Commission Regulation (EC) No 128/2004 of 23 January 2004 amending Regulation (EEC) No 2676/90 determining Community methods for the analysis of wines
Date: 2004-01-23
Institutions: European Commission
Subjects: alcohol, analytical chemistry, fruit product, product quality, wine

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COMMISSION REGULATION (EC) No 128/2004

of 23 January 2004

amending Regulation (EEC) No 2676/90 determining Community methods for the analysis of
wines

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

Having regard to the Treaty establishing the European
Community,

Having regard to Council Regulation (EC) No 1493/1999 of 17
May 1999 on the common organisation of the market
in
wine (1), and in particular Article 46(3) thereof,

Whereas:

(1)

(2)

(3)

The method for measuring the alcoholic strength of
wine by hydrostatic balance has been updated and vali-
dated in accordance with internationally recognised
criteria. The International Vine and Wine Office adopted
the new description of
its General
Assembly in 2003.

this method at

this measurement method can constitute a
Use of
simpler and more accurate means of checking the alco-
holic strength by volume of wines and thereby avoid the
disputes arising from the use of less accurate methods.

The updated description of this method, together with
the experimental values for the validation parameters of
the method, should be included in Chapter 3 of the
Annex to Commission Regulation (EEC) No 2676/90 (2).

(4)

(5)

Regulation (EEC) No 2676/90 should therefore be
amended accordingly.

The measures provided for in this Regulation are in
accordance with the opinion of
the Management
Committee for Wine,

HAS ADOPTED THIS REGULATION:

Article 1

Chapter 3 of the Annex to Regulation (EEC) No 2676/90 ‘Alco-
holic strength by volume’ is hereby amended as follows:

1.

in paragraph 2, point 2.3.2 is deleted;

2. the text in the Annex to this Regulation is inserted as para-

graph 4a after paragraph 4,

3.

in paragraph 5, point 5.2 ‘Densimetry using a hydrostatic
balance’ is deleted.

Article 2

This Regulation shall enter into force on the seventh day
following its publication in the Official Journal of the European
Union.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 23 January 2004.

For the Commission

Franz FISCHLER

Member of the Commission

(1) OJ L 179, 14.7.1999, p. 1. Regulation as last amended by Regu-

lation (EC) No 1795/2003 (OJ L 262, 14.10.2003, p. 13).

(2) OJ L 272, 3.10.1990, p. 1. Regulation as last amended by Regu-

lation (EC) No 440/2003 (OJ L 66, 11.3.2003, p. 15).

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‘4a.

VALIDATED METHOD

ANNEX

Determination of the alcoholic strength of wines using a hydrostatic balance

1.

METHOD OF MEASUREMENT

1.1.

Introduction

Alcoholic strength by volume must be measured before marketing, principally to comply with labelling
rules.

Alcoholic strength by volume is equal to the number of litres of ethanol contained in 100 litres of
wine measured at 20 °C. Referred to as “ % vol.”.

1.2.

Object and field of application

The method of measurement described is densimetry using a hydrostatic balance.

For the purposes of the regulatory provisions in force, the trial temperature is set at 20 °C.

1.3.

Principle and definitions

The principle of this method is based on distilling wine volume by volume. The distilling method is
described in this Chapter. Distillation eliminates non-volatile substances. Homologues of ethanol,
together with the ethanol and ethanol homologues in ethyl esters, are included in the alcoholic
strength since they occur in the distillate.

The density of the distillate obtained is then measured. The density of a liquid at a given temperature
is equal to the quotient of the mass over its volume: ρ

2 = m/V; for wine, it is expressed in g/ml.

The alcoholic strength of wines can be measured by densimetry using a hydrostatic balance based on
Archimedes' principle, according to which a body immersed in a liquid receives a vertical upward
thrust from the liquid equal to the weight of liquid displaced.

1.4.

Reagents

During the analysis, unless otherwise is stated, use only reagents of recognised analytical grade and
water of at least grade 3 as defined in ISO 3696:1987.

1.4.1.

Float-cleaning solution (sodium hydroxide, 30 % w/v)

To prepare 100 ml, weigh 30 g sodium hydroxide and make up to volume using 96 % volume
ethanol.

1.5.

