Innovative Food Science and Emerging Technologies 13 (2012) 112

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Innovative Food Science and Emerging Technologies

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High hydrostatic pressure treatment of beer and wine: A review

Sencer Buzrul
Ttn ve Alkol Piyasas Dzenleme Kurumu (TAPDK), 06520, Ankara, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: High hydrostatic pressure (HHP) technology has become a reality in the food industry. Commercial use of HHP has
Received 21 May 2011 been accepted in many countries and it is possible to nd and buy products treated by HHP such as meat products,
Accepted 1 October 2011 sea foods and fruit juices. Nevertheless, no HHP-treated beer and wine are introduced in the market throughout
the world although rice wine is one of the earliest HHP-treated commercial products that appeared on the Japa-
Keywords:
nese market. This contribution compiles the studies about HHP on beer and wine: in addition to microbial destruc-
High hydrostatic pressure
Beer
tion, it has been reported that HHP improves some organoleptic properties of beer and wine without detrimental
Wine effects on important quality characteristics, such as color, pH and turbidity. Although more studies should be car-
Stabilization ried out on the sensory properties and consumer attitudes to HHP-treated beer and wine, HHP could be an alter-
Microbial inactivation native to the existing stabilization methods used in beer and wine industries.
Sensory properties Industrial Relevance: Studies have shown that HHP treatment not only inactivates the undesirable microorganisms
but also improves the organoleptic properties of beer and wine. The pressure levels used to treat beer and wine
were similar to the commercial applications used in fruit juice industry i.e., 400-600 MPa. Therefore, HHP has a
huge potential to eliminate the negative effect of heat on the aroma and avor beer and also to reduce the SO2
levels used in wine.
2011 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2. Applications of HHP on brewing process and beer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
3. Applications of HHP on wine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4. Concluding remarks and future perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

1. Introduction is dened as exposure to 60 C for 1 min. Although laboratory tests have

Beer is the most consumed alcoholic beverage in the world and its
microbial stability is achieved by heat treatment. The stability of beer
up to several months can be provided if the spoilage organisms are
destroyed. Heat treatment is either done by ash pasteurization,
where beer is rst pasteurized and then packaged aseptically usually
into metal kegs, or by tunnel pasteurization, in which beer is rst
lled into sterile glass bottles and then pasteurized through tunnel
pasteurizers (Buzrul, 2003; 2007).
Commercial rule of thumb has been to use a timetemperature rela-
tionship of 15 min at 60 C i.e., 15 pasteurization units (PU), where 1 PU
Tel.: + 90 312 218 0438; fax: + 90 312 218 0430.
E-mail addresses: sencer.buzrul@tapdk.gov.tr, sencer.buzrul@gmail.com.
indicated that values from 1 to 5 PU are effective for microbial inactiva-
tion, 830 PU are generally used, perhaps to have a built-in safety factor
in case of possible resistant organisms (Tshang & Ingledew, 1982). How-
ever, off-avors are easily formed during pasteurization since beer is a
delicate beverage. With freshness being top priority, it is evident a
method of pasteurization using no heat would be of great help to the
brewing industry (Folkes, 2004).
Unlike beer, wine cannot be treated with heat since wine charac-
teristics such as avor, taste and color are very sensitive to tempera-
ture (Mermelstein, 1998). Therefore, a common practice is the
addition of sulfur dioxide (SO2) to wine, which is added to reduce
the microbial population of the grape must and to preserve the nal
product for long period of time (Ribreau-Gayon, Dubourdieu,
Donche, & Lonvaud, 2006). SO2 acts as both an antimicrobial agent

