Impact of maceration time on colour-related phenolics, sensory characteristics and volatile

composition of mulberry wine

Introduction
Mulberry, also known as Morus nigra L., is a perennial woody plant that is widely grown in
India, China, Japan, North Africa and South- ern Europe (1–3). Mulberry fruits contain
numerous bioactive com- pounds, including polysaccharides, anthocyanins, flavonoids and
phenolic acids (1,4,5). These bioactive ingredients play important roles in providing mulberry
fruits with numerous beneficial fea- tures, such as anti-inflammatory, anti-atherogenic and
anti- oxidative properties (5–8). Mulberry fruits possess a thin skin and juicy pulp,making
themsusceptible to spoilage during the storage and transportation process (9). In order to
extend their shelf-life, mulberry fruits are normally processed into mulberry juice, jam, syrup
and wine (10). Among these products, mulberry wine has gained much attention in the wine
industry of China owing to its appearance and sensory attributes (9).
Phenolic compounds are extracted from the skin and pulp of mulberry to wine during the
fermentation process, and they con- tribute to the sensory properties of mulberry wines,
together with appearance and mouthfeel. For example, anthocyanins provide mulberry wine
with favourable colour (9,11,12), whereas the bitter- ness and astringency of mulberry wine
results mainly from non- anthocyanin phenolic compounds (13). The evolution of phenolic
compounds inmulberrywineduring fermentationandstoragepro- cess also alter the appearance
of mulberry wine (12,14,15). For ex- ample, non-anthocyanin phenolic compounds during
fermentation process can interact with anthocyanins through hy- drophobic interactions and
hydrogen bonds, which stabilizes the structure of anthocyanins inmulberry wine (16–18).
Polymerization betweenanthocyaninsandother phenoliccompounds inwinedur- ing storage
results in theprecipitationofanthocyanins, which shifts the wine colour from red towards brown
(19,20).
analysis of phenolic compounds in pomace extracts from five grape cultivars: evaluation of
their antioxidant, antiradical and antifungal activities in orange and apple juices.
Volatile compounds, on the other hand, contribute to the overall aroma ofmulberry wine
(21,22). It has been reported that the over- all aroma in wine consists of fruit-derived volatiles,
fermented vol- atiles and the volatiles produced during wine aging process (23,24). The fruit-
derived volatiles are present in mulberry fruit and they are extracted during the maceration
period (24,25).The mulberry wine fermentation process produces numerous volatile
compounds, such as alcohols, aldehydes and ketones, and these volatile compounds can
improve the fermented aroma (24,26). The evolution of volatile compounds during the wine
aging pro- cess can introduce an aging aroma which can further enhance the wine maturation
(27). The fruit-derived volatiles in wine essen- tially determine the variety flavour.
During wine fermentation, maceration is considered an important process by which phenolic
and volatile components in fruits can be extracted into wine (28,29). In has been confirmed
that mac- eration time plays a primary role in affecting the appearance and sensory attributes
(9,30,31). For example, a shorter maceration treatment reduces the extraction efficiency of
these components from fruit skins, which weakens the sensory properties of wine (32–34).
However, macerating fruit skins for a longer period during wine fermentation has been
reported to induce undesired com- pounds into wine, lowering the wine sensory quality (35).
The effect of maceration treatment on grape wine quality has been widely studied. However,
mulberry is different fromgrape in fruit and seed structure, and accordingly the chemical
nature of mulberry is not similar to grape. More importantly, no study has been carried out on
the maceration effect on phenolic and volatile compounds in mulberry wine to our best
knowledge. Therefore, we evaluated three different maceration periods (2, 6, and 8 days)
during the mulberry wine fermentation process. Phenolic and volatile profiles, colour attributes
and the overall aroma features ofmulberry wines from these maceration treatments were
analysed and compared. We aimed to investigate if different maceration treatments signifi-
cantly affected the appearance and sensory attributes of mulberry wine, which could provide
insight for the improvement of mul- berry wine production.
