Food Chemistry 126 (2011) 574–582

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Influence of pollen addition on mead elaboration: Physicochemical

and sensory characteristics
A. Roldán a,⇑, G.C.J. van Muiswinkel b, C. Lasanta a, V. Palacios a, I. Caro a
a
Department of Chemical Engineering and Food Technology, Cádiz University, Aptdo 40, Polígono Río San Pedro, 11510 Puerto Real, Cádiz, Spain
b
Department of Agrotechnology and Food Sciences, Wageningen University and Research Center, Bomenweg 2, 6703 HD Wageningen, The Netherlands

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

Article history: Mead (honey-wine) results from the alcoholic fermentation of diluted honey using a wine yeast strain.
Received 3 May 2010 However, mead elaboration can be hampered by several problems, including delayed or arrested fermen-
Received in revised form 25 September 2010 tation, production of an unpleasant aroma, poor quality and inconsistency of the final product. These dif-
Accepted 10 November 2010
ficulties are due to honey’s low nutrient content, its natural antifungal components, and the inability of
the yeast strain to adapt to these unfavourable growth conditions. In this study, we evaluated the results
of adding pollen at concentrations ranging from 10 to 50 g/l as a fermentation activator to improve the
Keywords:
fermentation kinetic and the quality of meads. The effect of pollen addition on the honey must, fermen-
Mead
Honey
tation kinetics, physicochemical characteristics, aroma profiles and sensorial aspects of the obtained
Honey wine meads were evaluated. The results showed that pollen addition improved fermentation rates, alcohol
Fermentation yields, and the final characteristics of meads. An increase in the volatile contents of the meads and an
Pollen improved sensory profile was observed with pollen addition; however, this improvement was not corre-
Aroma profile lated with the concentration of pollen. The adequate dose of pollen was determined by the final charac-
teristics and sensory profile of the meads.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction
Honey, the sweet substance produced by honey bees, has been
used for centuries to prepare traditional, homemade drinks. Honey
can be fermented to produce different types of mead (honey wine),
sherry type wine, sparkling wine and fruit-honey wine, which may
have different flavours depending the floral source of the honey
and the additives and yeast used in fermentation (Gupta & Sharma,
2009).
Honey production has a significant economic importance in
several countries, and many scientific works about it have been
published (Baroni et al., 2009; Estevinho, Pereira, Moreira, Dias, &
Pereira, 2008;), and about the health benefits of honey (Bogdanov,
Jurendic, Sieber, & Gallmann, 2008; Sato & Miyata, 2000). However,
there are few scientific studies about mead and other honey prod-
ucts (Pereira, Dias, Andrad, Ramalhosa, & Estvinho, 2009; Sroka &
Tuszýnski, 2007).
All honeys share certain general characteristics, including a
moisture content below 20%, a sugar content of 70–80%, an ash
content ranging from 0.1% to 0.2%, and a pH between 3.8 and 4.7
(Nagai, Inoue, Kanamori, Suzuki, & Nagashima, 2006; Ouch-
emoukh, Louaileche, & Schweitzer, 2007; Silva, Videira, Monteiro,
⇑ Corresponding author.
E-mail address: ana.roldan@uca.es (A. Roldán).
Valentão, & Andrade, 2009). Proteins, free amino acids (principally
proline), organic acids, aromatics, and vitamins and minerals are
minor components (Hernández, Fraga, Jiménez, & Arias, 2005;
Pisani, Protano, & Riccobono, 2008). Honey also contains antimi-
crobial and antioxidant properties (Gomes, Leandro, Moreira,
Rodrigues, & Estevinho, 2010; Nagai et al., 2006).
Mead is an alcoholic beverage containing 8–18% (v/v) ethanol,
obtained by the alcoholic fermentation of diluted bee honey with
an appropriate amount of water. Fruits, juices and spices may also
be added (Gupta & Sharma, 2009). The time needed for fermenta-
tion and maturation ranges from several months to several years.
During that time, several problems can occur: fermentation may
be delayed or arrested; the yeast may referment; volatile acidity
can increase; bacteria may contaminate the mead, changing its
organoleptic quality; and the final product may lack uniformity
(Pereira et al., 2009; Sroka & Tuszýnski, 2007).
Experience with grape wine production indicates that these
problems are usually associated with the yeast strains’ inability
to adapt to unfavourable growth conditions, such as limitations
in nutrients, osmotic stress, ethanol toxicity and temperature
shock stresses (Attfield, 1997; Bauer & Pretorius, 2000; Bisson,
1999).
Yeast from the Saccharomyces cerevisiae strain, used in wine and
beer production, has also been used as a starter in mead produc-
tion. Recently, the capacity of S. cerevisiae strains, isolated from
Portuguese honeys, to produce mead has been evaluated by Pereira

