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- Original (As adopted by EU)
Commission Regulation (EC) No 2870/2000 of 19 December 2000 laying down Community reference methods for the analysis of spirits drinks
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The method pertains to the determination of furfural, 5-hydroxymethylfurfural, 5-methylfurfural, vanillin, syringaldehyde, coniferaldehyde, sinapaldehyde, gallic acid, ellagic acid, vanillic acid, syringic acid and scopoletin, by high-performance liquid chromatography.
Analytical method recognised by the General Assembly of the International Organisation of Vine and Wine (OIV) and published by OIV under the reference OIV-MA-BS-16: R2009 .
Determination by high-performance liquid chromatography (HPLC), with detection by ultraviolet spectrophotometry at several wavelengths and by spectrofluorimetry.
The reagents must be of analytical quality. The water used must be distilled water or water of at least equivalent purity. It is preferable to use microfiltered water with a resistivity of 18,2 M Ω.cm.
Solvent A (0,5 % acetic acid) and solvent B (pure methanol). Filter through a membrane (porosity 0,45 μm). Degas in an ultrasonic bath, if necessary.
furfural: 5 mg/l; 5-hydroxymethyl furfural: 10 mg/l; 5-methylfurfural 2 mg/l; vanillin: 5 mg/l; syringaldehyde: 10 mg/l; coniferaldehyde: 5 mg/l; sinapaldehyde: 5 mg/l; gallic acid: 10 mg/l; ellagic acid: 10 mg/l; vanillic acid: 5 mg/l; syringic acid: 5 mg/l; scopoletin: 0,5 mg/l.
Standard laboratory apparatus
A spectrophotometric detector capable of measuring at wavelengths from 260 to 340 nm. It is however preferable to work with a multiple wavelength detector with a diode array or similar, in order to confirm the purity of the peaks.
A spectrofluorimetric detector — excitation wavelength: 354 nm, emission wavelength: 446 nm (for the trace determination of scopoletin; which is also detectable at 313 nm by spectrophotometry).
An injection device capable of introducing 10 or 20 μl (for example) of the test sample.
A high-performance liquid chromatography column, RP C18 type, 5 μm maximum particle size.
The reference solution and the spirit drink are filtered, if necessary, through a membrane with a maximum pore diameter of 0,45 μm.
Time: 0 min 50 min 70 min 90 min
solvent A (water-acid): 100 % 60 % 100 % 100 %
solvent B (methanol): 0 % 40 % 0 % 0 %
Note that in certain cases this gradient should be modified to avoid co-elutions.
Adapt the operating conditions so that the resolution factors of the peaks of all the compounds are equal to at least 1.
Express the concentration of each constituent in mg/l.
The following data were obtained in 2009 from an international method-performance study on a variety of spirit drinks, carried out following internationally-agreed procedures (1) (2).