Apparatus and equipment

Usual laboratory apparatus and in particular the following:

1.5.1.

Single-pan hydrostatic balance with a sensitivity of 1 mg.

1.5.2.

Float with a volume of at least 20 ml, specially adapted to the balance, suspended with a thread of
diameter not exceeding 0,1 mm.

1.5.3. Measuring cylinder bearing a level mark. The float must be capable of being contained completely
within the volume of the cylinder located below the mark; the surface of the liquid may be penetrated
only by the supporting thread. The measuring cylinder must have an internal diameter at least 6 mm
larger than that of the float.

1.5.4.

Thermometer (or temperature-measuring probe) graduated in degrees and tenths of a degree from
10 °C to 40 °C, calibrated to 0,05 °C.

1.5.5. Weights, calibrated by a recognised certifying body.

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1.6.

Procedure

The float and measuring cylinder must be cleaned between each measurement with distilled water,
dried with soft laboratory paper which does not shed fibres and rinsed with the solution whose density
is to be determined. Measurements must be made as soon as the apparatus has reached stability so as
to restrict alcohol loss by evaporation.

1.6.1.

Calibration of the balance

Although balances usually have an internal calibration system, the hydrostatic balance must be capable
of calibration with weights checked by an official certifying body.

1.6.2.

Calibration of the float

1.6.2.1. Fill the measuring cylinder to the mark with double-distilled water (or water of equivalent purity, e.g.
microfiltered water with a conductivity of 18,2 MΩ/cm) at a temperature between 15 °C and 25 °C
but preferably at 20 °C.

1.6.2.2. Immerse the float and the thermometer, stir, read off the density of the liquid from the apparatus and,

if necessary, correct the reading so that it is equal to that of the water at measurement temperature.

1.6.3.

Control using a water-alcohol solution

1.6.3.1. Fill the measuring cylinder to the mark with a water-alcohol mixture of known strength at a tempera-

ture between 15 °C and 25 °C but preferably at 20 °C.

1.6.3.2. Immerse the float and the thermometer, stir, read off the density of the liquid (or the alcoholic strength
if this is possible) from the apparatus. The alcoholic strength thus established should be equal to the
previously determined alcoholic strength.

Note: This solution of known alcoholic strength can also be used to calibrate the float instead of

double-distilled water.

1.6.4. Measurement of the density of a distillate (or of its alcoholic strength if the apparatus allows)

1.6.4.1. Pour the test sample into the measuring cylinder up to the graduation mark.

1.6.4.2. Immerse the float and the thermometer, stir, read off the density of the liquid (or the alcoholic strength
if this is possible) from the apparatus. Note the temperature if the density is measured at t °C ρ

t.

1.6.4.3. Correct ρ
Chapter).

t using the table of densities ρ

t for water-alcohol mixtures (Table II of Annex II to this

1.6.5.

Cleaning of float and measuring cylinder

1.6.5.1. Immerse the float in the float-cleaning solution in the measuring cylinder.

1.6.5.2. Allow to soak for one hour spinning the float periodically.

1.6.5.3. Rinse with copious amounts of tap water followed by distilled water.

1.6.5.4. Dry with soft laboratory paper which does not shed fibres.

Carry out this procedure when the float is first used and then regularly as required.

1.6.6.

Result

Using the density ρ
20, calculate the real alcoholic strength using the table giving the value of the alco-
holic strength by volume ( % vol.) at 20 °C as a function of the density at 20 °C of water-alcohol
mixtures, i.e. the international table adopted by the International Legal Metrology Organisation in its
Recommendation No 22.

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2.

Official Journal of the European Union

27.1.2004

COMPARISON OF MEASUREMENTS MADE USING A HYDROSTATIC BALANCE WITH PROCE-
DURES OBTAINED USING AN ELECTRONIC DENSIMETER

Using samples with an alcoholic strength between 4 % and 18 % vol., repeatability and reproducibility
were measured following an inter-laboratory ring test. The alcoholic strength of different samples as
measured using the hydrostatic balance and the electronic densimeter and the repeatability and repro-
ducibility values derived from an extensive multiannual inter-comparison exercise were compared.

2.1.

Samples

Wines of different density and alcoholic strength prepared each month on industrial scale, taken from
a properly stored stock of bottles and delivered as anonymous products to the laboratories.