0308-8146/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.11.045
 A. Roldán et al. / Food Chemistry 126 (2011) 574–582 575

et al. (2009). Under the stress conditions (ethanol, sulphur dioxide
and osmotic stresses) studied by these authors, the yeast strains
isolated from honey behaved similarly to the commercial wine
strains used in enology. That study also showed that the mead pro-
duction outcomes depend on the composition of the honey used
and the supplements added to it. Moreover, Sroka and Tuszýnski
(2007) showed that the acetic and succinic acids formed during di-
luted honey fermentation reduce the mead’s pH and lead to a in-
creased non-dissociated fatty acid content, which, in addition to
the presence of relatively large amounts of medium-chain fatty
acids, may cause the fermentation to slow down or stop.
The nutritional requirements of yeast for fermenting grape
must have been relatively well researched and described (Alexan-
dre & Charpentier, 1998; Bell & Henschke, 2008; Bisson & Butzke,
2000). These nutrients, particularly yeast-assimilable nitrogen
(YAN), are essential for the optimal growth and development of
the yeast (Barre, Blondin, Fuillat, Sablayrolles, & Salmon, 1998; Rib-
eréau-Gayón, Dubordieu, Doneche, & Louvand, 2000; Bell & Hens-
chke, 2008). However, the nitrogen level of the fermentation media
modifies the sensory character of mead (Vidrih & Hribar, 2007) be-
cause the amino acid composition affects the yeast’s metabolism
and thereby the production of the volatile aromatic compounds.
The amino acid compounds are precursors to fermentation (Bous-
eta, Scheirman, & Collin, 1996; Vilanova, Ugliano, Siebert, Pretori-
us, & Henschke, 2007).
The lower pH and the low mineral content of light honey versus
dark honey can decrease the growth of yeast (Pereira et al., 2009).
Fruit juices, salts and acids have been used as additives to stimu-
late the fermentation and improve the mead production process
(Gupta & Sharma, 2009). Formulations with different amounts of
ammonium phosphate, potassium sodium tartrate, magnesium
sulfate, calcium sulfate, citric acid, tartaric acid and vitamins (bio-
tin, pyridoxine, thiamin) have been used as supplements in many
studies (Gupta & Sharma, 2009; Pereira et al., 2009). In a previous
study by our research group (Roldán, van Muiswinkel, Lasanta, &
Caro, 2008), different grape juice fermentation activators were
studied to improve diluted honey fermentation, including thiamine
chlorhydrate, yeast extract, pollen and royal jelly (the last two of
which are derived from beehives). The results showed that pollen
improved the fermentation kinetics the most and was the best fer-
mentation activator. Pereira et al. (2009) showed that the greatest
difficulty in mead production came from attempting to ferment
honey with a low pollen content.
Pollen is collected from flowers and is the most important
source of proteins, lipids, minerals and vitamins for bee survival.
Recently, increasing evidence suggests its potential therapeutic
benefits, including antioxidant properties (Leja, Mareczek, Wyzgo-
lik, Klepacz-Baniak & Czekonska, 2007) and bioactive properties as
functional dietary food supplement (Kroyer & Hegedus, 2001). In
addition to sugars, pollen contains 7.4% moisture, proteins (at least
20%), 6% lipids and 2.2% ash, plus minerals, vitamins and carote-
noids (Almeida-Muradian, Pamplona, Coimbra, & Monika Barth,
2005; Human & Nicolson, 2006). Proline, aspartic acid, phenylala-
nine and glutamic acid are the primary amino acids in pollen (Gon-
zález, Gómez, Cordón, García-Villanova & Sánchez, 2006).
The aim of this work was to evaluate the influence of pollen
addition on mead elaboration. Different concentrations of pollen
were used as an activator to improve fermentation and the final
characteristics of meads. The fermentation kinetics, physicochem-
ical characteristics, aroma profiles, and sensory aspects of the final
products were determined.
2. Materials and methods
2.1. Mead elaboration