Analyte | Furfural | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 15 | 15 | 15 | 15 | 15 | 15 |
No of results accepted (laboratories) | 14 | 12 | 13 | 14 | 13 | 13 |
Mean value (mg/l) | 2,9 | 1,2 | 1,7 | 10,6 | 15,3 | 13,9 |
Repeatability standard deviation, s r (mg/l) | 0,04 | 0,05 | 0,04 | 0,18 | 0,23 | 0,20 |
Repeatability relative standard deviation, RSD r (%) | 1,4 | 4,5 | 2,3 | 1,7 | 1,5 | 1,5 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,1 | 0,2 | 0,1 | 0,5 | 0,6 | 0,6 |
Reproducibility standard deviation, s R (mg/l) | 0,24 | 0,18 | 0,09 | 1,4 | 0,49 | 0,69 |
Reproducibility relative standard deviation, RSD R (%) | 8 | 15 | 5 | 13 | 3 | 5 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,7 | 0,5 | 0,3 | 3,8 | 1,4 | 1,9 |
Analyte | 5-Hydroxymethylfurfural | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 16 | 16 | 16 | 16 | 16 | 16 |
No of results accepted (laboratories) | 14 | 14 | 14 | 14 | 14 | 14 |
Mean value (mg/l) | 5,0 | 11,1 | 9,4 | 33,7 | 5,8 | 17,5 |
Repeatability standard deviation, s r (mg/l) | 0,09 | 0,09 | 0,09 | 0,42 | 0,07 | 0,13 |
Repeatability relative standard deviation, RSD r (%) | 1,7 | 0,8 | 1,0 | 1,3 | 1,2 | 0,8 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,2 | 0,3 | 0,3 | 1,2 | 0,2 | 0,4 |
Reproducibility standard deviation, s R (mg/l) | 0,39 | 1,01 | 0,50 | 4,5 | 0,4 | 1,6 |
Reproducibility relative standard deviation, RSD R (%) | 8 | 9 | 5 | 13 | 7 | 9 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 1,1 | 2,8 | 1,4 | 12,5 | 1,1 | 4,6 |
Analyte | 5-Methylfurfural | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 11 | 11 | 11 | 11 | 11 | 11 |
No of results accepted (laboratories) | 11 | 11 | 8 | 11 | 10 | 11 |
Mean value (mg/l) | 0,1 | 0,2 | 0,1 | 0,5 | 1,7 | 0,8 |
Repeatability standard deviation, s r (mg/l) | 0,01 | 0,01 | 0,02 | 0,02 | 0,03 | 0,07 |
Repeatability relative standard deviation, RSD r (%) | 10,7 | 6,1 | 13,6 | 4,7 | 2,0 | 10,0 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,0 | 0,0 | 0,1 | 0,1 | 0,1 | 0,2 |
Reproducibility standard deviation, s R (mg/l) | 0,03 | 0,04 | 0,03 | 0,18 | 0,20 | 0,26 |
Reproducibility relative standard deviation, RSD R (%) | 35 | 18 | 22 | 39 | 12 | 35 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,1 | 0,1 | 0,1 | 0,5 | 0,6 | 0,7 |
Analyte | Vanillin | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 16 | 15 | 16 | 16 | 16 | 16 |
No of results accepted (laboratories) | 16 | 15 | 16 | 16 | 16 | 16 |
Mean value (mg/l) | 0,5 | 0,2 | 1,2 | 1,2 | 3,2 | 3,9 |
Repeatability standard deviation, s r (mg/l) | 0,03 | 0,02 | 0,06 | 0,11 | 0,11 | 0,09 |
Repeatability relative standard deviation, RSD r (%) | 6,8 | 9,6 | 4,6 | 8,9 | 3,5 | 2,3 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,1 | 0,1 | 0,2 | 0,3 | 0,3 | 0,3 |
Reproducibility standard deviation, s R (mg/l) | 0,09 | 0,06 | 0,18 | 0,27 | 0,41 | 0,62 |
Reproducibility relative standard deviation, RSD R (%) | 19 | 25 | 15 | 22 | 13 | 16 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,3 | 0,2 | 0,5 | 0,8 | 1,2 | 1,7 |
Analyte | Syringaldehyde | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 16 | 15 | 16 | 16 | 16 | 16 |