Cold maceration application in red wine production and its effects on phenolic compounds: A
review
Introduction
The so-called maceration step i.e. the period during the red winemaking process in which the
solid parts of the berries are in contact with the must is of crucial importance. A number of key
compounds are transferred from the skins and seeds into the must while the contact is
maintained. Among them, relevant phenolic compounds are extracted. The reason why
phenolic compounds are important is because they define some of the most significant
organoleptic attributes in red wines (Aleixandre-Tudo, Buica, Nieuwoudt, Aleixandre, & du Toit,
2017; Casassa & Harbertson, 2014). Visual red wine properties are mostly due to anthocyanins,
but most importantly by their interactions, reactions and combinations among themselves or
with other wine substances (He et al., 2012). Furthermore, proanthocyanins or tannins are
relevant compounds responsible for some of the most appreciated mouthfeel attributes, i.e.
bitterness and astringency. In this case the molecular size (degree of polymerization), subunit
composition and combinations with other wine components influence wine’s mouthfeel
perception (Smith, Mcrae, & Bindon, 2015)
Due to the importance of the maceration step with regards to final product quality, a number
of winemaking practices are commonly used to optimize the extraction/diffusion of phenolic
substances into the wine. The application of a maceration period before the start of the
fermentation, the so-known cold maceration (CM) or cold soak, is one of the most applied and
investigated practices. The reasoning behind the technique states that during the pre-
fermentative period phenolic compounds are transferred from the solid parts of the berry into
the must under low temperatures (to prevent the start of the fermentation) and in an alcohol-
free environment (aqueous media). In theory, as a result of the extended contact period, a
higher amount of phenolic compounds will finally be present in the wine. Despite the
aforementioned reasoning the results found in the literature are variable when applying this
pre-fermentation technique. Due to its economical, logistic and quality implications it is
therefore important to understand the factors that influence the effectiveness of the technique
before deciding on a CM application.
Release of phenolic compounds from the solid parts of the berries during skin contact
An increase in the contact time between must and solid parts of the berries is the main
purpose of the CM practice. In order for this technique to be effective the contact period needs
to happen under low temperatures to avoid yeast growth and fermentation development. The
extraction/diffusion of phenolic compounds thus occurs in the absence of ethanol (Sacchi,
Bisson, & Adams, 2005). Phenolic compounds are water soluble molecules, however, phenolic
substances are mostly located in the vacuoles of the grape cells and in order for them to be
released, the cell wall needs to be broken, degraded or at least disorganized (increased
permeability) (Gil-Muñoz et al., 2009).
Skin phenolic compounds include flavan-3-ols and proanthocyanidins (the so-called skin
tannins), anthocyanins and flavonols. Anthocyanins are found in cell vacuolar inclusions located
in the first layers of the epidermal tissue. A two-step extraction/diffusion process is thus
required. The first biological barrier corresponds to the membrane bound vacuole and the cell
itself while the second barrier has to do with the cell wall (Casassa & Harbertson, 2014). The
crushing operation ensures the breakdown of the vacuole and cell membranes, 7 while
extraction takes place after cell wall degradation, enhanced , by enzymatic activity on pectin
and polysaccharides related cell wall components (Renard, Watrelot, & Le Bourvellec, 2017).
Moreover, the diffusion is favoured due to the water soluble nature of these compounds. In
addition, the early extraction and subsequent decrease of the anthocyanins during winemaking
indicates that the final levels found in wines are predominantly related to its instability rather
than to solubility conditions (Casassa & Harbertson, 2014). It has also been reported that in
order to efficiently release internal components, the cell membrane needs to be broken and
not just collapsed as in the latter case the membrane structure 7 might retain phenolic
components (Sacchi et al., 2005; Watrelot, Le Bourvellec, Imberty, & Renard, 2013). Flavonols
are also vacuolar substances thus co-located with the anthocyanins; however, the extraction
pattern of these compounds, although similar to that of anthocyanins, is slower, mainly due to
polarity conditions (Casassa & Harbertson, 2014).
The extraction of flavan-3-ols, located in the inner-thick cells of the skins hypodermis, occurs
during the early stages of skin contact. The diffusion patterns of proanthocyanidins (PAs)
dimers and trimers mimic the extraction behaviour of their monomeric counterparts.
Moreover, the extraction rate of oligomeric and polymeric PAs seems to progress slower as skin
contact time increases (Casassa & Harbertson, 2014). It has also been reported that chemical
properties (hydrophobicity) and composition (number of hydroxyl groups) might influence the
extraction kinetics (Koyama, Goto-Yamamoto, & Hashizume, 2007). On the
The investigation of phenolic compounds and anthocyanins of wines made of the grape
variety karmrahyut
Introduction
Phenolic compounds can be commonly classified into five groups: anthocyanins, flavan-3-ols,
flavonols, stilbene derivatives and phenolic acids which are considered to possess the ability to
scavenge excess radicals and maintain the balance of reactive oxygen species (ROS) in the
human body [4,5]. According to many authors, antioxidant activity of grape berries and wines
results mainly from their phenolics, whereas the phenolic content and composition depend on
the grape variety, vineyard location, cultivation system, climate, soil types, vine cultiva- tion
practices, harvesting time, production process and ageing [1,3]. Anthocyanins, also known as
anthocyans, are water soluble flavonoid pigments that, depending on pH, and in some cases
complexing agents, can contribute diverse colors such as red, purple and blue. They are widely
spread throughout the plant kingdom [6,7]
Color is one of the most important attributes of red wines, and the principal sources of red
color in wines come from the anthocyanins or their further derivatives that are extracted or
formed during the vini- fication process [2,8].