Others authors instructions were followed to produce mead

(Gupta & Sharma, 2009; Pereira et al., 2009; Vidrih & Hribar,
Table 1
Volatile compounds identified, odour descriptor, odour series and literature odour
2007). Commercial honey (Valencia, Spain) (75° Brix) of multifloral
threshold. origin (3 kg for each test) and always from the same marketing
firm was diluted with water until a diluted honey of approximately
Compound Odour descriptor Odour Series Threshold
12 °Be (20–22 °Brix) was obtained. The acidity was corrected to an
(OS)
approximate pH of 3.6 with tartaric acid, because according to
Isovaleric acid Acid, sour, cheese Oxidation (DO) 33 lg/la
Gupta and Sharma (2009), tartaric acid is less easily metabolised
Octanoic acid Oily, rancid, butter, Oxidation (DO) 500 lg/lb
cheese by undesirable lactic acid bacteria. Potassium metabisulphite was
Ethyl 3-hidroxy Racid Oxidation (DO) 67000 lg/lc added (up to 50 mg/l) to prevent lactic acid bacteria growth. The
butanoate diluted honey was divided into 5 l fermentation tanks, and com-
3-etoxy-1-propanol Fruity Fruity (F) 50000 lg/lc
mercially produced pollen (Valencia, Spain) was added in concen-
Isoamyl acetate Banana Fruity (F) 30 lg/la
Ethyl hexanoate Green apple, fruity Fruity (F) 5 lg/lc
trations of 10, 20, 30, 40 and 50 g/l (P10 to P50). A pollen-free
Ethyl octanoate Pear, pineapple Fruity (F) 5 lg/la sample (sample B) was also included. Once enriched diluted honey,
Diethyl succinate Fruity Fuity (F) 1200 lg/lb the turbidity and yeast-assimilable nitrogen (YAN) contents of the
2-phenylethanol Roses, lilac Floral (FL) 10000 lg/lc honey must were determined (Table 2).
Hexanoic acid Plant, herbs Green (G) 420 lg/la
The honey must was inoculated with a commercial wine yeast
1-hexanol Herbs, cut herbs Green (G) 1100 lg/lb
Phenylacetic acid Honey, pollen Honey (H) 10000 lg/le strain of S. cerevisiae, ENSIS-LE5Ò (Tensum Tecnologicas, Barcelona,
Phenylacetaldehyde Honey, beeswax Honey (H) 5 lg/lf Spain), at a dose of 15 g/hl and incubated at 25 °C.
Phenylethyl acetate Honey, sweet Honey (H) 250 lg/ld Throughout the fermentation process, the density and biomass
Benzaldehyde Bitter almond Sweet (S) 2000 lg/lb concentrations of the must were routinely measured. Before each
Buthyl lactate Creamy, milky, Sweet (S) 1000 lg/lb
sweet
determination, the tanks were homogenised with a charge/dis-
Ethyl lactate Fruity, buttery Other (O) 15000 lg/lb charge of honey must. The sample was centrifuged (10000 rpm,
Ethyl acetate Solvent Other (O) 12 lg/le 10 min), and the density was determined using an electronic
Isoamyl alcohols Solvent, alcohol Other (O) 30 lg/lf DMA 48 densimeter (Anton-PaarÒ, Net IterLab. Salt, Madrid, Spain).
Acetaldehyde Bitter almond, Other (O) 10 g/lc
The yeast cell biomass was determined by a staining method; the
pineapple
sample was stained with methylene blue to distinguish between
a
Escudero, Hernández-Orte, Cacho, and Ferreira (2000). live and dead cells, and examined under a light microscope in a
b
Peinado et al. (2004).
c hemocytometer (Neubauer counting chamber) (Paiting & Kirsop,
Mayén et al. (2005).
d
Selli, Cabaroglu, Canbas, Erten, and Nurgel (2003). 1990).
e
Gómez-Míguez, Gómez-Míguez, Vicario, and Heredia (2007). Once the fermentations had finished (the density measure was
f
Ferreira, Ortín, Escudero, López, and Cacho (2002). unchanged and the residual sugar <5 g/l), the meads were cold
576 A. Roldán et al. / Food Chemistry 126 (2011) 574–582
Table 2
Pollen influence on physicochemical characteristics of honey musts (mean ± SD, n = 3).
B P10
Bè 11.1 ± 0.1 11.9 ± 0.2
TA (g TH2/l) 1.01 ± 0.09 1.20 ± 0.10
pH 3.60 ± 0.01 3.62 ± 0.02
Abs 280 nm 2.57 ± 0.32 3.88 ± 0.57
Abs 420 nm 0.17 ± 0.09 0.29 ± 0.10
Turbidity (NTU) 137 ± 10 2377 ± 150
YAN (mg/l) 53.2 ± 5.0 70.0 ± 8.5
B, control must honey; P10–P50, must honey with pollen from 10 g/l until 50 g/l; TA, total acidity in g of tartaric acid/l; abs, absorbance at 280 and 420 nm; YAN: yeast-
assimilable nitrogen.
(6 °C) for one week, and subsequently treated with gelatine (4 g/hl)
and bentonite (40 g/hl). Finally, the meads were filtered and bot-
tled. All trials were triplicated to ensure statistical significance.
2.2. Nitrogen, protein and amino acid analysis of honey and pollen
The total nitrogen was determined with the Kjeldahl method
adapted for the Velp Scientifica digestion (model DK6) and distilla-
tion (model UDK127) units (Velp Scientifica s.r.l., Milano, Italy).
The total protein was calculated by multiplying the honey and pol-
len nitrogen content by 6.25.
Amino acids were isolated from commercial honey and pollen
according to the method described by González et al. (2006). The
derivatization mixture was prepared by dissolving 50 mg of re-
agent OPA (phthaldialdehyde) in 2.5 ml of ethanol, 25 ll 2-
mercaptoethanol (both reagents from Sigma–Aldrich Química
S.A., Madrid, Spain), and 4.45 ml of borate buffer (0.4 M, pH 9.5).
The derivatization procedure was as follows: 50 ll borate buffer
0.4 M (pH 9.5), 60 ll OPA reactive and 1 ml of isolated amino acids.
The borate buffer and OPA reagent were added by the autosam-
