Application Note

No. 065/2011
Total SO2 in Wine

Distillation Unit K-355:

Determination of Total SO2 in Wine
A Comparison with the OIV SO2 Method
1. Introduction

The determination of Total SO2 contents in wine poses a challenge to steam distillation because
SO2 is withheld in adducts and is only slowly released when boiled. Recognized results are
obtained by means of the generally accepted OIV SO2 Method [1] which as a drawback is
complicated to handle and takes a long time for completion. The BUCHI SO2 Method presented
in this report is trouble-free and can easily be carried out in half of the time (Table 1).

Table 1: Time needed for one determination

Apparatus Distillation [min] Titration [min]
BUCHI K-355 5-6 5
OIV SO2 15 5

The new method is based on acidification of the wine samples with an acid mixture of methanol,
water and ortho-phosphoric acid to release SO2 efficiently from the chemical equilibrium during
distillation. The BUCHI SO2 absorption glass consists of two connected flasks, a first bottle
containing an iodine standard solution and a second one to which ethanol is added. The
determination is carried out by means of a redox back titration of the iodine solution with
standard thiosulfate solution. A unit specific calibration is needed. In a linear correlation slope
and intercept of the plot of expected SO2 amounts versus measured SO2 are determined
leading to a linear correlation equation which is applied to the calculation of SO2 contents in
i
wines. The linear calibration equation has proven to be stable over a period of months .

2. Equipment

⋅ Distillation Unit K-355 with SO2 absorption glass (order number 043070)
⋅ Glassware according to OIV [1]
⋅ Metrohm DMP 785 Titrino
⋅ Metrohm Pt-Titrode 6.0431.100
⋅ Volumetric pipette 5 mL
⋅ Glass beaker 500 mL

3. Chemicals and Materials

Chemicals:
⋅ Ethanol purum, Fluka (02875)
⋅ Sodium sulfite, Sigma-Aldrich (71989)
⋅ Citric acid monohydrate, Fluka (27490)
⋅ Ethanal (acetaldehyde), Fluka (00070) (store in refrigerator)
⋅ Ethanal solution: Pipette 1-1.3 mL ethanal into a volumetric flask of 1 L which contains
approx. 100 mL of deionized water. Fill up to the mark with deionized water.
⋅ Stabilized standard stock solution of 160 mg SO2/L: 120 mL ethanol, 323.0 mg Na2SO3,
2.03 g citric acid monohydrate and 320 mL of ethanal solution are placed into a volumetric
flask of 1 L and filled up to the mark with deionized water.
⋅ 12 % ethanol solution: Pour 120 mL of ethanol into a 1 L volumetric flask and fill up to the
mark.
⋅ Iodine standard solution 0.05 mol/L, Riedel-de Haën (35090)
⋅ 0.01 mol/L Na2S2O3 standard solution, Riedel-de Haën (38243)
⋅ Ortho-phosphoric acid (85%), Riedel-de Haën (38243)
⋅ Methanol, Sigma Aldrich (32213)
⋅ Acid mixture: For the preparation of approx. 1 liter mix 500 mL methanol, 400 mL of
deionized water and 50 mL o-phosphoric acid in a measuring cylinder of 1 L.
⋅ Sulfuric acid 0.5 mol/L, Riedel-de Haën (35354)

For a safe handling please pay attention to all corresponding MSDS!

Application Note 065/2011 March 2011 2/14

 Samples:

Table 2: Wine samples used for SO2 determinations (F = France, CH = Switzerland, USA = United States, I = Italy,
P = Portugal, E = Spain)
ii
Wine No Name of wine Origin Type Alcohol [% (v/v) ]
1 Chablais CH (Vaud) red 12.5
2 Salvagnin CH (Vaud) red 12.5
3 Vin de table P red 12.0
4 Padre Alonso E (Navarro) red 12.5
5 La Tassée du Roy F rosé 12.5
6 Côtes-du-Rhône F red 13.0
7 Cabernet Sauvignon USA (California) red 13.5
8 Montepulciano I (Abbruzzo) red 12.0
9 Château d'Allaman CH (VD white 12.0
10 Mont-sur-Rolle CH (VD) white 11.8
11 Muscat + excess SO2 F (Pyrénées) white 15.2