Normally, anthocyanins are mainly located in the grape skins, with a few exceptions, in the so-
called “teinturier” grapes, which have an- thocyanins in both of the skin and the pulp [9–11].
Hybrid red grape varieties have significantly higher concentration of phenolic com- pounds
than the industrial grape varieties. They have also higher an- tioxidant activity [12]. During
fermentation and, especially in the firs one or two years of maturation, the anthocyanins in
wines undergo a wide variety of reactions and associations and various anthocyanin- derived
new pigments are formed, which are extremely crucial for the color stability. Consequently,
although the concentration of anthocya- nins in red wines declines constantly, red wines can
still maintain an essentially red color. The reactions and associations involve complex
mechanisms, including relatively short term ones, such as self-associa- tion and
copigmentation, and the relatively long-term ones, such as the formation of polymeric
anthocyanins, such as pyranoanthocyanins and their further polymerized products [13–15].
Effect of maceration duration on physicochemical characteristics, organic acid, phenolic
compounds and antioxidant activity of red wine from Vitis vinifera L. Karaoglan
Introduction
Grape (Vitis vinifera L.) contains a number of nutritional and functional compounds and it is
rich in phenolic com- pounds. A number of phenolic compounds are present in seed and skin of
grape. It was reported that the total extractable phenolics mainly distributed in skin (28–35 %)
and seeds (60–70 %) in comparison to the pulp (10 %) in fresh grapes (Sagdic et al. 2011) and
the phenolics in skin are transferred into the final product by the maceration process in red
winemaking. In particular, red wine is rich in phenolic compounds, including flavonoids
(anthocyanins, flavan-3-ols, proanthocyanidins or condensed tannins and flavanols) and non-
flavanoids (hydroxybenzoic and hydroxycinnamic acids and their derivatives, stilbenes and
phenol alcohols) based on grape variety, growing tech- niques (viticulture) and winemaking
conditions (Budic- Leto et al. 2008; Ginjom et al. 2011). In this context, the qualitative and
quantitative properties of phenolics in grape are affected by ripening stage of grape, climate,
soil, growth area of grape and winemaking conditions including maceration duration,
temperature, pressing intensity, inoculation level and type of yeast, the amount of sulfur
dioxide, and other practices (Ivanova et al. 2011). The phenolic compounds (e.g., anthocyanins,
stilbens, flavan-3- ols) are important for a high quality of red wine and these contribute to
antioxidant activity which is associated with some health benefits (Lucena et al. 2010). It was
reported that the moderate consumption of red wine has some bio- logical activities such as
cardioprotective effects, preven- tion of atherosclerosis, coronary heart disease, anti-
inflammatory responses, prevention of low density lipoprotein oxidation, antihypertensive and
carsinostatic properties, etc. (Lucena et al. 2010). Phenolic compounds also affect the sensory
characteristics of wine, in particular color as a result of their interactions with colorless phe-
nolics, polysaccharides, metals and anthocyanins. Colored (anthocyanins) and colorless
phenolic compounds are extracted from grape skins into wine by means of macer- ation at a
controlled temperature and time (Kelebek et al. 2006; Hernanz et al. 2007). Maceration
process may cause considerable variations in wine quality depending on grape cultivar
employed. It was reported that the prolongation of the maceration time greatly increases the
levels of phenolic compounds and improves the color stability of wine (Kelebek et al. 2009).
Karaoglan is an autochthonous cultivar of Vitis Vinifera L. grown in Arapgir, county of Malatya
(Turkey). This aromatic variety has a round shaped grapes with a thick skin. It is traditionally
used for table grape consumption; however, it has been recently used for red winemaking due
to its dark color and high aroma potential. Karaoglan wines have a typical aroma characterized
by fruit (raspberry, cherry, and strawberry) flavours. Although volatile and sensory
characterization of Karaoglan red wines have been made in some extent (Yilmaztekin et al.
2015); however, chemical composition, phenolics and antioxidant charac- teristics has not yet
been studied. The objective of the present study was to identify and quantify some physical
properties, chemical composition and individual phenolic compounds of Karaoglan red wine
and to investigate the effects of varying maceration duration on phenolic com- pounds and
antioxidant activities of wine.