1466-8564/$ see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ifset.2011.10.001
2 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112
and an antioxidant in wine (Amerine, Berg, & Cruess, 1967; Romano &
Suzzi, 1993). However, SO2 can have negative effects on human
health (Romano & Suzzi, 1993). Therefore, wine industry is chal-
lenged to meet consumers' demands of reducing the levels of SO2
used in wine production (du Toit & Pretorius, 2000). Techniques
such as ltration and ning are efcient in controlling microbial growth,
but unfortunately these techniques have detrimental effects on the sen-
sorial properties of the wine (Armada & Falqu, 2007; Bartowsky, 2009;
Fredericks, du Toit, & Krgel, 2011; Gergely, Bekassy-Molnar, & Vatai,
2003; Lpez, Castro, Garca, Pazo, & Barroso, 2001; Surez, Surez-
Lepe, Morata, & Caldern, 2007).
In modern times, the quality and safety of food products are
among the most important factors that inuence the consumer
choices (Considine, Kelly, Fitzgerald, Hill, & Sleator, 2008). Today's
consumers demand for high quality foods that are free from additives,
fresh tasting, microbiologically safe and with an extended shelf-life.
This led the food industry to investigate alternative processing
methods (Balasubramaniam, Ting, Stewart, & Robbins, 2004).
Among these methods high hydrostatic pressure (HHP) is, perhaps,
the most popular one. This nonthermal processing technique is an ef-
fective method for increasing food safety and shelf life while preserv-
ing the organoleptic properties of food products (Farkas & Hoover,
2000; San Martn, Barbosa-Cnovas, & Swanson, 2002).
During the last 20 years scientic studies on the effect of HHP on var-
ious food products has been explored (Alemn et al., 1996; Buzrul, Alpas,
Largeteau, Bozoglu, & Demazeau, 2008; Buzrul, Alpas, Largeteau, &
Demazeau, 2008; Chen & Hoover, 2004; Del Pozo-Insfran, Del
Follo-Martinez, Talcott, & Brenes, 2007; El Moueffak et al., 1995;
Garcia-Graells, Hauben, & Michiels, 1998; Gervilla, Capellas, Ferragut,
& Guamis, 1997; Goodridge, Willford, & Kalchayanand, 2006;
Lpez-Caballero, Prez-Mateos, Montero, & Borderas, 2000;
Ogawa, Fukuhisa, Kubo, & Fukumoto, 1990; Rasanayagam et al.,
2003; Shao, Ramaswamy, & Zhu, 2007; Tanaka & Hatanaka, 1992;
Van Opstal, Vanmuysen, Wuytack, Masschalck, & Michiels, 2005;
Yuste, Pla, Capellas, & Mor-Mur, 2002). Currently, a whole range of
food products such as fruit juices, sea foods and meat products can be
found on market shelves all around the world (Matser, Krebbers, van
den Berg, & Bartels, 2004). However, few data have been reported about
the use of HHP on beer and wine. Although, rice wine (nigori-sake) is
one of the earliest HHP-treated commercial products that appeared on
the Japanese market (Suzuki, 2002), no HHP-treated alcoholic beverage
such as beer and wine is introduced in the market throughout the
world. The objective of this contribution is to compile the studies of
HHP on beer and wine. The microbiological, biochemical and sensorial as-
pects of HHP-treated beer and wine will also be underlined.
2. Applications of HHP on brewing process and beer
To the best of author's knowledge, rst trial of HHP on brewing pro-
cess and beer was done by Fischer, Schberl, Russ, and Meyer-Pittroff
(1998). Fischer et al. (1998) applied HHP on mash, wort and beer. For
the treatment of mash, 50 g of malt grain with 100 mL of distilled
water in polyethylene bags at room temperature (RT) was pressurized
at 300, 500 and 700 MPa for 5 min. Untreated mash sample was used
as the control. The content of solved protein, high-molecular and low-
molecular nitrogen of HHP-treated mash samples increased as the pres-
sure increased, compared to untreated mash sample. The coagulatable
nitrogen increased up to 500 MPa and dropped thereafter. Free amino
nitrogen (FAN) decreased compared to control. Viscosity decreased as
the pressure increased. At 300 MPa saccharication slightly took place
but at higher pressures it was not observed. Fermentation degree
dropped with the increasing pressure. No changes were determined
for pH value.
For the treatment of wort by Fischer et al. (1998), wort samples
were pressurized at 300, 500 and 700 MPa for 5 min, and additionally
a sample was treated at 700 MPa for 30 min. As the controls wort
samples were cooked for 30 and 60 min at atmospheric pressure
(0.1 MPa). The results revealed that the bitterness and the amount
of iso--acids could be increased more by conventional cooking
than HHP treatment. Among the HHP-treated samples best isomeri-
zation was obtained at 700 MPa for 30 min.
Fischer et al. (1998) tried 300, 500 and 700 MPa for 5 min treat-
ments on bright lager beer samples in polyethylene naphthalate
(PEN) bottles. Comparison was done with untreated and pasteurized
(60 C for 20 min) beer samples. HHP treatment did not signicantly
change the color, foam durability and the spectrum of avor mate-
rials. However, turbidity increased at 500 and 700 MPa. The potential
for the arising of turbidity dropped by the HHP treatment compared
to untreated and heat pasteurized beer samples.
Castellari, Arfelli, Riponi, Carpi, and Amati (2000) compared HHP
and heat pasteurization of beer. Two pale ales and mild ale beers
were used. First trial was made with a pale ale beer. Unltered beer
samples were taken directly from the storage tanks at the maturation
phase. Samples were treated at 600 MPa for 5 min, and heat treat-
ment was made at 60 C for 10 min. Untreated sample was used as
control. Results revealed that both HHP and heat treatments did not
affect the main chemical constituents of the beer (insignicant differ-
ences for pH, ethanol, extract, bitterness, iso--acids, catechins were
obtained for both treatments compared to control). On the contrary,
heat treatment increased the color parameters (L, a, b), while
HHP-treated samples did not signicantly differ from color of
untreated beer. Heat pasteurization signicantly reduced permanent
haze compared to untreated and HHP samples. Both HHP and heat
treatments reduced total aerobic, yeast and molds counts about 4
log10 cycles. No lactic acid bacteria were detected in either of the sta-
bilized beer samples.
In a second trial a pale ale and a mild ale were used by Castellari
et al. (2000). Samples were treated by HHP (600 MPa for 5 min)
and heat (60 C for 10 min). All samples were then stored in the
dark at 20 C. At 1, 8, 14, 26 and 49 days of storage one bottle from
each of the treatments was analyzed. HHP and the heat pasteurization
affected the pH, bitterness and phenols content of beers up to 49 days
of storage similarly. Under these conditions, the storage time had no
signicant inuence on these parameters (except for bitterness) and
no interactions between factors were found for these analytical pa-
rameters. Hydroxymethylfurfural (HMF) increased signicantly with
heat pasteurization. The HHP beers retained a signicantly higher
permanent haze throughout the storage period. The heat pasteurized
beers showed a sharp decrease of Nephelos Turbidity Unit (NTU)
values in the rst days of storage, with an increase of chill haze values
at the same time. Permanent haze was more inuenced by the stabi-
lizing process in pale ale than in mild ale.
The different stabilization processes had no inuence on the light-
ness (L), but all the other color indices were higher in heat processed
samples. During storage the chromatic index a (red-green) of HHP
treated pale ale increased more slowly while the b (yellow-blue)
value decreased in a more dramatic way than in the corresponding
heat treated samples. As a result of combining effects of these trends,
the C (color saturation) decreased in all samples during storage and
was always lower in HHP beers.
The microbiological analyses conrmed that both HHP and heat
treatments gave comparable microbial inactivation. No signicant in-
creases in microbiological counts and no lactic acid bacteria develop-
ment were observed during the entire storage period (49 days) at
20 C for both heat pasteurized and HHP samples.
Ulmer, Gnzle, and Vogel (2000) investigated the effects of HHP
on survival, metabolic activity, membrane integrity and hop resis-
tance of Lactobacillus plantarum TMW1.460 [a highly hop-resistant
(possesses a plasmid-encoded hop resistant mechanism, HorA) or-
ganism isolated from spoiled beer] in model beer (MB). MB was pre-
pared by inoculating malt extract medium with Saccharomyces
cerevisiae to a cell count of about 5 10 6 cells mL 1. The mash was