2.2.

Laboratories

Laboratories participating in the monthly ring test organised by the Unione Italiana Vini (Verona, Italy)
according to ISO 5725 (UNI 9225) rules and the International Protocol of Proficiency Testing for
chemical analysis laboratories established by AOAC, ISO and IUPAC and ISO 43 and ILAC G13 guide-
lines. An annual report is to be supplied by this company to all participants.

2.3.

Equipment

2.3.1.

Electronic hydrostatic balance (accurate to 5 decimal places), if possible with a data processing device:

2.3.2.

Electronic densimeter, if possible with autosampler.

2.4.

Analysis

According to the rules for the validation of methods, each sample was analysed twice consecutively to
determine the alcoholic strength.

2.5.

Result

Table 1 shows the results of the measurements obtained by the laboratories using the hydrostatic
balance.

Table 2 shows the results obtained by the laboratories using an electronic densimeter.

2.6.

Evaluations of the results

2.6.1.

The trial results were examined for evidence of individual systematic error (p < 0,025) using Cochran's
and Grubb's tests successively, by procedures described in the internationally agreed Protocol for the
Design, Conduct and Interpretation of Method-Performance Studies.

2.6.2.

Repeatability (r) and reproducibility (R)

Calculations for repeatability (r) and reproducibility (R) as defined by that protocol were carried out on
those results remaining after the removal of outliers. When assessing a new method there is often no
validated reference or statutory method with which to compare precision criteria, hence it is useful to
compare the precision data obtained from a collaborative trial with “predicted” levels of precision.
These “predicted” levels are calculated from the Horwitz equation. Comparison of the trial results and
the predicted levels give an indication as to whether the method is sufficiently precise for the level of
analyte being measured. The Horwitz predicted value is calculated from the Horwitz equation.

RSDR = 2(1-0,5 logC)

where C = measured concentration of analyte expressed as a decimal (e.g. 1 g/100 g = 0,01).

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The Horrat value gives a comparison of the actual precision measured with the precision predicted by
the Horwitz equation for a method measuring at that particular level of analyte. It is calculated as
follows:

HoR = RSDR(measured)/RSDR(Horwitz )

2.6.3.

Interlaboratory precision

A Horrat value of 1 usually indicates satisfactory inter-laboratory precision, whereas a value of 2
usually indicates unsatisfactory precision, i.e. one that is too variable for most analytical purposes or
where the variation obtained is greater than that expected for the type of method employed. Hor is
also calculated, and used to assess intra-laboratory precision, using the following approximation:

RSDr(Horwitz) = 0,66 RSDR(Horwitz) (this assumes the approximation r = 0,66 R).

Table 3 shows the differences between the measurements obtained by laboratories using electronic
densimetry and those using a hydrostatic balance. Excluding sample 2000/3, which had very low
alcohol strength and for which both techniques show poor reproducibility, a very good agreement is
generally observed for all other samples.

2.6.4.

Fidelity parameters

Table 4 shows the average overall fidelity parameters computed from all monthly trials carried out
from January 1999 until May 2001.

In particular:

Repeatability (r) = 0,074 ( % vol.)
densimetry;

for the hydrostatic balance and 0,061 ( % vol.)

for electronic

Reproducibility (R) = 0,229 ( % vol.) for the hydrostatic balance and 0,174 ( % vol.) for electronic
densimetry.

2.7.

Conclusion

The results of determining the alcoholic strength of a wide range of wines show the comparability of
measurements carried out with the hydrostatic balance and the electronic density-meter using a
frequency oscillator and demonstrate that the values of the validation parameters are similar for both
methods.

