Food Control 16 (2005) 177–181

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Methanol contents of Turkish varietal wines and effect of processing

Turgut Cabaroglu *

Department of Food Engineering, Faculty of Agriculture, University of Cukurova, 01330 Adana, Turkey
Received 18 September 2003; received in revised form 3 January 2004; accepted 13 January 2004

Abstract
Methanol levels of Turkish varietal wines obtained from different representative viticultural regions were determined and the
effect of maceration time and pectolytic enzyme treatments on the methanol levels of the red wines were investigated. 164 wine
samples (60 white, 4 ros
e, 100 red) were analysed by gas chromatography–flame ionisation detector (GC–FID). Methanol levels
ranged from 30.5 to 121.4 mg/l in white wines, 62.5 to 84.6 mg/l in ros
e, and 61.0 to 207.0 mg/l in red wines. The highest level was
€ uzg€
found in Sultaniye among the white wines, and in Ok€ oz€
u among the red wines. Methanol content of red wines increased with
maceration time. Pectolytic enzyme treatments produced significantly higher methanol levels than the control wine. All the results
show that methanol levels of Turkish wines are under the maximum acceptable limits of International Office of Vine and Wine (OIV)
and do not represent a risk to consumer health.

2004 Elsevier Ltd. All rights reserved.

Keywords: Methanol; Maceration time; Pectolytic enzyme; Turkish wines; Legislation