 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112
fermented for 140 h at 10 C, heat sterilized and the yeast was re-
moved by centrifugation. The clear supernatant was collected, residu-
al ethanol and carbon dioxide (CO2) were removed in a rotary
evaporator under vacuum, and the weight loss was compensated for
with demineralized water. The pH was adjusted to 4.0, and the medi-
um was sterilized at 121 C for 21 min. Inoculated cells of L. plantarum
in MB were HHP treated at 200, 400, 500, 600 MPa and 15 C for var-
ious holding times.
Inoculation of HHP-treated L. plantarum in MB on non-selective
(MRS) and selective (MRS-NaCl) media demonstrated sublethal inju-
ry during HHP treatment; however at longer holding times (more
than 60, 12 or 2 min at 200, 400 or 500 MPa, respectively), sublethal
injury was no longer observed. Sigmoidal (starting with downward
concavity and ending up with upward concavity) survival curve of
HHP-treated L. plantarum in MB was observed at 200 MPa, whereas
only tailing (upward concavity) was observed for survival curves at
400 and 500 MPa. At 600 MPa, all the cells were killed during com-
pression period.
Diffusion of ethidium bromide (EB) through membranes of pres-
surized cells of L. plantarum in MB indicated that membrane damage
occurred prior to cell death. Pressurization with 600 MPa resulted in
complete loss of metabolic activity and membrane integrity. Cells
pressurized at 200 MPa for 0 to 12 min were able to maintain an in-
ternal low EB concentration although EB diffusion across the mem-
brane was facilitated. During the rst 12 min, no loss of cell viability
or sublethal injury was observed by plate counts. After a 30- and a
60-min pressure holding time, the cultures lost their ability to ex-
clude EB, indicating loss of hop resistance.
Gnzle, Ulmer, and Vogel (2001) investigated the effect of ethanol
and hop extract on inactivation of L. plantarum in MB system during
and after HHP treatment. MB was prepared as described by Ulmer
et al. (2000). To obtain MB with specied content of ethanol and
hop extract, ethanol [5 and 10% (w/w)] and isomerized hop extract
containing 22% iso--acids (50 and 100 ppm) were used. All experi-
ments were performed at 300 MPa and 20 C for various holding
times. Prior to HHP experiments cells were inoculated into MB, MB
containing 5 and 10% ethanol or MB containing 50 and 100 ppm
hop extract. Non-linear survival curves were observed. Addition of
ethanol and hop extract accelerated HHP inactivation of L. plantarum
in MB. In the absence of additives about 10 min was required to re-
duce the viable cell counts by 99% and uninjured cells by 99.99%
whereas 6 min was enough in the presence of 100 ppm hop extract
to obtain the same effect. Ethanol added at the level of 10% had a
much stronger effect on pressure inactivation than 100 ppm hop
concentration.
To observe the survival of HHP-treated L. plantarum, Gnzle et al.
(2001) employed 300 MPa 5 min treatment. Survival was monitored
in MB (no addition of either hops or ethanol), MB containing
50 ppm hop extract, 5% ethanol, or both. Additionally, commercially
available beer was used for comparison. HHP treatment reduced the
viable cell counts by 90% and rendered 90% of the surviving cells sub-
lethally injured. A reduction of viable cell counts was observed during
storage (storage temperature was 10 C) of HHP-treated cells in MB
and MB containing 5% ethanol. Sublethally injured cells were killed
within 48 h whereas uninjured cells remained unaffected by cold
storage. An appreciable effect of ethanol on either viable or sublethal
injury was not observed. The presence of hop compounds during
storage had a far pronounced effect on survival of HHP-treated
L. plantarum than ethanol. Hop extract in a concentration of 50 ppm
resulted in sublethal injury of virtually all cells within 3 h of storage.
Viable cell counts were reduced by 90% within 3 h, indicating rapid
inactivation of cells that were already sublethally injured by HHP
treatment, and viable cell counts dropped below the detection level
after 27 h of storage. In commercially available beer, cell counts
were below the detection level on either non-selective (MRS) or se-
lective (MRS-NaCl) agar after 2 h of storage.
3
Prez-Lamela, Ledward, Reed, and Simal-Gndara (2001) applied
HHP to accelerate the process of malting: 5 g of barley with 5 mL of
distilled water in plastic bags was pressurized at 40, 60, 100, 200
and 400 MPa for 20 min. Untreated sample steeped in water for 8 h
at 0.1 MPa used as the control. Results showed that increase in
water from 12% to 20% could be achieved by HHP treatment and the
water distribution increased from 62% to 84%. Treatment of
400 MPa for 20 min had the similar values with that of control.
To determine the viability of pressurized barley grains, samples were
treated at 25, 50, 100, 400 and 600 MPa for 20 min by Prez-Lamela et al.
(2001). Untreated sample steeped in water for 8 h used as the control.
Hydrogen peroxide test and staining method indicated that viability of
the grains was practically unaffected by the application of HHP.
To determine the cold water extract (CWE) [CWE values are indica-
tive of over-modication of the malt with associated loss of product,
i.e., lower hot water extract (HWE) values, as a result of excessive enzy-
matic activity] of pressurized germinated barley (green malt), samples
were treated at 50, 100, 200 and 400 MPa for 20 min by Prez-Lamela
et al. (2001). Untreated sample steeped in water for 8 h used as the con-
trol. Green malt samples treated with HHP had higher CWE values than
the control; however, HWE values were not affected by HHP indicating
that a loss of product did not occur.
Prez-Lamela, Ledward, Reed, and Simal-Gndara (2002) applied
HHP at RT to the mashing of white malt. For the -glucanase (this en-
zyme breaks down gums and -glucans in wort and is desirable to ob-
tain good quality beer) activity measurement batches of 20 g of malt
were milled. Batches of 5 g of milled malt were weighed into plastic
bags, 46 mL of distilled water was added to each and the samples
were pressurized at 200, 300, 400, 500 and 600 MPa for 20 min. A
control of 5 g of milled malt was left at 0.1 MPa with 46 mL of distilled
water for 20 min. The -glucanase activity in wet-treated malt
showed a decrease compared to control samples following HHP. No
signicant difference was observed between different pressure
values.
For starch gelatinization measurement by Prez-Lamela et al. (2002)
mashing liquid obtained by mixing 5 g of milled malt and 46 mL of dis-
tilled water at RT was sealed in separate plastic bags. One sample was
left at 0.1 MPa for 20 min (control) and the others were treated at 400
and 600 MPa for 20 min. HHP caused the gelatinization of starch: dis-
ruption became more marked at 600 MPa than 400 MPa.
Sugar (sucrose in g per 100 mL) and extract (l kg 1) values were
also determined by Prez-Lamela et al. (2002): batches of 5 g of
milled malt and 46 mL of distilled water were sealed in plastic bags
at RT. One sample was left at 0.1 MPa for 3 h (untreated control),
one sample was treated at 65 C for 35 min to mimic the mashing
process in the brew-house (heat treated control) and the others
were treated at 50, 100, 200, 400 and 600 MPa for 20 min, respective-
ly. At higher pressures (400 and 600 MPa), sucrose content was dou-
bled compared with that found at lower pressures (50200 MPa).
Although sucrose and extract values of HHP treated samples were
lower than the heat treated control (65 C for 35 min), 400 and
600 MPa treatments resulted in much higher values (of sucrose and
extract values) than the untreated control.
Ulmer, Gnzle, and Vogel (2002) investigated the HHP inactiva-
tion of L. plantarum and S. uvarum in MB. MB was prepared as de-
scribed by Ulmer et al. (2000). Inoculated cells of L. plantarum in
MB were HHP treated at 200, 300, 400, 500, 600 MPa and 15 C for
various holding times. Similar results with that of Ulmer et al.
(2000) were obtained: As the pressure increased inactivation in-
creased. At 200 MPa more than 60 min was required for about 6
log10 reduction while at 600 MPa compression period resulted in
more than 6 log10 reduction. HHP treatment of MB with hop extract
(50 and 100 mg L 1) and ethanol (5 and 10%) revealed that addition
of ethanol and hop extract accelerated HHP (300 MPa for various
times at 15 C) inactivation of L. plantarum in MB. Addition of ethanol