— Mean

— N

— Nc

— Outliers

— n1

— R

— Sr

— RSDr

— Hor

— R

— SR

— HoR

Ke y to table s

the mean of all the data used in the statistical analysis

total number of sets of data submitted

number of results excluded from statistical analysis due to non-compliance

number of results excluded from statistical analysis due to determination as outliers
by either Cochran's or Grubb's tests

number of results used in the statistical analysis

repeatability limit

the standard deviation of the repeatability

the relative standard deviation of the repeatability (Sr × 100/Mean)

the Horrat value for repeatability is the observed RSDr divided by the RSDr value
estimated from the Horwitz equation using the assumption r = 0,66R

reproducibility limit

the standard deviation of the reproducibility

the Horrat value for reproducibility is the observed RSDR value divided by the
RSDR value calculated from HoR = RSDR(measured)/RSDR

Mean

1999/1

11,043

1999/2

11,247

1999/3

11,946

1999/4

7,653

1999/5

11,188

1999/6

11,276

1999/7

8,018

1999/9

11,226

1999/10

11,026

1999/11

7,701

1999/12

10,987

2000/1

11,313

2000/2

11,232

2000/3

0,679

2000/4

11,223

2000/5

7,439

2000/6

11,181

2000/7

10,858

2000/9

12,031

2000/10

11,374

2000/11

7,644

2000/12

11,314

2001/1

11,415

2001/2

11,347

2001/3

11,818

2001/4

11,331

2001/5

8,063

n

17

14

16

17

17

19

17

17

17

16

17

16

17

10

18

19

19

16

17

18

18

19

19

19

16

17

19

Outliers

1

1

0

1

0

0

0

0

0

1

2

0

0

0

0

1

0

0

1

0

0

1

0

0

0

0

1

n1

16

13

16

16

17

19

17

17

17

15

15

16

17

10

18

18

19

16

16

18

18

18

19

19

16

17

18

Table 1: Hydrostatic balance (HB)

r

Sr

RSDr

Hor

R

sR

RSDR

HoR

No of
replicates

Critical
difference
CrD95

0,0571

0,0204

0,1846

0,1004

0,1579

0,0564

0,0584

0,0208

0,1854

0,1011

0,1803

0,0644

0,0405

0,0145

0,1211

0,0666

0,1593

0,0569

0,0502

0,0179

0,2344

0,1206

0,1537

0,0549

0,0871

0,0311

0,2780

0,1515

0,2701

0,0965

0,0846

0,0302

0,2680

0,1462

0,2957

0,1056

0,0890

0,0318

0,3964

0,2054

0,2573

0,0919

0,0580

0,0207

0,1846

0,1423

0,2796

0,0999

0,0606

0,0216

0,1961

0,1066

0,2651

0,0947

0,0643

0,0229

0,2980

0,1535

0,2330

0,0832

0,0655

0,0234

0,2128

0,1156

0,1258

0,0449

0,0986

0,0352

0,3113

0,1699

0,2577

0,0920

0,0859

0,0307

0,2731

0,1489

0,2535

0,0905

0,5107

0,5727

0,4764

0,7172

0,8622

0,9365

1,1462

0,8896

0,8588

1,0805

0,4089

0,8135

0,8060

0,0680

0,0243

3,5773

1,2783

0,6529

0,2332

34,3395

0,0709

0,0253

0,2257

0,1230

0,2184

0,0780

0,0630

0,0225

0,3023

0,1549

0,1522

0,0544

0,0536

0,0191

0,1710

0,0932

0,2783

0,0994

0,0526

0,0188

0,1731

0,0939

0,1827

0,0653

0,0602

0,0215

0,1787

0,0985

0,2447

0,0874

0,0814

0,0291

0,2555

0,1395

0,2701

0,0965

0,0827

0,0295

0,3863

0,1988

0,2289

0,0817

0,0775

0,0277

0,2447

0,1336

0,2421

0,0864

0,0950

0,0339

0,2971

0,1623

0,2410

0,0861

0,0792

0,0283

0,2493

0,1361

0,1944

0,0694

0,0659

0,0235

0,1990

0,1093

0,2636

0,0941

0,1067

0,0381

0,3364

0,1836

0,1895

0,0677

0,0782

0,0279

0,3465

0,1797

0,1906

0,0681

0,6951

0,7307

0,8890

0,6011

0,7263

0,8482

1,0694

0,7641

0,7539

0,6119

0,7965

0,5971

0,8442

0,18

0,21

0,17

0,24

0,31

0,34

0,39

0,45

0,31

0,37

0,15

0,29

0,29

8,10

0,25

0,25

0,32

0,22

0,26

0,31

0,36

0,28

0,27

0,22

0,29

0,22

0,29

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

0,1080

0,1241

0,1108

0,1057

0,1860

0,2047

0,1764

0,1956

0,1850

0,1616

0,0827

0,1754

0,1740

0,4604

0,1503

0,1029

0,1950

0,1265

0,1704

0,1866

0,1565

0,1667

0,1636

0,1316

0,1834

0,1229

0,1290

L

1
9
/
8

E
N

O

f
f
i
c
i
a
l

J
o
u
r
n
a
l

o
f

t
h
e

E
u
r
o
p
e
a
n
U
n
o
n

i

.