No of results accepted (laboratories) | 13 | 13 | 13 | 12 | 14 | 13 |
Mean value (mg/l) | 1,0 | 0,2 | 4,8 | 3,2 | 10,5 | 9,7 |
Repeatability standard deviation, s r (mg/l) | 0,03 | 0,02 | 0,04 | 0,08 | 0,10 | 0,09 |
Repeatability relative standard deviation, RSD r (%) | 2,6 | 8,1 | 0,8 | 2,6 | 0,9 | 0,9 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,1 | 0,1 | 0,1 | 0,2 | 0,3 | 0,3 |
Reproducibility standard deviation, s R (mg/l) | 0,08 | 0,07 | 0,23 | 0,19 | 0,39 | 0,43 |
Reproducibility relative standard deviation, RSD R (%) | 8 | 33 | 5 | 6 | 4 | 4 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,2 | 0,2 | 0,7 | 0,5 | 1,1 | 1,2 |
Analyte | Coniferaldehyde | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 13 | 12 | 13 | 12 | 13 | 13 |
No of results accepted (laboratories) | 12 | 12 | 13 | 12 | 13 | 13 |
Mean value (mg/l) | 0,2 | 0,2 | 0,6 | 0,8 | 4,6 | 1,3 |
Repeatability standard deviation, s r (mg/l) | 0,02 | 0,02 | 0,03 | 0,03 | 0,09 | 0,06 |
Repeatability relative standard deviation, RSD r (%) | 9,2 | 9,8 | 4,6 | 4,3 | 1,9 | 4,5 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,04 | 0,04 | 0,07 | 0,09 | 0,24 | 0,16 |
Reproducibility standard deviation, s R (mg/l) | 0,04 | 0,04 | 0,11 | 0,18 | 0,38 | 0,25 |
Reproducibility relative standard deviation, RSD R (%) | 23 | 27 | 21 | 23 | 8 | 19 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,1 | 0,1 | 0,3 | 0,5 | 1,1 | 0,7 |
Analyte | Sinapaldehyde | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 14 | 14 | 14 | 14 | 15 | 14 |
No of results accepted (laboratories) | 14 | 13 | 12 | 13 | 13 | 12 |
Mean value (mg/l) | 0,3 | 0,2 | 0,2 | 1,6 | 8,3 | 0,3 |
Repeatability standard deviation, s r (mg/l) | 0,02 | 0,01 | 0,02 | 0,06 | 0,14 | 0,03 |
Repeatability relative standard deviation, RSD r (%) | 7,5 | 4,6 | 11,2 | 3,7 | 1,6 | 11,4 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,06 | 0,03 | 0,06 | 0,17 | 0,38 | 0,08 |
Reproducibility standard deviation, s R (mg/l) | 0,09 | 0,05 | 0,08 | 0,20 | 0,81 | 0,18 |
Reproducibility relative standard deviation, RSD R (%) | 31 | 27 | 46 | 13 | 10 | 73 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,2 | 0,2 | 0,2 | 0,6 | 2,3 | 0,5 |
Analyte | Gallic acid | |||||
---|---|---|---|---|---|---|
Sample | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 16 | 15 | 16 | 16 | 16 | 16 |
No of results accepted (laboratories) | 15 | 14 | 16 | 16 | 16 | 16 |
Mean value (mg/l) | 1,2 | 0,4 | 2,0 | 6,1 | 7,3 | 21,8 |
Repeatability standard deviation, s r (mg/l) | 0,07 | 0,04 | 0,06 | 0,18 | 0,18 | 0,60 |
Repeatability relative standard deviation, RSD r (%) | 6,1 | 8,1 | 2,9 | 3,0 | 2,4 | 2,8 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,2 | 0,1 | 0,2 | 0,5 | 0,5 | 1,7 |
Reproducibility standard deviation, s R (mg/l) | 0,43 | 0,20 | 0,62 | 3,3 | 2,2 | 7,7 |
Reproducibility relative standard deviation, RSD R (%) | 36 | 47 | 31 | 53 | 30 | 35 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 1,2 | 0,6 | 1,7 | 9,1 | 6,2 | 21,7 |
Analyte | Ellagic acid | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 7 | 7 | 7 | 7 | 7 | 7 |