Modulating analytical characteristics of thermovinified Carignan musts and the volatile
composition of the resulting wines through the heating temperature
1. Introduction
Pre-fermentation heat treatment of grapes or thermovinification is a winemaking technology
first industrially developed in the seventies (Marteau & Olivieri, 1970). It consists of heating
grapes between 70 and 75 °C for a length of time varying from 30 min to 24 h. The term
“thermovinification” is sometimes used to describe the process in which heating is limited to a
brief period (< 1h). After pressing at a high temperature and clarification, fermentation is
usually undertaken in liquid phase at a lower temperature than usual red ferments, typically
between 18 °C and 25 °C. This technique is becoming increasingly popular for the production of
colored, fruit driven red wines with soft tannins. The volume of wine elaborated in France in
2017 through thermovinification was estimated at 750 million liters (J.L. Favarel, personal
communication, July 18, 2017).
In comparison with control macerated wines, thermovinification wines fermented at a lower
temperature usually have higher levels of ethanol (Geffroy et al., 2015). Some changes in the
acid–base balance of the wines by higher tartaric acid and potassium extractions from the
pericarp tissue of the berries were also highlighted. The typical sensory profile of
thermovinified wines is due to a large extraction under the effects of heat of hydrophilic
anthocyanin pigments and grape poly- saccharides responsible for roundness in wine (Doco,
Williams, & Cheynier, 2007; Girard, Kopp, Reynolds, & Cliff, 1997). In aqueous phase, the
extraction of tannins is more moderate and the technique usually leads to wines with a lower
overall phenolic content than those made using traditional maceration techniques (Auw,
Blanco, O'Keefe, & Sims, 1996).
From an olfactive point of view, thermovinification is known to produce wines with a
standardized sensory profile often described as “banana yogurt” by winemakers. Past research
into the volatile composition of thermovinified wines showed that maceration heat treat- ment
allowed the elimination through volatilization of a large amount of 3-isobutyl-2-
methoxypyrazine (Roujou de Boubée, 2000) and was not favorable to the production of wines
with high concentrations of hydrophobic rotundone (Geffroy, Siebert, Silvano, & Herderich,
2016). Another work reported the likely thermal degradation of several grape- derived aroma
compounds or their precursors (i.e. some varietal thiols, monoterpenols, norisoprenoids,
phenols) when grapes were heated during 3 h at 70 °C (Geffroy et al., 2015). The fermentation
conditions of thermovinified wines particularly enhanced esters, acetates and fatty acid
formation (Cottereau & Desseigne, 2007; Fischer, Strasser, & Gutzler, 2000; Girard et al., 1997).
Consistent with previous observa- tions made in the seventies (Poux, 1974), Geffroy et al.
(2015) recently showed that heating at 70 °C for 2 h followed by pressing at a high temperature
induced a substantial increase in the concentration of amino acids in the must (from+101% to
200%). The fermentation in liquid phase and at low temperature of high Yeast Assimilable
Nitrogen (YAN) clarified musts enhanced the production of fermentative aroma compounds by
the yeast (Moreno, Medina, & Garcia, 1988).
The modulation of the sensory profile of thermovinified wines towards a fruiter varietal
character is an issue frequently raised by winemakers. Grape derived aroma compounds
imparting this character to the wine include monoterpenes, norisoprenoids, aliphatics, phenyl-
propanoids, methoxypyrazines, and volatile sulfur compounds (Robinson et al., 2014).
In an attempt to produce thermovinified wines with a fruiter varietal character, two levels of
must clarification (150 and 800 nephelo- metric turbidity units or NTU) and fermentation
temperature (18 °C and 25 °C) were previously investigated (Geffroy et al. (2014)). The results
were inconclusive as both factors had an overall weak impact on the aroma composition and
sensory profile of the wines. However, wines fermented at 25 °C were judged slightly less
amylic and more complex. Minor differences were observed between wines made from distinct
clarification levels, and these were limited to mouthfeel and taste perception.
In relation with amino acid extraction, thermal degradation and volatilization of aroma
compounds, the heating temperatures could be adjusted to modulate the volatile composition
of thermovinified wines. Thermovinification was originally used on botrytized grapes to destroy
laccase whose activity increases with temperature up to its denatura- tion point of 60 °C. This is
the main reason why the heating of rotten grapes must be done very quickly at a temperature
above 70 °C (Ribéreau-Gayon, Dubourdieu, Donèche, & Lonvaud, 2005). Nowadays
thermovinification is mainly employed on grapes of perfect sanitary status without Botrytis
cinerea and new ranges of temperature, espe- cially below 60 °C, deserve to be investigated. As
a decrease in the heating temperature is likely to impact the level of phenolic compounds in
wine, the heating time would need to be adapted.