.

2
7
1
2
0
0
4

Mean n1

D1999/1

11,019

D1999/2

11,245

D1999/3

11,967

D1999/4

7,643

D1999/5

11,188

D1999/6

11,303

D1999/7

8,026

D1999/9

11,225

D1999/10

11,011

D1999/11

7,648

D1999/12

10,999

D2000/1

11,248

D2000/2

11,240

D2000/3

0,526

D2000/4

11,225

D2000/5

7,423

D2000/6

11,175

D2000/7

10,845

D2000/9

11,983

D2000/10

11,356

D2000/11

7,601

D2000/12

11,322

D2001/1

11,427

D2001/2

11,320

D2001/3

11,826

D2001/4

11,339

D2001/5

8,058

n

18

19

21

19

21

21

21

17

19

21

16

22

19

12

19

21

23

21

22

22

27

25

29

29

34

31

28

Outliers

1

2

0

1

3

0

0

0

0

1

1

1

3

1

1

0

2

5

1

1

0

1

0

1

1

2

0

n1

17

17

21

18

18

21

21

17

19

20

15

21

16

11

18

21

21

16

21

21

27

24

29

28

33

29

28

Table 2: Electronic densimetry (ED)

r

sr

RSDr

Hor

R

sR

RSDR

HoR

No of
replicates

Critical
difference
CrD95

.

.