had stronger effect on pressure inactivation than the addition of hop
4 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112
extract. HHP treatment (300 MPa for various times at 15 C) also en-
hanced the loss of metabolic and HorA activities of L. plantarum in MB.
Effect of pH value for HHP (500 MPa for 5 min at 15 C) inactivation
on L. plantarum in MB was also studied by Ulmer et al. (2002). No inacti-
vation but only sublethal injury was observed at pH 6 and 7 whereas
more than 5 log10 inactivation took place at pH values lower than 5.
Storage (at 10 C) of MB with 50 mg L 1 hop extract and 0% etha-
nol and MB with 50 mg L 1 hop extract and 5% ethanol after HHP
treatment (300 MPa for 5 min at 15 C) indicated that L. plantarum
cells died completely within 24 h.
HHP treatment indicated that L. plantarum was more pressure tol-
erant than S. uvarum. However, storage (at 15 C) of MB with
50 mg L 1 hop extract after HHP (100 MPa and 200 MPa for 5 and
10 min at 15 C) showed that number of cells of S. uvarum in MB
remained stable while HHP-treated L. plantarum died within 32 h.
Fischer, Russ, and Meyer-Pittroff (2002) treated the wort and beer
by HHP. Wort samples were pressurized at 300, 500 and 700 MPa for
5 min, and additionally a sample was treated at 700 MPa for 30 min.
As the controls wort samples were cooked for 30 and 60 min at atmo-
spheric pressure just like Fischer et al. (1998) did before. Similar re-
sults with that of Fischer et al. (1998) were obtained: clear increase
of iso--acids was detected at 700 MPa. Longer treatment time
(30 min instead of 5 min) led to even better results. HHP treatment
reduced the treatment time because there were no heating and cool-
ing time by this treatment. An extra treatment at 600 MPa for 30 min
was employed to identify iso--acids using HPLC. A signicant HHP
induced isomerization occurred, but only one peak was in the area
of thermal isomerization products. The other HHP induced isomeriza-
tion products fullled the characteristics of iso--acids, but they were
not identical with the thermal ones.
Two different types of beer were used by Fischer et al. (2002): one
with no ltration problems, the other with enormous ltration prob-
lems. HHP treatments were performed at 300, 500 and 700 MPa for
5 min. Trials performed with the shift lters indicated that lterabili-
ty of the beer with no ltration problems became better: ltration
time decreased to 50%. The sample treated at 300 MPa was even com-
parable with water. Similar results were obtained for the beer with
enormous ltration problems: ltration time decreased to about
50% at 300 MPa, and the level of ltration time was a bit higher
than the other beer. The effect of HHP on lterability of beer on kie-
selguhr lter revealed the improvement of the specic lterability
as the shift lter. The only difference was that the best effect had
been achieved at 500 MPa. To understand the effect of HHP on the im-
provement of beer lterability several substances responsible for the
lterability were determined: polyphenols, anthocyanogens, protein
fractions and particle size showed no change. Only the content of -
glucan-gel decreased. At 300 and 500 MPa -glucan-gel content
could not be detected.
Two types of beer (Lager and Pilsener) were used by Fischer et al.
(2002) to determine the effects of HHP on colloidal stability. No
change was observed for the turbidity of both beers at 300 and
500 MPa compared to untreated samples. Potential for arising the tur-
bidity decreased at 300 and 500 MPa and increased at 700 MPa indi-
cating that stability (up to 500 MPa) became better. Pilsener type
beer was more stable because of the higher content of hop
ingredients.
Another study done on the effects of HHP on wort and beer was
that of Prez-Lamela, Reed, and Simal-Gndara (2004). Two types of
wort were pressurized: 11 and 16 (% sugars) Plato lager worts.
11 Plato wort samples were poured into plastic bags, sealed and
treated at 200, 400 and 600 MPa for 20 min. To investigate the inu-
ence of time, 16 Plato wort samples were treated at 600 MPa for 5, 10
and 20 min. Untreated samples at 0.1 MPa and heat treated samples
(65 C for 90 min) at 0.1 MPa were used as controls. Results of 11
Plato wort samples treated by HHP indicated that HHP did not affect
pH. Heat treatment increased the color while the color of HHP-
treated and untreated samples were approximately 25% lower than
heat treated wort. Dimethyl sulde (DMS) decreased by the applica-
tion of heat whereas untreated and HHP-treated samples had the
similar values for DMS. Bitterness value for heat treated sample was
much higher than the ones for untreated and HHP-treated samples.
Similar results were obtained for the treatment of 16 Plato wort sam-
ples: pH did not change, bitterness of heat treated wort sample was
higher. Moreover, iso--acids and total -acids of the heat treated
sample were also higher. Holding time of HHP treatment on 16
Plato wort samples had no effect on pH, bitterness, iso--acids, total
-acids, total protein and -glucan.
Lager and ale beers were used by Prez-Lamela et al. (2004). Sam-
ples were poured into plastic bags and treated at 300, 400, 500 and
600 MPa for 20 min. Untreated samples at 0.1 MPa were used as con-
trols. HHP treatment of beer resulted in an increase in the foaming
and haze characteristics of the beer. Product stability, directly related
to saturated ammonium sulfate precipitation limit (SASPL) value, was
improved with pressurization, especially at 500600 MPa.
Buckow, Heinz, and Knorr (2005) investigated the combined ef-
fect of HHP (0.1900 MPa) and temperature (3075 C) on the activ-
ity of -glucanase from barley malt. Highest thermostability of -
glucanase was found at 400 MPa. At temperatures above 55 C and
ambient pressure, as well as at 900 MPa and 40 C, high inactivation
of -glucanase was observed resulting in a complete loss of enzyme
activity in less than 30 min. Enzymatic catalysis was found to be
delayed by pressures up to 600 MPa. For the overall depolymerization
reaction of -glucans in ACES buffer (0.1 M, pH 5.6) a maximum was
identied at 215 MPa and 55 C yielding approximately 2/3 higher
degradation of -glucan after 20 min as compared to the maximum
at ambient pressure (45 C). Authors concluded that although the ef-
fect of pressure and temperature on the catalytic potential of -
glucanase was investigated in a model system (0.1 M ACES buffer,
pH 5.6), the results could be transferred to the real mashing process
with possible small deviations in enzyme stability due to the protect-
ing effects of high solid and ion concentrations. The reported effect
might be suitable for other mashing enzymes as well.
Effect of HHP on quality parameters of lager beer was studied by
Buzrul, Alpas, and Bozoglu (2005a). Unpasteurized lager beer samples
from a commercial brewery were treated either by HHP (200, 250,
300, 350 MPa for 3 and 5 min at 20 C) or by conventional heat pas-
teurization (60 C for 15 min). The main attributes of the beer, such
as ethanol content, density, extract, fermentation degree and pH,
were not affected by either treatment; however, HHP and heat pas-
teurization affected color, chill haze, protein sensitivity and bitter-
ness. The color, protein sensitivity and chill haze values increased as
the pressure and pressurization time increased. Change in bitterness
was higher in conventional heat pasteurization, but pressures up to
300 MPa had no signicant affect on bitterness.
In another study Buzrul et al. (2005a) investigated the effects of
HHP on shelf-life of lager beer. Filtered bright lager beer samples
were either treated with HHP (350 MPa for 3 and 5 min at 20 C) or
conventional heat pasteurization (60 C for 15 min). A storage period
of 56 days showed that HHP and heat pasteurization had similar re-
sults in terms of pH and color. However, HHP-treated samples had
lower bitterness and protein sensitivity and higher chill haze values
than the heat pasteurized samples at the end of the storage period.
The microbiological stability of HHP-treated beers was the same as
that of heat-treated beers, and the development of both lactic and
acetic acid bacteria was inhibited for 56 days of storage.
Effects of HHP on microbiological and technological characteristics
of beer were studied by Fischer et al. (2006): -amylase from barley
in ACES was HHP treated at 0.1700 MPa, 2070 C for various hold-
ing times. Results showed that temperature tolerance increased
with rising pressure. Inactivation increased at 0.1 MPa and above
