International Federation Determination of Ethanol IFU Analysis

of Fruit Juice Producers by No. 2 (Rev. 2005)

(IFU) Gas Chromatography Page 1 of 3

Principle

The ethanol content of a fruit juice is determined by gas chromatography using a

packed column. This analysis will also allow the determination of the concentrations
of methanol and acetaldehyde in a beverage/juice. The quantification is carried out
using the internal standard approach. This type of analysis can also be carried out
using a capillary column.

WORKING INSTRUCTIONS

1. Apparatus and reagents

Gas chromatograph with FID.

Packed separation column: 2 m glass, i.d. 2 mm, filled with 6.6 % Carbowax 20M on
Carbopack B.
All reagents and standards should be of analytical grade
Citric acid
Ethyl methyl ketone (internal standard)
Ethanol
Methanol
Acetaldehyde

Mixed standard
Prepare a mixed standard solution on a volume basis as described below: -
0.1 % vol. ethanol ( approx. ≡ 789 mg/l)
0.02 % vol. methanol (approx. ≡ 158 mg/l)
0.01 % vol. acetaldehyde (approx. ≡ 78,3 mg/l) (cool to -20°C before addition, to limit
evaporation)
0.1% vol. ethyl methyl ketone (approx. ≡ 805 mg/l) as an intemal standard

Note: - The acetaldehyde should be cooled to –20°C to ensure that none of the
standard evaporates during its addition. Or prepare it on a weight basis by adding the
acetaldehyde to the volumetric flask containing the other standards and some water.

2. Determination procedure

A) Preparation of samples

To 10.0 ml of fruit juice (filtered if necessary) add 10 µl of ethyl methyl ketone, 0.2 g
of citric acid and mix well (sample solution).
International Federation Determination of Ethanol IFU Analysis
of Fruit Juice Producers by No. 2 (Rev. 2005)
(IFU) Gas Chromatography Page 2 of 3

B Gas chromatography conditions

Packed column as above

Carrier gas He or N2 with a head pressure of 200 kPa.
Column temperature: 90 oC isothermal
Injector and detector temperature 200 oC
Injection volume: 2 µl

3. Calculation and statement of results

The concentration of the ethanol, or other materials, is determined by reference to

the internal standard either using peak heights or areas. Values should be quoted in
mg/l.

4. Remarks

For ethanol values above 10 mg/l a direct injection can be used as this gives
reproducible results. For ethanol levels below 10 mg/l the sample should be
subjected to steam distillation prior to analysis by GC. It is recommended to pre-distill
samples if they are being analysed by capillary GC, this will prolong the working life
of the column.

Both free and total methanol can be measured using this method. If the total
methanol level is required refer to reference 1 for details of how to determine.

Note: - Although no direct references could be found for the analysis of ethanol in
fruit juices using capillary GC, reference 2 is a method that should be adaptable for
this matrix, but is actually a reference for wines and spirits. Reference 3 is a
collaborative trial for the determination of ethanol in fruit juices by GC, including
details for one collaborator using a Capillary GC approach.

5. Literature

1) H. Tanner and H. Limacher, Schweiz. Z. Obst- und Weinbau 120, 163-166

(1984)
2) Anon. Schweizerische Lebensmittelbuch, Ausgabe 2001. Published by the
Swiss Federal Office of Public Health Method No. 32/6
3) Anon. Draft BS:pEN13690:1999 available from British Standards in UK
International Federation Determination of Ethanol IFU Analysis
of Fruit Juice Producers by No. 2 (Rev. 2005)
(IFU) Gas Chromatography Page 3 of 3

Figures: Example chromatograms using packed column as described above.

Figure 1: Standard solution (2.0 µl) Figure 2: Orange juice (2.0 µl)

1. Acetaldehyde; 2. Methanol; 3. Acetone; 1. Acetaldehyde; 4. Ethanol; 5. Ethyl methyl

4. Ethanol; 5. Ethyl methyl ketone; 6. Ethyl ketone; 6. Ethyl acetate; 8. Water;
acetate; 7. Propan-1-ol; 8. Water 9.-11. Unknown pyrolysis products (may be
suppressed by addition of citric acid)