No of results accepted (laboratories) | 7 | 7 | 7 | 7 | 7 | 6 |
Mean value (mg/l) | 3,2 | 1,0 | 9,5 | 13 | 13 | 36 |
Repeatability standard deviation, s r (mg/l) | 0,20 | 0,16 | 0,30 | 0,41 | 0,95 | 0,34 |
Repeatability relative standard deviation, RSD r (%) | 6,3 | 16 | 3,2 | 3,2 | 7,4 | 1,0 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,6 | 0,4 | 0,9 | 1,1 | 2,7 | 1,0 |
Reproducibility standard deviation, s R (mg/l) | 1,41 | 0,42 | 4,0 | 5,0 | 4,9 | 14 |
Reproducibility relative standard deviation, RSD R (%) | 44 | 43 | 42 | 39 | 39 | 40 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 4,0 | 1,2 | 11 | 14 | 14 | 40 |
Analyte | Vanillic acid | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 15 | 15 | 15 | 15 | 15 | 15 |
No of results accepted (laboratories) | 12 | 11 | 14 | 14 | 15 | 14 |
Mean value (mg/l) | 0,2 | 0,2 | 1,5 | 0,8 | 2,4 | 2,7 |
Repeatability standard deviation, s r (mg/l) | 0,03 | 0,04 | 0,03 | 0,10 | 0,13 | 0,21 |
Repeatability relative standard deviation, RSD r (%) | 14,2 | 16,5 | 2,3 | 12,6 | 5,3 | 7,7 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,1 | 0,1 | 0,1 | 0,3 | 0,4 | 0,6 |
Reproducibility standard deviation, s R (mg/l) | 0,06 | 0,05 | 0,51 | 0,2 | 1,22 | 0,70 |
Reproducibility relative standard deviation, RSD R (%) | 28 | 20 | 35 | 31 | 51 | 26 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,2 | 0,1 | 1,4 | 0,7 | 3,4 | 2,0 |
Analyte | Syringic acid | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 16 | 15 | 16 | 16 | 16 | 16 |
No of results accepted (laboratories) | 16 | 15 | 15 | 15 | 16 | 15 |
Mean value (mg/l) | 0,4 | 0,2 | 2,5 | 1,4 | 3,4 | 4,8 |
Repeatability standard deviation, s r (mg/l) | 0,03 | 0,02 | 0,06 | 0,13 | 0,08 | 0,11 |
Repeatability relative standard deviation, RSD r (%) | 6,7 | 12,6 | 2,3 | 9,0 | 2,3 | 2,3 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,1 | 0,1 | 0,2 | 0,4 | 0,2 | 0,3 |
Reproducibility standard deviation, s R (mg/l) | 0,08 | 0,05 | 0,29 | 0,26 | 0,43 | 0,67 |
Reproducibility relative standard deviation, RSD R (%) | 19 | 29 | 11 | 18 | 13 | 14 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,2 | 0,1 | 0,8 | 0,7 | 1,2 | 1,9 |
Analyte | Scopoletin | |||||
---|---|---|---|---|---|---|
Samples | Whisky | Brandy | Rum | Cognac 1 | Bourbon | Cognac 2 |
No of laboratories participating | 10 | 10 | 10 | 10 | 10 | 10 |
No of results accepted (laboratories) | 9 | 8 | 9 | 8 | 8 | 8 |
Mean value (mg/l) | 0,09 | 0,04 | 0,11 | 0,04 | 0,65 | 0,15 |
Repeatability standard deviation, s r (mg/l) | 0,0024 | 0,0008 | 0,0018 | 0,0014 | 0,0054 | 0,0040 |
Repeatability relative standard deviation, RSD r (%) | 2,6 | 2,2 | 1,6 | 3,3 | 0,8 | 2,7 |
Repeatability limit, r (mg/l) (r = 2,8 × s r ) | 0,007 | 0,002 | 0,005 | 0,004 | 0,015 | 0,011 |
Reproducibility standard deviation, s R (mg/l) | 0,01 | 0,01 | 0,03 | 0,01 | 0,09 | 0,02 |
Reproducibility relative standard deviation, RSD R (%) | 15 | 16 | 23 | 17 | 15 | 15 |
Reproducibility limit, R (g/l) (R = 2,8 × s R ) | 0,04 | 0,02 | 0,07 | 0,02 | 0,26 | 0,06 |
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