2
7
1
2
0
0
4

0,0677

0,0242

0,2196

0,1193

0,1996

0,0713

0,0448

0,0160

0,1423

0,0776

0,1311

0,0468

0,0701

0,0250

0,2091

0,1151

0,1552

0,0554

0,0610

0,0218

0,2852

0,1467

0,1340

0,0479

0,0260

0,0093

0,0829

0,0452

0,2047

0,0731

0,0652

0,0233

0,2061

0,1125

0,1466

0,0523

0,0884

0,0316

0,3935

0,2039

0,1708

0,0610

0,0372

0,0133

0,1183

0,0645

0,1686

0,0602

0,0915

0,0327

0,2969

0,1613

0,1723

0,0615

0,0615

0,0220

0,2872

0,1478

0,1538

0,0549

0,0428

0,0153

0,1389

0,0755

0,2015

0,0720

0,0697

0,0249

0,2212

0,1206

0,1422

0,0508

0,0448

0,0160

0,1424

0,0776

0,1619

0,0578

0,6470

0,4165

0,4631

0,6262

0,6536

0,4631

0,7600

0,5366

0,5588

0,7183

0,6541

0,4516

0,5145

0,23

0,15

0,17

0,21

0,24

0,17

0,26

0,19

0,20

0,24

0,23

0,16

0,19

0,0327

0,0117

2,2185

0,7630

0,9344

0,3337

63,4009

14,39

0,0476

0,0170

0,1514

0,0825

0,1350

0,0482

0,0628

0,0224

0,3019

0,1547

0,2635

0,0941

0,0606

0,0217

0,1938

0,1056

0,1697

0,0606

0,0440

0,0157

0,1449

0,0786

0,1447

0,0517

0,0841

0,0300

0,2507

0,1380

0,2410

0,0861

0,0635

0,0227

0,1997

0,1090

0,1865

0,0666

0,0521

0,0186

0,2448

0,1258

0,1685

0,0602

0,0476

0,0170

0,1503

0,0820

0,1594

0,0569

0,0706

0,0252

0,2207

0,1206

0,1526

0,0545

0,0675

0,0241

0,2128

0,1161

0,1570

0,0561

0,0489

0,0175

0,1476

0,0811

0,1762

0,0629

0,0639

0,0228

0,2012

0,1099

0,1520

0,0543

0,0473

0,0169

0,2098

0,1088

0,2025

0,0723

0,4295

1,2677

0,5424

0,4766

0,7183

0,5866

0,7916

0,5028

0,4771

0,4952

0,5322

0,4788

0,8976

0,15

0,43

0,20

0,17

0,26

0,21

0,27

0,18

0,17

0,18

0,19

0,17

0,31

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

0,1370

0,0900

0,1040

0,0897

0,1442

0,0984

0,1124

0,1178

0,1129

0,1043

0,1408

0,0944

0,1123

0,6605

0,0924

0,1836

0,1161

0,0999

0,1651

0,1280

0,1162

0,1102

0,1020

0,1057

0,1222

0,1026

0,1412

E
N

O

f
f
i
c
i
a
l

J
o
u
r
n
a
l

o
f

t
h
e

E
u
r
o
p
e
a
n
U
n
o
n

i

L

1
9
/
9

n

17

14

16

17

17

19

17

17

17

16

17

16

17

10

18

19

19

16

17

18

18

19

19

19

16

17

19

1999/1

1999/2

1999/3

1999/4

1999/5

1999/6

1999/7

1999/9

1999/10

1999/11

1999/12

2000/1

2000/2

2000/3

2000/4

2000/5

2000/6

2000/7

2000/9

2000/10

2000/11

2000/12

2001/1

2001/2

2001/3

2001/4

2001/5

Mean (HB)

11,043

11,247

11,946

7,653

11,188

11,276

8,018

11,226

11,026

7,701

10,987

11,313

11,232

0,679

11,223

7,439

11,181

10,858

12,031

11,374

7,644

11,314

11,415

11,347

11,818

11,331

8,063

(*) Test 2000/3 is not taken into account.

Table 3: Comparison of results between hydrostatic balance (HB) and electronic densimetry (DE)

Outliers

1

1

0

1

0

0

0

0

0

1

2

0

0

0

0

1

0

0

1

0

0

1

0

0

0

0

1

n1

16

13

16

16

17

19

17

17

17

15

15

16

17

10

18

18

19

16

16

18

18

18

19

19

16

17

18

D1999/1

D1999/2

D1999/3

D1999/4

D1999/5

D1999/6

D1999/7

D1999/9

D1999/10

D1999/11

D1999/12

D2000/1

D2000/2

D2000/3

D2000/4

D2000/5

D2000/6

D2000/7

D2000/9

D2000/10

D2000/11

D2000/12

D2001/1

D2001/2

D2001/3

D2001/4

D2001/5

Mean (ED)

11,019

11,245

11,967

7,643

11,188

11,303

8,026

11,225

11,011

7,648

10,999

11,248

11,240

0,526

11,225

7,423

11,175

10,845

11,983

11,356

7,601

11,322

11,427

11,320

11,826

11,339

8,058

n

18

19

21

19

21

21

21

17

19

21

16

22

19

12

19

21

23

21

22

22

27

25

29

29

34

31

28

Outliers

1

2

0

1

3

0

0

0

0

1

1

1

3

1

1

0

2

5

1

1

0

1

0

1

1

2

0

n1

17

17

21

18

18

21

21

17

19

20

15

21

16

11 (*)

18

21

21

16

21

21

27

24

29

28

33

29

28

Average difference/D TAV (HB-ED)

Standard deviation on difference

∆TAV(HB-ED)

0,024

0,002

– 0,021

0,010

0,000

– 0,028

– 0,008

0,002

0,015

0,052

– 0,013

0,065

– 0,008

0,153

– 0,002

0,016

0,006

0,013

0,049

0,018

0,043

– 0,008

– 0,012

0,027

– 0,008

– 0,008

0,004

0,014

0,036

L

1
9
/
1
0

E
N

O

f
f
i
c
i
a
l

J
o
u
r
n
a
l

o
f

t
h
e

E
u
r
o
p
e
a
n
U
n
o
n

i

.

.

2
7
1
2
0
0
4

27.1.2004

EN

Official Journal of the European Union

L 19/11

n1

Weighted repeatability variance

R

Sr

Weighted reproducibility variance

R

sR

Table 4: Fidelity parameters

Hydrostatic balance

Electronic densimetry

441

557

0,309

0,074

0,026

2,948

0,229

0,082

0,267

0,061

0,022

2,150

0,174

0,062’