4. Procedure

The determination of SO2 in wine includes the following steps:

⋅ Comparison of sample volumes and concentrations
⋅ Adjusting the Distillation Unit
⋅ Distillations and Titrations

4.1. Comparison of sample volumes and concentrations

In Table 3 a comparison of sample volumes Vsample and concentrations c(SO2) is given for the
OIV SO2 and the BUCHI SO2 Method. Due to the amount of stabilized SO2 standard solution
used, the results in this report are valid in the range of W(SO2)standard =1– 8 mg SO2/sample
corresponding to 5 – 50 mL SO2 standard stock solution of 159.4 mg SO2/L, see Table 4. The
smallest two standards in Table 4 show significantly larger rsd's compared to the following ones
which is due to SO2 amounts below or close to the limit of quantification [2])

Table 3: Range of Total SO2 concentrations covered by given volume of pipetted sample for the OIV SO2
instrumentation and for steam distillation by BUCHI distillation unit. Calibration range W(SO2)calib = 1 – 8 mg SO2 (see
Fig. 2).

c(SO2)OIV [mg SO2/L] Vsample OIV [mL] c(SO2)BUCHI [mg SO2/L] Vsample BUCHI [mL]
≤ 50 mgSO2/L 50 30-80 100
≥ 50 mgSO2/L 20 50-160 50

130-300 25

Application Note 065/2011 March 2011 3/14

4.2. Adjusting the Distillation Unit
It is recommended to use the BUCHI SO2 absorption glass to collect the distillate (Fig. 1).

Fig. 1: Receiving vessels for collecting SO2 in aqueous iodine solution.

4.2.1. Quality Check of Iodine Solution

The volume of freshly prepared 0.01 mol/L thiosulfate standard solution for the direct titration of
5 mL 0.05 mol/L iodine solution is determined. A typical value for fresh iodine standard is in the
range of 49.5 to 50 mL and the deviation should be <0.7 mL. If during an experiment the
thiosulfate solution is to be renewed repeat the quality check to ensure identical conditions for
the determinations before and after changing the titrant solution.

4.2.2. Conditioning the Distillation Unit K-355

The distillation unit K-355 is conditioned by distilling approx. 50 mL of methanol at a steam
power of 100% for 6 minutes.

4.2.3. Adjusting Distillation Time

Immediately after the conditioning of the K-355 the distillation time needed to fill the first
receiving vessel completely with distillate has to be determined. With the BUCHI SO2 absorption
glass on the distillation unit K-355 a typical distillation time would be in a range of 5-6 minutes.
For the adjustment add 50 mL of acid mixture and 50 mL of 12% ethanol solution into the
sample tube. Pipette 5 mL 0.05 mol/L iodine solution and 30 mL of water to the first and 30 mL
ethanol to the second receiving vessel. Set the distillation time to 6:00 minutes and determine
the time needed to fill the first receiver. Apply this time to the blank distillations and for the
samples. The optimal distillation time is specific to a distillation unit and only needs to be
determined initially.

4.3. Distillations and Titrations

4.3.1. Blanks
Start with the blank determination immediately after the conditioning or adjustment steps
described above. By means of a volumetric pipette add 5 mL of 0.05 mol/L iodine solution into
the first receiving vessel of the BUCHI SO2 absorption glass. Add 30 mL of deionized water to

Application Note 065/2011 March 2011 4/14

the first vessel and 30 mL of ethanol to the second. Insert glass stopper connecting the two
vessels and make sure the ends of the two plastic tubes are immersed into the liquids. Connect
the SO2 absorption glass to the outlet of the cooler. Pour a volume of 12 % (v/v) ethanol solution
corresponding to the volume of wine sample and add 50 mL of acid mixture. Attach the sample
tube to the K-355 and distill for the adjusted distillation time (e.g. 5:40 minutes) using a steam
power of 100%. Make sure the same distillation times are applied for blanks and samples.
Combine the contents of the two receiving vessels in a 500 mL beaker and carefully rinse the
stopper and the two vessels three times with deionized water. Acidify the combined distillates
with 2 mL of 0.5 mol/L sulfuric acid and titrate with 0.01 mol/L thiosulfate standard solution.
Typical titrant consumptions for the blanks are in a range of 47 – 49 mL and the standard
deviation should not exceed sd > 0.7 mL. Use the average of 2-3 blank determinations meeting
these conditions for the calculations.