The purpose of the present work is to study the impact of the temperature and the heating
time on the analytical and volatile com- position of thermovinified musts and wines. In 2014
and 2015, two temperature levels (50 °C and 75 °C) and heating times (30 min and 3 h) were
investigated in duplicate at the laboratory scale on Carignan grapes sourced in Spain.
Pre-fermentative cold maceration, saignée, and various thermal treatments as options for
modulating volatile aroma and phenol profiles of red wine
Introduction
Volatile aromas and phenols are two of the most important groups of chemical compounds
that determine the sensory quality of red wine (Álvarez, Aleixandre, García, & Lizama, 2006;
Sáenz-Navajas, Avizcuri, Ballester, Fernández-Zurbano, Ferreira, Peyron, & Valentin, 2015).
Aroma compounds originate from grapes, and are formed in fermentation and during wine
maturation. They pertain to different chemical classes, such as monoterpenes, norisoprenoids,
higher alcohols, fatty acids, esters, aldehydes, ketones, etc. (Callejón, Margulies, Hirson, &
Ebeler, 2012). Phenolic compounds are mainly extracted from grapes during maceration (skin
and seed contact), and are responsible for several important sensory properties of red wine,
such as taste, mouthfeel and color (González-Neves, Favre, Gil, Ferrer, & Charamelo, 2015;
Hufnagel & Hofmann, 2008). Among them, phenolic acids, flavonols and tannins participate in
astringency and bitterness (Garrido & Borges, 2013; Hufnagel & Hofmann, 2008), while red
wine color is mostly determined by anthocyanins (Garrido & Borges, 2013; González-Neves et
al., 2015).
Maceration conditions, such as duration and temperature, are the most important factors that
modify the diffusion of varietal aromas and phenols from solid grape parts and their
solubilization into must and wine (Sacchi, Bisson, & Adams, 2005). Maceration parameters also
affect the formation and levels of fermentation aromas, although to a lesser extent (Callejón et
al., 2012). Various maceration procedures in the production of red wines have been studied to
establish the selective effects of different parameters on the extraction of important grape
components. Several treatments, such as cold-soak pre-fermentative maceration, carbonic
maceration, and delestage, exhibited contradictory or mixed results, mainly depending on the
variety (Sacchi et al., 2005). Other techniques, developed earlier but revisited today in a
completely different context, such as extended maceration (Baiano, Terracone, Gambacorta, &
La Notte, 2009), saignée (Harbertson, Mireles, Harwood, Weller, & Ross, 2009), and maceration
with heating (Baiano et al., 2009), were not studied extensively, and their effects are practically
unknown. For example, it is estimated that in France currently 500 millions of liters of red wine
are produced by short pre-fermentative heat treatment and liquid juice fermentation (Geffroy,
Lopez, Serrano, Dufourcq, Gracia-Moreno, Cacho, & Ferreira, 2015), however, very few studies
have investigated the effects of this technique.
In general, a small number of studies directly compared the effects of a larger number of
maceration procedures on the same grape material (Baiano, et al., 2009; Harbertson et al.,
2009; Mihnea, González-San José, Ortega-Heras, & Pérez-Magariño, 2015), while the majority
of further studies were often limited in that they either confronted a relatively small number or
mostly standard techniques. Their relative effectiveness could often be assessed only on the
basis of the comparison of results from different sources, which could lead to misleading
conclusions. Finally, a rather limited number of investigations were comprehensive in a way
they studied the effect of maceration on both aromas and phenols (Álvarez, Aleixandre, García,
& Lizama, 2006; Fischer, Strasser, & Gutzler, 2000; González- Neves et al., 2015), which are
inseparably related, and complementary in the context of wine sensory quality.
For all the mentioned reasons, the aim of this work was to evaluate the effect of certain
common, and particular emerging maceration techniques as alternatives to standard
procedures, on the volatile aroma and phenol composition of red wine: cold pre-fermentative
fermentation, saignée, pre-fermentative heating followed by both traditional fermentative
maceration and fermentation of juice, and post-fermentative heating. Total duration of all
treatments except juice fermentation was extended to 20 days, to annul the effect of
maceration duration. Equating the duration of treatments was expected to more precisely
determine the effects of low temperature in pre-fermentation cold maceration treatment in
relation to previous research, where in most cases, it involved an additional skin contact time in
relation to the control (Cai, Zhu, Wang, Lu, Lan, Reeves, & Duan, 2014; González-Neves et al.,
2015; Mihnea et al., 2015)