500 MPa. Moreover, at 200 and 250 MPa (even at 65 C) inactivation
rate was not very signicant. -amylase was most stable at 200 MPa.
 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112
Amyloglucosidase in ACES was HHP treated at 0.11200 MPa,
2090 C for various holding times. Glucose produced from maltose
monohydrate indicated that the optimum could be reached at
370 MPa and 85 C which was about 35% higher in comparison to
the optimum at 0.1 MPa and 65 C. The best temperature tolerance
was at about 700 MPa. To reach an inactivation of 90%, temperatures
of approximately 105 C were necessary. -glucanase from Bacillus
subtilis in ACES buffer were HHP treated at 0.11000 MPa, 3070 C
for various holding times. A signicant stabilization against thermal
inuence at 400 MPa and an increase of the transferring rate at
200 MPa and 58 C of about 40% were measured.
The examinations of the lterability and behavior of -glucan gel
were carried out with beer and model systems by Fischer et al.
(2006). The model systems were produced of isolated -glucan
from barley. In a rst attempt, unltered commercial beer with ltra-
tion problems was HHP treated at 300, 500, 700 MPa, 20 C for 5 min.
The analysis was carried out with a cellulose shift lter. Secondly, the
beer samples were HHP treated at 100, 200, 300, 400 and 500 MPa,
20 C for 1, 150, 300 and 500 s. The analysis was carried out with kie-
selguhr lter. It was observed that the lterability increased with ris-
ing pressure: the optimum was reached between 300 and 500 MPa.
The reason for this improvement was the decrease in the -glucan
gel content: As the pressure increased -glucan gel content decreased
in both beer and model system. HHP treatment (especially at and
above 500 MPa) resulted in a decrease in viscosity of the samples
with different -glucan gel concentrations.
Fischer et al. (2006) examined the effect of pH (4.0, 5.0, 6.0 and
6.5) and HHP treatment (300 MPa, 20 C for various holding times
up to 120 min) on L. plantarum (moderately hop tolerant) in MB. It
was found that inactivation was more effective at lower pH values:
5 log10 reduction could be achieved. Sublethal injury was observed
at short pressurization times. HHP treatment (300 MPa, 20 C for
60 min) and following storage at a pH value of 6.5 affected only a deg-
radation of 20% of the content of colony forming units. Storage at pH
5.0 and 4.0 resulted in a reduction of 90% to 95%.
HHP treatment of L. plantarum at 200 MPa and neutral pH for var-
ious holding times (up to 32 min) showed insignicant reduction;
however, in model beer with pH 4.0 the membrane transport system
HorA was completely inactivated rendering the cells hop sensitive.
Consequently, the cells with lowered hop resistance died in the pres-
ence of hops within a few hours. HHP treatment (200 MPa up to
32 min) of L. brevis in beers with low (16.7 mg L 1 isohumolone)
and high hop content (35.8 mg L 1 isohumolone) at 200 MPa for var-
ious holding times (up to 32 min) also showed no signicant reduc-
tion; however, upon storage inactivation was not observed
indicating this strain was highly hop tolerant.
Table 1 summarizes the studies done about brewing process and beer.
3. Applications of HHP on wine
First report about HHP on wine was that of Lonvaud-Funel,
Dupont, Demazeau, and Bignon (1994). Sauternes (a sweet white
wine from Bordeaux, France) wine (pH 3.93, 12.3% ethanol, 70 g L 1
reducing sugars) was treated with HHP (200400 MPa) for various
holding times. Initial yeast count (6.88 log10) decreased to 4.88 and
4.54 log10 after HHP treatment at 200 and 250 MPa, respectively,
whereas more than 3 log10 reduction was possible after pressuriza-
tion at 250 MPa for 20 min. Pressurization at 370 and 400 MPa for
5 min, 300 MPa for 10 and 20 min, 350 and 400 MPa for 15 min
resulted in complete elimination of yeasts. Three days after (samples
were kept at 25 C during 3 days) the HHP treatment, samples were
again treated at 250 and 300 MPa for 10 and 20 min. The results
showed that yeast counts were 4.95 and 2.48 log10 after pressuriza-
tion at 250 MPa for 10 and 20 min, respectively, whereas no colonies
were observed for 10 and 20 min treatment at 300 MPa.
5
Sauternes wine treated at 300 MPa for various times indicated
that initial yeast count (6.30 log10) decreased to 4.18 and 3.15 log10
after HHP treatment for 1 and 5 min, respectively, whereas complete
elimination of yeasts was possible for 10 and 15 min. Inactivation fur-
ther increased after 3 days of storage. Viscosities of the wine samples
treated at 300 MPa for 1, 5, 10 and 15 min were similar to that of
untreated wine. The optical density (color values) of untreated wine
was higher than that of HHP treated wine samples (300 MPa for 1,
5, 10 and 15 min). Due to oxidation during storage color values in-
creased; however, untreated wine was darker than the HHP-treated
wines.
Sensory evaluation was done by 5 specialists on three wines: a)
Sulte-added wine (250 mg L 1) b) HHP-treated (350 MPa for
10 min) and sulte-added wine (250 mg L 1) c) HHP-treated
(350 MPa for 10 min) wine. Tasting of the wines was done 12 days
after the treatments. First evaluation was done between (a) and (b),
and the second between (a) and (c). In the rst evaluation none of
the tasters succeeded in differentiating the wines whereas in the sec-
ond evaluation HHP-treated wine was easily recognized by the tasters
because of oxidation.
Second study about HHP on wine was done by Delni et al.
(1995). Barbera grape must (pH 3.0, sugars 177.2 g L 1, no SO2 and
ethanol present) and Moscato wines obtained from the same grape
must were used. Two Moscato wines obtained had the following
compositions a) 8.22% ethanol, 64 mg L 1 total SO2, 146 mg L 1 acet-
aldehyde, 50.5 g L 1 residual sugars; b) 11.35% ethanol, 102.4 mg L 1
total SO2, 168 mg L 1 acetaldehyde, no residual sugars; a third wine
(c) was obtained by adding absolute ethanol to (b) up to 15.19%.
The SO2 was produced by yeast during fermentation. Mixture of
yeast, molds, lactic and acetic acid bacteria were used. HHP treat-
ments (300 to 600 MPa) were done at 20 C initially.
The results showed that HHP (600 MPa for 3 min) had a strong an-
timicrobial effect: 6 log10 (complete inactivation) was observed for all
types of wine (a, b and c). At 400 MPa for 2 min all the microorgan-
isms added to the sweet wine (a) died, while at 300 MPa for 2 min
and 4 min the survival rates were 19.67 and 10.57%, respectively.
Treatment (600 MPa for 2 min) of Asti Spumante wine (pH 3.4,
64 mg L 1 SO2) resulted in no relevant variation of the color param-
eters compared with the untreated wine, except a weak browning
tendency over time (90 days of storage) that can be considered insig-
nicant for the very low SO2 content. The oxygen concentration exist-
ing in the samples after 6 h from HHP treatment was signicantly
higher in the untreated samples than in the pressurized ones; this ob-
servation however, was not conrmed after 24 h. In addition, the de-
crease of dissolved oxygen content appeared stronger in the treated
samples, but only in the rst 5 min after air injection.
Tonello, Largeteau, Demazeau, and Lonvaud-Funel (1996a) used
HHP treatment to inactivate yeasts, lactic and acetic acid bacteria in
different wines (white, red and ros). Alcohol was used to adjust eth-
anol levels at 9, 11, 13 and 15%. Treatment of 300 MPa for 6 min at RT
resulted in complete elimination of yeasts in all wines; however
when the same treatment was applied for only 1 min signicant in-
crease in inactivation of yeasts was observed for the wines containing
13 and 15% ethanol compared to wines containing 9 and 11% ethanol.
There was no difference in inactivation of yeasts when sugar (8 g L 1
to 98 g L 1) was added to wines.
Two types of liqueur-like white wine (one had 3.2 log10 the other
had 0.4 log10 yeasts) and two types of red wine (one had 3.1 log10 the
other had 2.9 log10 yeasts) were treated at 400 MPa for 20 s, 1 and
2 min at RT. All treatments were effective for the reduction of yeasts
in wines: treatment at 400 MPa for 2 min completely eliminated the
yeasts in liqueur-like white wine containing 0.4 log10 yeasts and red
wine containing 3.4 log10 yeasts whereas only small numbers of
yeasts managed to survive in other white and red wines.
Same red wines with lactic and acetic acid bacteria (one had 3 log10
lactic acid and 5.2 log10 acetic acid bacteria the other had 2.2 log10 lactic
6 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112