4.3.2. Calibration with the stabilized SO2 standard stock solution

In this report the calibration is based on different volumes of a stabilized SO2 standard solution
for which the concentration of the standard stock solution was determined by means of the OIV
iii
SO2 Method: c(SO2)OIV = 159.4 mg SO2/L .

The amounts of SO2 associated with the corresponding added volumes of standard stock
solutions W(SO2)standard are calculated by means of equations (1) and (2).

Table 4: Correlation of VTcalib, W(SO2)calib and W(SO2)standard as a function of the respective volumes Vstandard of the
stabilized SO2 standard stock solution, VTblank = 47.855 mL, (n = 3), c(SO2)stock determined by OIV SO2 Method to 159.4
mg SO2/ L, (n = 2), (Standard deviations of VTcalib and relative standard deviations of W(SO2)calib in italics).
T
Vstandard [mL] V calib [mL] W(SO2)calib [mg] W(SO2)standard [mg] rsd for n=2 [%]
5 45.988 (0.202) 0.60 (10.8 %) 0.80 10.8
10 43.369 (0.249) 1.44 (5.5 %) 1.59 5.5
15 41.134 (0.083) 2.15 (1.8 %) 2.39 1.2
20 39.174 (0.124) 2.78 (1.4 %) 3.19 1.4
25 36.639 (0.170) 3.59 (1.5 %) 3.99 1.5
50 25.291 (0.120) 7.23 (0.5 %) 7.97 0.5

By means of a volumetric pipette add 5 mL of 0.05 mol/L iodine solution into the first receiving
vessel of the BUCHI SO2 absorption glass. Add 30 mL of deionized water to the first receiving
vessel and 30 mL of ethanol to the second one. Insert glass stopper connecting the two vessels
and make sure the ends of the two plastic tubes are immersed into the liquids. Connect the SO2
accessory to the outlet of the cooler. In consecutive distillations and titrations pipette 6-7
different volumes in a range of 5 to 50 mL stabilized SO2 standard solution to the sample tube
and add 50 mL of acid mixture. Compensate for the difference between 50 mL and the used
volumes of standards by adding 12% (v/v) ethanol solution, e.g. 10 mL standard + 40 mL 12%
(v/v) ethanol solution.

Attach the sample tube to the K-355 and distil for the adjusted distillation time. Typically two
distillations are sufficient but it is recommended to add distillations if the relative standard
deviations are >3.5%.

Application Note 065/2011 March 2011 5/14

 Calibration of SO2 Determination Using SO2-Standard of 159.4 mgSO2/l

9.00
y = 1.0894x + 0.0909
8.00 2
R = 0.9996

W(SO 2)standard [mgSO 2]

7.00
6.00
5.00
4.00
3.00
2.00
1.00
0.00
0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00
W(SO2)calib [mgSO2]

Fig. 2: Calibration curve for the determination of Total SO2 and linear equation correlating the measured SO2
amounts (x-axis) to the theoretical SO2 contents (y-axis).

The linear regression leads to a linear equation with a slope a = 1.0894 and an intercept of
0.0909 mg SO2 (Fig. 2).

4.3.3. Wine Samples

Proceed as described for the standards but use the appropriate volumes of wine sample as
given in Table 3. OIV conformal results of SO2 in wine are calculated applying the linear
equation (5) based on the correlation data given in Table 4.