pler/dilutor automatic injector at 1 min before the injection. Direct
injection of derivatised samples was made on a Gilson, Inc., liquid
chromatograph (322 Pump equipment and UV/VIS detector model
151) fitted with a SUPELCOSIL™ LC-18 column (3 lm,
150  4.6 mm) set at room temperature. The chromatographic
conditions were as follows: 0.8 ml/min; volume injection 20 ll;
and solvents A (sodium acetate buffer (50 mM, pH 6.8): methanol
(9:1) with 2% of tetrahydrofurane) and B (methanol with 0.5% of
tetrahydrofurane). The gradient consisted of 85% A, 72% A
(3 min), 56% A (25 min), 44% A (35 min) and 20% A (45 min). Fluo-
rimetric detection was carried out using excitation and emission
wavelengths of 324 and 420 nm, respectively.
Identification and quantification was achieved through reten-
tion times obtained from pure compounds and the calibration
curves of a kit of high purity L-amino acids (Sigma–Aldrich),
respectively.
2.3. Analysis of honey must and meads
Several measurements of the honey musts were taken: Beaumé
degree, pH, total acidity, sulphurous anhydride and yeast-assimila-
ble nitrogen contents (YAN). Alcohol content, total acidity (TA),
volatile acidity (VA), pH, residual sugars (RS), absorbance at
280 nm and 420 nm and major and minor aromatic compounds
were determined in the meads.
The Beaumé degree was determined by areometry, using Dujar-
din-Salleron areometers (Laboratoires Dujardin-Salleron, France).
The pH was determined by measuring the samples directly, using
a digital micro-pHmeter CRISON-2001Ò (Crison Instruments Corp.,
Barcelona, Spain) with automatic temperature compensation. The
turbidity was determined by nephelometry (NTU) with a
2100ANÒ equipment (Hach Company, Loveland, USA). The quantity
P20 P30 P40 P50
12.1 ± 0.2 12.5 ± 0.1 12.7 ± 0.1 12.9 ± 0.1
1.46 ± 0.19 1.78 ± 0.11 2.00 ± 0.11 2.23 ± 0.12
3.64 ± 0.02 3.55 ± 0.01 3.60 ± 0.02 3.66 ± 0.01
5.18 ± 0.71 6.78 ± 0.43 7.65 ± 0.52 8.72 ± 0.39
0.35 ± 0.05 0.42 ± 0.04 0.51 ± 0.07 0.61 ± 0.11
3740 ± 200 5367 ± 180 8688 ± 175 >10000
92.4 ± 6.3 120.4 ± 6.1 140.0 ± 7.2 168.0 ± 5.7
of yeast-assimilable nitrogen (YAN) was determined according to
the abbreviated formol index method proposed by Aerny (1997).
The alcohol content, total acidity and volatile acidity were
determined according to the official methods of wine analysis
(OIV (Office International de la Vigne et du Vin), 1990). Residual
sugars were measured using an enzymatic test with ViniTestÒ
equipment (VinoBios, Denmark). Absorbance was measured with
a Genesys 10UV spectrophotometer (Thermo, USA). Absorbance
at 280 nm indicated the total phenol index (TPI).
2.4. Analysis of volatile compounds
Major volatile compounds (acetaldehyde, ethyl acetate, metha-
nol, 2-methylbutanol and 3-methylbutanol) were analysed using a
GC equipped with an FID detector (HP 5890 Series II) on a Carbo-
wax 20 M column (50 m, 0.25 mm ID, 0.25 lm). The injector and
detector temperatures were 175 and 225 °C, respectively. The car-
rier gas was hydrogen (1 ml/min). The oven temperature was 35 °C
for the first 5 min, with a ramp of 5 °C/min until the temperature
reached 100 °C. A direct injection of 20 ll of distilled sample was
made. For identification and quantification of major volatile com-
pounds, 4-methyl-2-pentenol was added as internal standard and
pure standards compounds (Sigma–Aldrich Química, S.A., Madrid,
Spain) were used to determine the retention times and calibration.
Minor volatile compounds were determined by GC–MS, after a
solid phase extraction (SPE) following the method described by
Di Stefano (1991). A GC–MS model VoyagerÒ (Termoquest, Milan,
Italy) was used with a Supelcowax-10 column (L 60 m, ID
0.32 mm, PD 0.5 lm). The operation conditions of the GC were
the following: injector and detector temperature, 250 °C; oven
temperature of 40 °C for 5 min, followed by a ramp of 2 °C/min,
and 200 °C for 5 min); 2 ll of sample volume in splitless mode
(40 s); and carrier gas, He (1 ml/min). The MS conditions were as
follows: electronic impact mode (EI+) at 70 eV; origin temperature,
220 °C; interface temperature, 320 °C; scan index at 1 scan/s; and
mass acquisition range of 45–400 amu.
Identification of the minor volatile compounds was performed
by GC–MS retention indices (authentic chemicals) and mass spec-
tra comparisons (authentic chemicals and Xcalibur spectral library
collection). All the volatile compounds standards used for the iden-
tification and quantification were obtained from Sigma–Aldrich
(Sigma–Aldrich Química, S.A. (Madrid, Spain). Semiquantitative
analyses were carried out, assuming a response factor equal to one.
2.5. Analysis of odour activity values (OAVS)
To simplify the aromatic profile, odour activity values (OAVs)
were calculated as the ratio between the concentration of each
compound and its perception threshold (Table 1). Only compounds
with OAVs greater than 1 were considered contributors to the
meads’aromatic profiles. The volatile compounds were grouped
in odour series according to similar aroma descriptors in order to
 A. Roldán et al. / Food Chemistry 126 (2011) 574–582 577