Table 1
Applications of high hydrostatic pressure (HHP) on brewing process and beer.

Pressure, temperature, time Sample treated Achievements Reference

40, 60, 100, 200 and 400 MPa, 20 min 5 g of barley with 5 mL of distilled water Increase in water and Prez-
Lamela
water distribution.
et al.
(2001)
100, 400 and 600 MPa, 20 min 5 g of barley with 5 mL of distilled water Viability of the grains Prez-
Lamela
was unaffected.
et al.
(2001)
50, 100, 200 and 400 MPa, 20 min Germinated barley Higher cold water ex- Prez-
Lamela
tract values.
et al.
No effect for hot water (2001)
extract value.
200, 300, 400, 500 and 600 MPa, 20 min 5 g of milled malt with 46 mL of distilled water Decrease of - Prez-
Lamela
glucanase activity in
et al.
wet-treated malt. (2002)
No difference between
different pressure
values.
400 and 600 MPa, 20 min 5 g of milled malt with 46 mL of distilled water HHP caused the gela- Prez-
Lamela
tinization of starch.
et al.
(2002)
50, 100, 200, 400 and 600 MPa, 20 min 5 g of milled malt with 46 mL of distilled water Sucrose content was Prez-
Lamela
higher at higher
et al.
pressures. (2002)
Sucrose and extract
values were lower
than the heat treated
samples, higher than
the untreated control.
300,500 and 700 MPa, room temperature, 5 min Mash Solved protein, high- Fischer
et al.
molecular, low-
(1998)
molecular and coagu-
latable nitrogen
increased.
Free amino nitrogen,
viscosity and fermen-
tation degree
decreased.
Saccharication
slightly took place.
No changes for pH.
0.1900 MPa, 3075 C -glucanase from barley malt in ACES buffer Increase in Buckow
et al.
thermostability.
(2005)
High inactivation at
>55 C and 0.1 MPa,
and at 900 MPa and
40 C.
0.1700 MPa, 2070 C, various holding times -amylase from barley in ACES buffer Temperature tolerance Fischer
et al.
increased.
(2006)
Inactivation increased
at 0.1 MPa and above
500 MPa.
0.11200 MPa, 2090 C, various holding times Amyloglucosidase in ACES buffer The best temperature Fischer
et al.
tolerance was at about
(2006)
700 MPa.
To reach an inactivation
of 90%, temperatures of
approximately 105 C
were necessary.
0.11000 MPa, 3070 C, various holding times -glucanase from Bacillus subtilis in ACES buffer Stabilization against Fischer
et al.
thermal inuence at
(2006)
400 MPa.
 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112 7

Table 1 (continued)
Pressure, temperature, time Sample treated Achievements Reference

Increase of the trans-

ferring rate.
300, 500 and 700 MPa, 5 min Wort Bitterness and the Fischer
+ et al.
amount of iso--acids
700 MPa, 30 min (1998)
increased more by
heat than HHP.
Best isomerization was
obtained at 700 MPa for
30 min.
300, 500 and 700 MPa, 5 min Wort Similar results with Fischer
+ et al.
that of Fischer et al.
600 and 700 MPa, 30 min (2002)
(1998).
200, 400 and 600 MPa, 20 min 11 Plato wort No effect on pH. Prez-
Lamela
Lower color values of
et al.
than heated wort. (2004)
HHP-treated samples
had similar values for
dimethyl sulde.
Bitterness for heat trea-
ted sample was higher.
600 MPa, 5, 10 and 20 min 16 Plato wort pH did not change, Prez-
Lamela et
bitterness of heat trea-
al. (2004)
ted wort sample was
higher.
No effect of holding
time on pH, bitterness,
iso--acids, total -
acids, total protein and
-Glucan.
300, 500 and 700 MPa, 5 min Beer No change for color, Fischer
et al.
foam durability and
(1998)
the spectrum of avor
materials.
Increase in turbidity.
Decrease in potential
for the arising of
turbidity.
600 MPa, 5 min Unltered and unpasteurized pale ale beer No effect on main Castellari
et al.
chemical constituents
(2000)
of the beer.
No signicant changes
on color.
Reduction of total aero-
bic, yeast and molds
counts.
No lactic acid bacteria
were detected.
600 MPa , 5 min Unltered and unpasteurized pale ale and mild ale beers HHP and heat affected Castellari
et al.
the pH, bitterness and
(2000)
phenols content of
beers up to 49 days of
storage similarly.
High permanent haze
throughout the storage
period.
Permanent haze was
more inuenced in pale
ale than in mild ale.
No increase in microbi-
ological counts and no
lactic acid bacteria de-
velopment were
(continued on next page)
8 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112

Table 1 (continued)
Pressure, temperature, time Sample treated Achievements Reference

observed during the

entire storage period.
300, 500 and 700 MPa, 5 min Beer with no ltration problems Filterability became Fischer
+ et al.
better: ltration time
Beer with enormous ltration problems (2002)
decreased to 50%.
For the beer with
enormous ltration
problems: the level of
ltration time was
higher than the other
beer.
300, 500 and 700 MPa, 5 min Lager and Pilsener beers No change was ob- Fischer
et al.
served for the turbidity.
(2002)
Potential for arising the
turbidity decreased at
300 and 500 MPa and
increased at 700 MPa.
Pilsener type beer was
more stable.
300, 400, 500 and 600 MPa, 20 min Lager and ale beers Increase in the foam- Prez-
Lamela
ing and haze charac-
et al.
teristics of the beer. (2004)
Improvement of satu-
rated ammonium sul-
fate precipitation limit
value.
200, 250, 300 and 350 MPa, 20 C, 3 and 5 min Unpasteurized lager beer Main attributes of the Buzrul
et al.
beer were not affected.
(2005a)
The color, protein
sensitivity and chill
haze increased as the
pressure and time
increased.
No effect on bitterness
up to 300 MPa.
350 MPa, 20 C, 3 and 5 min Unpasteurized lager beer Similar results in Buzrul,
et al.
terms of pH and color
(2005b)
for HHP and heat dur-
ing 56 days.
HHP-treated samples
had lower bitterness
and protein sensitivity
and higher chill haze
values than the heat
pasteurized samples.
Development of lactic
and acetic acid bacte-
ria was inhibited.
300, 500 and 700 MPa, 20 C, 5 min Unltered commercial beer with ltration problems Filterability increased Fischer
+ et al.
with rising pressure.
100, 200, 300, 400 and 500 MPa, 20 C, 1, 150, 300 and 500 s (2006)
200 MPa, various holding times up to 32 min Low hop content beer with L. plantarum or L. brevis+ L. brevis was highly Fischer
High hop content beer with L. plantarum or L. brevis et al.
hop tolerant.
(2006)
200, 400, 500 and 600 MPa, 15 C, various holding times Model beer (MB) with Lactobacillus plantarum Sublethal injury and Ulmer
et al.
non-linear survival
(2000)
curves were observed.
Membrane damage
occurred prior to cell
death.
Complete loss of met-
abolic activity and
membrane integrity.
Loss of hop resistance.
 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112 9

Table 1 (continued)
Pressure, temperature, time Sample treated Achievements Reference

