer was carefully rinsed and filled with 

deionised water, allowed to equilibrate in the water-bath for 30 minutes, closed and 

weighed after equilibration as already described in the previous paragraph: mass m4. 

The density of the test substance was calculated as follows: 

Ps = density of the test substance (g/cm3

) 

p1 = density of the displacement liquid (g/cm3

) 

m1 = weight (g) of the empty pycnometer with stopper 

m2 = weight (g) of the pycnometer containing test substance 

m3 weight (g) of the pycnometer filled with the test substance and the displacement 

liquid 

m4 weight (g) of the pycnometer filled with the displacement liquid 

The determination was made in duplicate. 
Study No. - Final report Page 12/12 

4. RESULTS 

Temperature of the determination: 25°C. The density of water at 25°C is 

(Logarithmische Rechnetafeln, Kuster, Thiel and Fischbeck, Walter de Gruyter and Co. 

ed., Berlin, 1965): 

p0 = 0.997071 g/cm3 

4. 1 First determination: 

m1 = 22.1758g m2 = 28.2578g m3 = 47.5520g m4 = 46.8906g m5 =46.8616g 

Density of the displacement liquid: 

p 1 = 0.998242 g/cm3 

Density of the test substance: 

Ps = 1.120 g/cm3 

4.2 Second determination 

m1 =24.165lg m2 =30.1875g m3 =46.9360g m4 =46.2803g m5 =46.2571g 

Density of the displacement liquid: 

p 1 = 0.998118g/cm3 

Density of the test substance: 

Ps 1.120 g/cm3 

4.3 Final result 

There is no difference between the two determinations. The average of the two 

determinations was then calculated and rounded off at two decimal places to express 

the desired density of 

Average: 

After rounding off: 

1.120 g/cm3 

1.12 g/cm3