5. Calculation

mstandard • M(SO2 ) (1)

c(SO2 )stock =
M (Na2SO3 ) • Vstock

c(SO2 )stock • Vstandard (2)

W(SO2 )standard =
1000

T T
(Vblank - Vcalib ) • c T • M(SO2 ) (3)
W(SO2 )calib =
z

T T
(Vblank - Vsample ) • c T • M(SO2 )
W(SO2 )sample = (4)
z

W(SO2 )sample • 1000

c(SO2 )sample = (5)
Vsample

W(SO2 ) = a • W(SO2 )sample + b (6)

W(SO2 ) • 1000
c(SO2 ) = (7)
Vsample

Application Note 065/2011 March 2011 6/14

 c(SO2)stock : concentration of stabilized SO2 standard [mg SO2/L]
mstandard : Weight of Na2SO3 in standard solution [g]
M(SO2) : Molar mass SO2 = 64.0648 [g/mol]
M(Na2SO3) : Molar mass Na2SO3 = 126.16 [g/mol]
Vstock : Volume of standard stock solution [L]
W(SO2)standard : Weight of SO2 calculated from Vstandard [mg SO2]
Vstandard : Volume of stabilized SO2 standard [mL]
W(SO2)calib : Determined weight of SO2 in standard calibration [mg SO2]
T
V blank : Consumption of thiosulfate solution for blank [mL]
T
V calib : Consumption of thiosulfate solution for standard [mL]
T
c : Concentration of thiosulfate standard [mol/L]
z : Redox valency of thiosulfate = 2
W(SO2)sample : Determined weight of SO2 in sample [mg SO2]
T
V sample : 0.01 mol/L thiosulfate consumption for sample [mL]
c(SO2)sample : Determined SO2 concentration in sample (by BUCHI SO2 method) [mg SO2/L]
Vsample : Sample volume [mL]
c(SO2)OIV : Determined SO2 concentration in sample (by OIV SO2 method) [mg SO2/L]
W(SO2) : Adjusted weight of SO2 [mg SO2]
a : slope of linear regression
b : Intercept of linear regression [mg SO2]
c(SO2) : Adjusted SO2 concentration [mg SO2/L]

BUCHI offers an Excel template for the calculations for the redox back-titration of iodine with
thiosulfate. Examples are given in chapter '6 Results' with the equations (3) – (7) used in the
calculations.

6. Results

Adjusted SO2 concentrations c(SO2)adj found in 11 wine samples by means of the BUCHI SO2
Method are compared with the corresponding results c(SO2)OIV obtained by the OIV SO2
Method. The volumes of sample were chosen in accordance with Table 3.
In Tables 6-16 the calculations of results is given for the individual SO2 determinations using the
linear correlation shown in Fig. 2. The calculations are based on equations (4) – (7).

Table 5: Results of Total SO2 determinations using the K-355 for 11 wines (for OIV- and BUCHI SO2 Methods: n=2, for
BUCHI SO2 Method: Montepulciano n=3, standard deviations are given in italics).
Name of wine Vsample [mL] c(SO2)OIV [mg SO2/L] c(SO2) [mg SO2/L]
iv
Chablais 50 38.2 (0.0) 39.4 (1.1)
Salvagnin 50 63.2 (0.2) 64.1 (0.7)
Vin de table 50 67.7 (0.2) 68.8 (0.7)
Padre Alonso 50 68.6 (0.2) 72.3 (0.3)
La Tassée du Roy 50 85.6 (0.2) 87.5 (0.3)
Côtes-du-Rhône 50 88.6 (1.2) 90.1 (0.6)
Cabernet Sauvignon 50 89.1 (0.2) 91.8 (1.1)
Montepulciano 50 93.6 (0.7) 94.8 (0.4)
Château d'Allaman 50 101.6 (0.2) 102.6 (2.4)
Mont-sur-Rolle 50 103.2 (0.2) 105.3 (0.6)
Muscat + excess SO2 25 212.0 (1.1) 220.9 (2.7)

Application Note 065/2011 March 2011 7/14

 Comparison Total SO2

250

200

150
mgSO 2/l

100

50

0
1 2 3 4 5 6 7 8 9 10 11

Wine No

Fig. 3: Graphical comparison of the OIV SO2 Method (left column in red) to the BUCHI SO2 Method (right
column in yellow) for the wines numbered 1 to 11. Experimental repeatabilities r = 2.8*standard deviation are
indicated as error limits.