300 MPa, 20 C, various holding times MB with L. plantarum Ethanol and hop ex- Gnzle
et al.
tract accelerated HHP
(2001)
inactivation.
Ethanol had much
stronger effect on in-
activation than hop
extract.
300 MPa, 20 C, 5 min MB with L. plantarum A reduction of cells Gnzle
+ et al.
was observed during
Commercial beer with L. plantarum (2001)
storage (at 10 C).
Sublethally injured
cells were killed with-
in 48 h.
The presence of hop
compounds during
storage had a far pro-
nounced effect on sur-
vival of HHP-treated
cells than ethanol.
Hop extract resulted
in sublethal injury of
virtually all cells with-
in 3 h of storage.
Viable cell counts
were reduced by 90%
within 3h and
dropped below the
detection level after
27 h of storage.
In commercial beer,
cell counts were
below the detection
level after 2 h.
200, 300, 400, 500 and 600 MPa, 15 C, various holding times MB with L. plantarum Similar results with Ulmer
et al.
that of Ulmer et al.
(2002)
(2000).
500 MPa, 15 C, 5 min MB with L. plantarum No inactivation but Ulmer
et al.
only sublethal injury
(2002)
at pH 6.0 and 7.0.
Higher inactivation at
pH b 5.0.
300 MPa, 15 C, 5 min MB (50 mg L1 hop extract) with L. plantarum Complete inactivation Ulmer
+ et al.
within 24 h at a stor-
MB (50 mg L1 hop extract and 5% ethanol) with L. plantarum (2002)
age temperature of
10 C.
100 MPa and 200 MPa, 15 C, 5 and 10 min MB (50 mg L1 hop extract) with L. plantarum L. plantarum was more pressure Ulmer
+ tolerant than S. uvarum. et al.
MB (50 mg L1 hop extract) with Saccharomyces uvarum (2002)
300 MPa, 20 C, various holding times up to 120 min MB with L. plantarum More effective inacti- Fischer
vation at lower pH et al.
(2006)
values.
Sublethal injury at
short holding times.

acid and 5.4 log10 acetic acid bacteria) were also treated at 400 MPa for
20 s, 1 and 2 min at RT. All lactic acid bacteria were destroyed even in
20 s whereas 2 min at the same pressure level was needed for complete
inactivation of acetic acid bacteria in red wines.
Different wines (white dry wine, liqueur-like white wine, ros
wine, a wine with the disease of grease and two other wines contam-
inated with Acetobacter aceti) were also treated at 300 MPa for 20 s,
1 min and 6 min and 400 MPa 20 s and 1 min at RT by Tonello et al.
(1996a). Treatment of 300 MPa for 20 s was sufcient to eliminate
2.5 and 3.5 log10 of S. cerevisiae present in white dry wine and
liqueur-like white wine, respectively. Same treatment was also suc-
cessful to eliminate 3.7 log10 of S. ludwigii in ros wine and 3.4 log10
of Pediococcus damnosus in wine with the disease of grease. For the
wines contaminated with A. aceti (one had 3.7 log10 and the other
10 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112
had 4.2 log10) treatments of 300 MPa for 6 min and 400 MPa for 20 s
and 1 min could be used for complete inactivation.
Tonello, Largeteau, Demazeau, and Lonvaud-Funel (1996b) ap-
plied HHP to inactivate different commercial strains of S. cerevisiae
in wines. HHP treatment at 300 MPa, 20 C for 20 s, 1 min and 6 min
showed that there was a variation in resistance to HHP among the
strains of S. cerevisiae in wines: one strain was completely eliminated
(7 log10) whereas reduction was only about 1.7 log10 for the most re-
sistant strain after 20 s at 300 MPa. Treatment at 300 MPa for 6 min
resulted in complete inactivation for all strains of S. cerevisiae. Most
resistant strain was used to show the effect of alcohol [8.6, 10.8, 13
and 15% (v/v)] and sugar (50% of glucose and 50% of fructose) during
HHP treatment of wines. Treatments were again applied at 300 MPa,
20 C for 20 s, 1 min and 6 min. Treatment of 6 min resulted in com-
plete elimination (7 log10) of the most resistant yeast in wine at all al-
cohol levels. Treatments of 20 s and 1 min revealed that yeast in
wines with alcohol levels of 8.6, 10.8 and 13% had similar inactivation
values whereas much higher inactivation was observed for wine con-
taining 15% of alcohol. There was no difference in inactivation of yeast
when sugar at the concentrations of 25 g L 1 to 98 g L 1 was added
to wine containing 13.3% of alcohol.
White, red and ros wines each inoculated with either yeast
(S. cerevisiae or S. ludwigii), lactic acid bacteria (Pediococcus) and acetic
acid bacteria (A. aceti) were also treated at 300 MPa for 20 s, 1 min and
6 min and 400 MPa for 1 min by Tonello et al. (1996b). 5 log10 reduction
of yeasts and Pediococcus was possible at 300 MPa for 20 s. A. aceti was
pressure resistant than lactic acid bacteria and yeasts; 300 MPa for
1 min treatment resulted in less than 2 log10 reduction. Treatment of
400 MPa for 1 min resulted in 5 log10 reduction of A. aceti.
Tonello, Largeteau, Demazeau, and Lonvaud-Funel (1998) studied the
HHP (180300 MPa for various times at RT) inactivation of S. cerevisiae in
wines. Mixture (50/50) of glucose and fructose and ethanol were used to
adjust sugar and alcohol content of wines. HHP treatment at 300 MPa for
20 s and 1 min showed no signicant inactivation levels between 8.6, 10.8
and 13% (v/v) alcohol content wines. Nevertheless, there was a signicant
difference between 8.6, 10.8 and 15% alcohol content wines. The alcohol
content increased the yeast inactivation but only when it was greater
than 13%. About 3 and 5 log10 reductions of yeast in 15% alcohol content
wine were observed at 300 MPa for 20 s and 1 min, respectively. Com-
plete elimination (about 7 log10) of yeasts in all wines was observed
after 6 min pressurization at 300 MPa. There was no difference in inacti-
vation of yeast when wine was fortied with sugar between 8 g L1
and 98 g L1. Wines made with S. cerevisiae after 9 days (D+9) or
18 days (D+18) of fermentation which were respectively in the middle
of stationary phase and the beginning of decline phase were also pressur-
ized. D+9 wine contained 8.9% alcohol and 1.5 g L1 sugar and D+18
wine had an alcoholic content of 9.1% and a sugar content of 0.9 g L1.
The results showed that barosensitivity of D+18 yeast was higher than
that of D+9 yeast.
Puig, Vilavella, Daoudi, Guamis, and Mnguez (2003) investigated
the microbiological and biochemical stabilization of wines by use of
HHP. A white wine (with 4050 mg L 1 of total SO2) and a red wine
(with 8090 mg L 1 of total SO2) were used. Two yeasts: S. cerevisiae
and Brettanomyces bruxellensis (10 7 CFUL1 of each wine), two lactic
acid bacteria: L. plantarum and Oenococcus onei (10 9 CFUL1 of each
wine), two acetic acid bacteria: A. aceti and A. pasteurianus
(109 CFUL 1 of each wine) were inoculated into wines. HHP treat-
ments were done at 400 or 500 MPa for 5 or 15 min with 4 C or 20 C
of temperature. HHP treatments resulted in almost complete inactiva-
tion (6 log10 reduction for yeasts and 8 log10 reduction for bacteria) of
the microorganisms for all conditions.
Sugar addition (15, 25 and 35 g L 1) to white wine had no baropro-
tective effect on yeasts and bacteria. HHP treatments did not affect poly-
phenoloxidase activity, alcohol level, total and volatile acidity, free and
total SO2, protein stability, malic acid, lactic acid, reducing sugars and
pH compared to untreated wines. Sensory evaluation was done with
12 series of tasting using 8 tasters. Tasters detected plastic taste in sam-
ples of three series which was due to the plastic container used in HHP
treatment. Other than this observation no organoleptic differences was
found between untreated and HHP-treated wines.
Puig et al. (2003) also compared HHP with ltration in terms of mi-
crobiological and biochemical stabilization. 10 different batches of wine
(red and white) with their natural microbial ora were used. Sugar
was added to one batch of white wine with concentrations of 15, 25
and 35 g L1. Three stabilization techniques were applied to wines:
(A) Kieselguhr ltration; (B) Kieselguhr ltration +ltration through a
module lter; (C) Kieselguhr ltration +HHP (500 MPa, 4 C for
7 min). Among the three treatments, the one with HHP produced the
highest reduction of yeasts. Refermentation, due to the activity of yeast
present in wine, was observed in samples treated by A and B. No signif-
icant differences in colorimetric parameters and in phenolic components
were observed in 10 batches of wine treated with three techniques. Poly-
phenoloxidase activity was undetectable in all cases. Sensory analyses of
red wine indicated that treatments B and C had positive impact in major-
ity of the cases compared to treatment A, whereas for the white wine di-
versity according to the type of wine (Macabeu, Chardonnay, Xarel-lo)
was observed.
Mok et al. (2006) studied the pasteurization of low-alcohol red
wine (9% ethanol, pH 3.27, acidity 0.068%, total sugar 0.851%) using
HHP (100350 MPa for 0 to 30 min at 25 C). There were about 5.6
log10 microorganisms (total aerobes) and about 5.46 log10 of both
yeasts and lactic acid bacteria in wine before HHP treatment.
The original microbial count (5.6 log10) decreased to 3.38, 3.07,
2.85, and 2.3 log10 after HHP treatment at 250 MPa for 5, 10, 20 and
30 min, respectively. Initial microbial count reduced to 2.44 and
1.82 log10 after HHP treatment at 300 MPa for 5 and 10 min, respec-
tively. The aerobic bacteria decreased below the detection limit after
20 min pressurization at 300 MPa and 10 min pressurization at
350 MPa. Initial yeast count (5.46 log10) decreased to 2.46 and 1.15
log10 after HHP treatment at 300 MPa for 10 and 20 min, respectively.
300 MPa for 30 min treatment killed all the yeasts in wine whereas all
lactic acid bacteria (initial count of 5.46 log10) inactivated at 300 MPa
for 20 min or 350 MPa for 5 min.
Sensory evaluation done using 10 trained panels revealed that
there were no differences in the aroma, taste, mouth-feel and overall
sensory quality between the HHP-treated (350 MPa for 10 min) and
untreated samples.
Corrales, Butz, and Tauscher (2008) applied HHP to wine from the
Dornfelder grape variety. The stability of the predominant anthocya-
nins such as malvidin-3-O-glucoside (Mv3gl) in wine was monitored.
A decrease in the concentration of Mv3gl in pressurized (600 MPa at
70 C for 1 h) samples was detected whereas, in the heat-treated
(0.1 MPa, 70 C for 1 h) samples, no loss of Mv3gl was observed com-
pared to untreated sample (0.1 MPa, 20 C for 1 h). When wine was
subjected to 600 MPa, 70 C for 10 min, no signicant differences in
anthocyanin composition or antioxidant activity were found.
Table 2 summarizes the studies done about wine.
4. Concluding remarks and future perspectives
Although very few studies on beer and wine with HHP are avail-
able today, the potential of HHP technology is huge in both beer
and wine industries: Studies have shown that HHP treatment not
only inactivates the undesirable microorganisms but also improves
the organoleptic properties of beer and wine. The pressure levels
used to treat beer and wine were similar to the commercial applica-
tions used in fruit juice industry i.e., 400600 MPa.
It should be noted that installation of an HHP equipment to a
brewery or a winery would denitely bring an extra cost; however,
the HHP-treated product would have a fresh-like taste and would
most probably attract the consumers' attention. Although there is
no sensory evaluation of HHP-treated beer in literature, in case of
 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112 11