1. Introduction quantities, fairly large amounts of methanol have been

Methanol, so-called methyl alcohol (CH3 OH) is a
colourless, liquid at ambient temperatures with a bland
odour (Gnekow & Ough, 1976). It is well known that
methanol is a toxic and harmful substance to human
health (when taken orally at 340 mg/kg of body weight)
of whose ingestion or inhalation can cause blindness or
death (Gnekow & Ough, 1976; Toxic substance list,
1973). Following ingestion, it is oxidised to formic acid,
both toxic to the central nervous system. Formic alde-
hyde deteriorates the optical nerve, causing blindness
(Ribereau-Gayon, Glories, Maujean, & Dubourdieu,
2000). Newsholme and Leech (1986) reported that oxi-
dised methanol produces lactic acidosis which is a met-
abolic disease caused by an increase in blood levels of
lactic acid. Its symptoms lead to weakness, vomiting and
finally coma and death. The level of 200 mg/l is con-
sidered the maximum safe concentration to be inhaled
for a continuous eight-hour working day. The lethal
dose varies between 60 and 250 ml (Gnekow & Ough,
1976).
Nearly all-alcoholic beverages, contain methanol.
Although methanol is present in beers in very small
*
Fax: +90-322-338-61-73.
E-mail address: tcabar@cu.edu.tr (T. Cabaroglu).
0956-7135/$ - see front matter
2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodcont.2004.01.008
found in wines and distilled beverages (Nykanen &
Suomalainen, 1983). According to implementation of
the hazard analyses critical control point (HACCP)
system in winemaking, methanol content is one of the
critical control point parameters that should be con-
trolled during the alcoholic fermentation stage to pro-
duce wine safe for human consumption (Kourtis &
Arvanitoyannis, 2001). Methanol found in wines is
formed from enzymatic degradation of natural pectic
substances (pectin) present in crushed grapes by pec-
tinesterase (Cordonnier, 1987; Ough & Amerine, 1988;
Ribereau-Gayon et al., 2000). The formation of meth-
anol in wines is dependent upon a number of factors
such as the grape variety (especially grape skins which
contain a high pectin content), grape health, maceration
treatment, fermentation temperature and pectolytic en-
zyme treatment. Although numerous studies have been
done on the methanol content of wines in several wine
producing countries such as France, Italy, Spain, Por-
tugal, USA, Germany, Albania (Nykanen & Suomalai-
nen, 1983) no data have been reported on the methanol
content of Turkish wines.
Ribereau-Gayon, Peynaud, Sudraud, and Ribereau-
Gayon (1982) reported that red wines contain a higher
level of methanol than roses, while white wines contain
even less. Because of the higher pectin content of hybrid
grape skins, wines made from hybrids have a higher
178 T. Cabaroglu / Food Control 16 (2005) 177–181
methanol content than those made from Vitis vinifera
(Ribereau-Gayon et al., 2000). Flanzy and Bouzigues
(1959) showed that maceration produced more metha-
nol as a function of contact time between juice and
grape solids. Some authors have found that the use of
pectolytic enzymes to facilitate extraction or clarifica-
tion of the must, induced an increase in wine methanol
content (Brown & Ough, 1981; Gnekow & Ough, 1976;
Revilla & Gonzales-SanJose, 1998). Findings from pre-
vious studies showed that methanol levels of wines were
generally under the maximum limits (150 mg/l for white
and ros
e, 300 mg/l for red) given by International Office
of Vine and Wine (OIV). However, levels above the
given limits were reported by some authors (Cordonnier,
1987; Gnekow & Ough, 1976; Lee, Robinson, Van Bu-
ren, Acree, & Stoewsand, 1975; Nykanen & Suomalai-
nen, 1983).
This study was mainly carried out to evaluate meth-
anol content of Turkish varietal wines obtained from
several viticultural regions, and secondly, to investigate
the effect of maceration time and pectolytic enzyme
treatments on the methanol levels of the experimental
wines.
2. Materials and methods
2.1. Wine samples
To measure the content of methanol in Turkish wine,
164 representative varietal wine samples (60 white, 4
ros
e, 100 red) of bottled (750 ml each) obtained from
main viticultural regions were selected. Samples were
mainly collected from eleven different wineries and
purchased from the marketplace. In addition, some
experimentally produced samples (explained below) at
the experimental winery of Department of Food Engi-
neering were also used.
2.2. Maceration and pectolytic enzyme treatments
Maceration treatment: For the maceration treatment
three of the most popular red wine grapes Ok€ € uzg€oz€u
(2250 kg), Bo gazkere (2250 kg) from Eastern Anatolia
and Kalecik karası (1200 kg) from Central Anatolia
were used. Undamaged healthy grapes of each variety
were manually harvested at optimum maturity and
transported by lorry to the experimental winery. Grapes
were de-stemmed and crushed on a grape destemmer–
crusher. Three different maceration times were con-
ducted for each variety. Each variety was divided into
three lots to carry out the following maceration times: 3,
6 and 10 days. Then, each lot was transferred into
stainless-steel tanks for the maceration and 40 mg/kg
sulphur dioxide was added. Alcoholic fermentation was
conducted with 0.2 g/l Fermirouge-7303 (Gist Brocades-
France) at 25 C. Fermentation rate was measured by
densitymeter. After the maceration of each lot, the must
was pressed gently, and the free-run and pressed juice
were combined. At the end of alcoholic fermentation,
the wines were racked and kept at 20 C to stimulate
malolactic fermentation. Upon completion of malolactic
fermentation, wines were racked and stored at 15 C
during one year before methanol analysis.
Pectolytic enzyme treatment: For enzyme treatment,
€ uzg€
Ok€ oz€
u grapes (2250 kg) from Eastern Anatolia were
used. Healthy grapes were harvested and transported to
the experimental winery. Two different colour extracting
pectolytic enzymes were used. Their codes and applica-
tion doses suggested by producers were: Peclyve-V
(Lyven) 0.02 g/l (P.-V Ex.), Rapidase Ex.-Colour (Gist
Brocades) 0.03 g/l (R. Ex-C.). Grapes were randomly
divided into three equal lots. Each lot was de-stemmed,
crushed and 40 mg/l sulphur dioxide was added. After 1
h, P.-V Ex. and R. Ex-C. were added into second and
third lots respectively. First lot was used as a control
with no enzyme addition. Following the enzyme addi-
tion, each must was inoculated with commercial yeast,
0.2 g/l Fermirouge-7303 (Gist-Brocades-France). The
alcoholic fermentation on the skins (maceration) was
carried out at 25 C during 8 days. After that, the same
procedures were applied on the wine as explained above.
2.3. Methanol analysis
Methanol content was determined by gas chroma-
tography (GC) on distilled wine samples by adding an
internal standard according to reference method of OIV
(1990).
Distillation: 100 ml of each wine sample was distilled
by a simple distillation equipment as explained by OIV;
100 ml of each distillate was collected in a volumetric
flask at low temperature to prevent losses. Each sample
was distilled in triplicate. A standard solution of meth-
anol (Merck) was distilled in the same way to determine
the recovery of methanol, which was 99.2%. Distilled
sample of 100 ml was added with 10 mg of 4-methyl-2-
pentanol (Merck) as internal standard. One microlitre
sample was injected into the GC.
GC condition: GC used was Shimadzu 14B chro-
matograph equipped with a fused capillary column
coated with Permabond CW20M (30 m · 0.25 mm
i.d. · 0.25 m film thickness, Macherey-Nagel, USA) and
flame ionisation detector (FID). Injections were made in
split mode (split ratio 10:1). Injection port temperature
was 200 C and the oven was programmed from 70 to
150 C at a rate of 4 /min. The FID temperature was
250 C. The carrier gas was helium with a flow rate of 55
kPa. Each sample was injected in triplicate. The stan-
dard solution of methanol (in 10% ethanol) containing
internal standard at various concentration (30–400 mg/l)
were analysed to calculate the response factor.
 T. Cabaroglu / Food Control 16 (2005) 177–181 179