In the captions of the tables below short names of the wines are given the way as they were
introduced in Tables 2 and 3 and if appropriate in brackets complemented with the more
complete names used in the captions of tables for data evaluation.

Table 6: Determination of Total SO2 contents in wine 4 Chablais (= Chablais District d'Aigle)

Application Note 065/2011 March 2011 8/14

Table 7: Determination of Total SO2 contents in wine 2 Salvagnin (= Salvagnin Rouge LE PAON)

Table 8: Determination of Total SO2 contents in wine 6 Vin de Table (= Rouge du Portugal)

Application Note 065/2011 March 2011 9/14

Table 9: Determination of Total SO2 contents in wine 7 Padre Alonso

Table 10: Determination of Total SO2 contents in wine 8 La Tassée du Roy

Application Note 065/2011 March 2011 10/14

Table 11: Determination of Total SO2 contents in wine 1 Côtes-du-Rhône

Table12: Determination of Total SO2 contents in wine 3 Cabernet Sauvignon (= Cabernet Sauvignon RM)

Application Note 065/2011 March 2011 11/14

Table 13: Determination of Total SO2 contents in wine 5 Montepulciane (= Montepulciano d'Abruzzo)

Table 14: Determination of Total SO2 contents in wine 10 Château d'Allaman

Application Note 065/2011 March 2011 12/14

Table 15: Determination of Total SO2 contents in wine 9 Mont-sur-Rolle)

Table 16: Determination of Total SO2 contents in wine 11 Muscat + excess SO2 (= Muscat de Rivealtes + Meta-Bisulfite)

Application Note 065/2011 March 2011 13/14

7. Comparison to Standard Methods

The International Organization of Vine and Wine (Organization Internationale de la Vigne et du

Vin = OIV) has published the 'Compendium of International Methods of Wine and Must Analysis'
which provides standard procedures for various constituents in Wine including the widely
accepted standard for the analysis of Total SO2 in wine [1]. In Table 17 the OIV SO2 Method is
compared to the BUCHI SO2 Method.

Table 17: Comparison of the OIV SO2 Method to the BUCHI SO2 Method
Deviation OIV SO2 Method BUCHI SO2 Method
SO2 entrainment N2-stream Steam
Acidification phosphoric acid Methanol / H2O / phosphoric acid
Distillation 15 min 5-7 min
Solution in receiver H2O2 Aqueous I2-solution
Titration type Acid-base Redox back titration
Titrant NaOH Thiosulfate
Titrated analyte H2SO4 I2

8. Conclusion

The new BUCHI SO2 Method for the determination of Total SO2 in wine reproduces the SO2
result obtained by means of the OIV SO2 Method which to date is the widely accepted standard
method.

9. Acknowledgements

The author would like to express his gratitude to Mr. Kay Pfluger for his greatly appreciated
support in carrying out Total SO2 determinations by the OIV SO2 Method at the Labs of
SCHENK SA in Rolle.

10. References

[1] Compendium of International Methods of Wine and Must Analysis, Edition 2011, Volume
2, OIV-MA-AS323-04A and OIV-MA-AS323-04B.
[2] Determinations of Limit of Detection, Limit of Quantification and Recovery Rates,
Application No.K-355_011 V.1.1 (2008)
Operation Manual Distillation Unit K-355

i
By long duration tests carried out on K-355’s at BUCHI and with SCHWENK SA.
ii
According to label
iii
In a related report AN 066/2011 the amounts of SO2 in the determined standard solutions
were based on the weight of Na2SO3 weighed in when the stock solution was prepared.
iv
See Table 2: A volume of 100 mL would have been possible also

Application Note 065/2011 March 2011 14/14