Table 2
Applications of high hydrostatic pressure (HHP) on wine.

Pressure, temperature, time Sample treated Achievements Reference

200400 MPa, various holding times
300600 MPa, 20 C
300 and 400 MPa, room
temperature, various holding times
300 MPa& 400, 20 C, various holding Wines contaminated with S. cerevisiae or S. ludwigii,
times
180300 MPa, room temperature,
various holding times
400 or 500 MPa, 4 C or 20 C, 5 or
15 min
100350 MPa, 25 C, 030 min
600 MPa, 70 C, 1 h
+ et al.
600 MPa, 70 C, 10 min
Sauternes wine Complete elimination of yeasts. Lonvaud-
Funel et al.
No change in viscosity.
(1994)
Color values of untreated wine was higher than
that of HHP treated wine samples.
Oxidation took place in HHP treated samples during
storage.
Moscato wines obtained from Barbera grape must+ HHP had a strong antimicrobial effect. Delni
Asti Spumante wine et al.
No variation of the color parameters compared
(1995)
with the untreated wine.
White, red, ros wines+ Complete elimination of yeasts. Tonello
Liqueur-like white wine+ et al.
Ethanol addition accelerated inactivation of yeasts
Wine with the disease of grease+ (1996a)
Wines contaminated with Acetobacter aceti whereas sugar addition had no effect.
Complete elimination of lactic and acetic acid
bacteria.
Complete elimination of A. Aceti.
Variation in resistance to HHP among the strains of Tonello
Pediococcus and A. aceti et al.
S. cerevisiae in wines & complete elimination of
(1996b)
S. cerevisiae.
Ethanol addition accelerated inactivation of yeasts
whereas sugar addition had no effect.
A. aceti was pressure resistant than lactic acid
bacteria and yeasts.
Wines with S. cerevisiae Similar results with that of Tonello et al. (1996a; Tonello
et al.
1996b)
(1998)
White and red wines Complete inactivation of yeasts, lactic and acetic Puig et al.
(2003)
acid bacteria.
No baroprotective effect of sugar addition on
yeasts and bacteria.
No effect on polyphenoloxidase activity, alcohol
level, total and volatile acidity, free and total SO2,
protein stability, malic acid, lactic acid, reducing
sugars and pH.
No organoleptic differences between untreated
and HHP-treated wines.
Kieselguhr ltration + HHP produced the highest
reduction of yeasts compared to Kieselguhr ltra-
tion only and Kieselguhr ltration + ltration
through a module lter.
Low-alcohol red wine Complete inactivation of yeasts, lactic and acetic Mok et al.
(2006)
acid bacteria.
No differences in the aroma, taste, mouth-feel and
overall sensory quality between the HHP-treated
and untreated samples.
Wine from the Dornfelder grape variety Decrease in the concentration of malvidin-3-O- Corrales
glucoside.
(2008)
No differences in anthocyanin composition or an-
tioxidant activity.

beer the negative effect of heat on the aroma and avor beer could be
eliminated by use of HHP. HHP-treated beer and untreated beer
might have the similar taste. On the other hand, although SO2 can
have negative effects on human health it acts as both an antimicrobial
agent and an antioxidant in wine. Nevertheless, use of HHP could help
the wine industry to reduce the levels of SO2 or HHP could be used in
combination with other antimicrobial agents such as nisin.
Future studies may focus more on sensory evaluations since HHP
proved itself to destroy spoilage organisms in beer and wine. The
cost analyses of the treatment with respect to the capacity of the pro-
duction and also the product itself should be performed. Moreover,
surveys should be done to investigate the consumer awareness and
willingness to pay HHP-treated beer or wine.
Acknowledgments
Author thanks Prof. C. Prez-Lamela and Prof. R. Vogel for supply-
ing their studies about beer and Mr. Erkan Frat Yaar for his help to
compile the some of the references used in this contribution. This
study is dedicated to the employees of Alcoholic Beverages Market
Department in TAPDK.
12 S. Buzrul / Innovative Food Science and Emerging Technologies 13 (2012) 112

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