The methanol concentration of each wine sample is
calculated by internal standard method with respect to
the internal standard from response factor (Kelly et al.,
1999).
The statistical analysis of the data obtained from
maceration and pectolytic enzyme treatments were car-
ried out by an analysis of variance (one-way Anova).
Means between control and treated wines were com-
pared at P < 0:05 by Fisher’s least significant difference
test using the SPSS 10.0 for Windows statistical pack-
age.
3. Results and discussion
3.1. Composition of wine samples
To evaluate methanol contents of Turkish varietal
wines representative 60 white, 4 ros
e and 100 red wines
produced by wine companies and experimentally were
analysed and the results are summarised in Table 1 by
giving minimum, maximum and average concentration
of each varietal wine.
The methanol levels of white wines ranged from 30.5
to 121.4 mg/l, with an average value of 58.5 mg/l for 60
samples. The methanol content of white wines from
different countries were reported as follow (Nykanen &
Suomalainen, 1983): in French wines, from 38 to 114
mg/l with an average of 60 mg/l, in Italian wines, from
zero to 182 mg/l with an average of 60 mg/l, in Portu-
guese wines from zero to 119 mg/l with an average of 63
mg/l. According to OIV (1990), maximum acceptable
level for methanol in white wines and ros
e is 150 mg/l.
This value is also the critical limit during the white wine
production as mentioned in the HACCP implementa-
tion of wine (Kourtis & Arvanitoyannis, 2001). In this
study within white wines, the highest level was detected
in a Sultaniye wine with 121.4 mg/l, which is under the
limit. According to average values of white wines, Sul-
taniye wines were also the highest, followed by Riesling,
Semillon and Emir wines. Muscat of Bornova wines
contained the lowest methanol level of all the wines
analysed. Important differences between the levels of the
same varietal wines such as Sultaniye can be explained
by the bad harvesting practices (unhealthy, overmature
grapes) or the different winemaking practices of winer-
ies.
Since the only variety was Cal karası for production
of ros
e wine in Turkey and the production was limited,
merely four samples were measured for methanol con-
tent. As can be seen from Table 1, the levels of Cal
karası wines were under the limit and slightly higher
than the levels of white wines.
Methanol contents of Turkish varietal red wines
ranged from 61.0 to 207 mg/l, with an average value of
113 mg/l for 100 samples. The methanol content of red
wines from different countries were reported as follow
(Nykanen & Suomalainen, 1983): in French wines, from
100 to 200 mg/l with an average of 163 mg/l; in Italian
wines, from zero to 635 mg/l with an average of 103 mg/
l; in Spanish wines, from 39 to 624 mg/l with an average
value of 145 mg/l; in Portuguese wines from 165 to

Table 1
Analytical data and methanol levels of Turkish varietal wine samples
Regiona Variety Colour No of samples Vintage Concentration (mg/l)
Min. Max. Average
C.A. Emir White 16 2000–2001 40.5 83.8 56.5
B.S. Narince White 16 2000–2001 38.2 72.0 49.7
Ae. Sultaniye White 8 2001 42.0 121.4 79.2
Ae. Muscat of Bornova White 6 2000–2001 30.5 52.0 45.4
M.T. Semillon White 10 2000–2001 47.7 76.3 58.8
M.T. Riesling White 4 2000 58.6 65.8 61.9
60 58.5
Ae. Cal karası Ros
e 4 2001 62.5 84.6 77.2
E.A. € uzg€
Ok€ oz€u Red 18 2000–2001 72.2 207.0 134.1
E.A. Bogazkere Red 18 2000–2001 61.0 149.6 104.4
C.A. Kalecik karası Red 16 2000–2001 88.5 152.0 118.2
M.T. Papaz karası Red 6 2001 96.3 137.0 112.6
M.T. Ada karası Red 4 2001 75.0 128.7 99.8
M.T. Cinsault Red 6 2001 85.6 130.6 106.8
M.T. Gamay Red 6 2001 78.7 120.9 102.6
Ae. Cabernet Sauvignon Red 10 2001 89.1 140.5 110.0
Ae. Carignan Red 6 2001 97.3 161.9 126.1
Ae. Merlot Red 6 2001 84.5 164.0 124.5
Ae. Alicante Bouchet Red 4 2001 94.4 129.9 112.4
100 113.7
a
C.A.: Central Anatolia, B.S.: Black sea, Ae.: Aegean, M.T.: Marmara-Thrace, E.A.: Eastern Anatolia.
180 T. Cabaroglu / Food Control 16 (2005) 177–181

264 mg/l with an average of 195 mg/l. According to OIV well known that the enzymatic system with high pec-
(1990), maximum acceptable level for methanol in red tinesterase activity degrades the pectic compounds with
wines is 300 mg/l. It can be seen from Table 1 that none methanol release.
of the red wine samples exceeded this limit. Within
varietal red wines, the highest value was found in
€ uzg€
Ok€ oz€
u wine samples with 207 mg/l. Average value of
€ 4. Conclusions
Ok€ uzg€
oz€
u wines was also the highest of all the wines
analysed (134.1 mg/l). With respect to average values,
€ uzg€ In general, 164 representative samples of varietal
following Ok€ oz€u wine the highest values found in
wines analysed showed that methanol contents of
Carignan (126.1 mg/l), Merlot (124.5 mg/l) and Kalecik
Turkish wines were under the limits given by OIV. As
karası (118 mg/l) respectively; the lowest in Ada karası
expected, white wines contained less methanol than red
(99.8 mg/l).
wines, ros
e wines possessing values between the former.
This can be explained that white grapes are directly
3.2. Effect of maceration time and enzyme treatment pressed, whereas reds are left to macerate for a certain
time period on the skins which are rich in pectin (Cor-
€ uzg€
Ok€ oz€
u, Bo
gazkere and Kalecik karası, the most donnier, 1987). It was reported that wines made from
popular red grape varieties of Turkey, were selected to white grape varieties of Vitis vinifera, Vitis labrusca and
investigate the effect of maceration time and enzyme hybrids contained less methanol than red varieties and
treatment on the methanol content. The results are generally Vitis vinifera series wines contained the lowest
shown in Table 2. The methanol level of the each vari- amounts of methanol among the group (Lee et al.,
etal wine was increased by maceration time. The levels 1975).
of Bo gazkere wines macerated for 3, 6 and 10 days were On the basis of HACCP implementation and safety
statistically different from each other. However, wine production, methanol content of wines should be
€ uzg€
Ok€ oz€
u and Kalecik karası wines macerated for 6 and under control during wine production. Taking into
10 days were different than that of macerated for 3 days. consideration the methanol levels of the analysed vari-
An important increase in the formation of methanol was etal wines, it seems that Turkish wines have met the OIV
observed during the first 6 days of maceration. limits. However, attention should be paid to Sultaniye
The use of pectolytic enzymes significantly increased € uzg€
and Ok€ oz€u wines during production, because their
the methanol levels of Ok€€ uzg€ oz€
u wines against the levels are close to maximum acceptable limits. Especially
control. Increase of methanol levels in wine made with when pectolytic enzymes are used, monitoring of
pectolytic enzymes has been reported in other studies methanol formation in wines is necessary to avoid any
(Brown & Ough, 1981; Gnekow & Ough, 1976; Revilla risk.
& Gonzales-SanJose, 1998). On the other hand signifi-
cant differences were observed between two commercial
preparations. P.-V Ex. gave a lower methanol content Acknowledgements
than the R. Ex-C. methanol content. Revilla and
Gonzales-SanJose (1998) reported that difference in the The author would like to thank to the University of
methanol production by various enzyme preparations Cukurova for financial support to this research project
could be the result of different kinds of activities. It is (project no. ZF-2002-BAP39). Many thanks to Kava-
klidere, Sevilen, Pamukkale, Kocabag, Turasan, Diren
Table 2 wine companies for wine samples.
Effect of maceration time and enzyme treatment on the methanol
levelsa (mg/l) of several Turkish wines
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3 6 10
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