relate the quantitative information of the chemical analysis with
sensorial perceptions, resulting in a more objective approach (Pei-
nado, Moreno, Bueno, Moreno, & Mauricio, 2004). An odour series,
depending of their main odour descriptor, was assigned to each
component. The following aromatic series were chosen as descrip-
tors for meads: green (G), fruity (F), floral (FL), oxidation (DO),
sweet (S), honey (H) and others (O). The sum of the OAVs obtained
for each aroma descriptor was calculated.
2.6. Sensory evaluation
The sensory evaluation of meads was performed by a panel of
ten panellists, ages 30–50 ages (four female and six male) with
wine tasting experience, and trained in the evaluation of meads.
Sensory analysis was performed in individual booths with con-
trolled illumination located in the tasting room of the Andaluz
Center of Wine Research (CAIV, Puerto Real, Cádiz).
The meads were presented in standard tasting glasses 3591 (ISO
3591, 1997) and covered with watch glass to minimise the evapo-
ration of volatile compounds. The tastings were conducted be-
tween 11:30 am and 14:00 pm and the meads were at a
temperature of 10–12 °C. Each taster was given specific tasting
notes to evaluate the visual (turbidity and colour), aroma (quality
and intensity) and taste (quality and intensity) characters, as well
as the general acceptability of the product. Each character was
scored from 0 to 5 according to the increasing intensity. Some as-
pects of interest such as characteristic odour (honey, fruity, flower)
and defects odour (rancid, oxidised, musty) were also considered.
2.7. Statistical analysis
Means and standard deviations were calculated and significant
differences were evaluated using Student’s t-test. Statistical pro-
cessing was carried out using the STATISTICA Release 7 (Statsoft,
Inc.-USA) statistical package.
3. Results and discussion
3.1. Effects of pollen addition on honey must composition
closely with the pollen concentration (YAN: R2 = 0.9951; turbidity:
R2 = 0.9847 for YAN).
YAN constitutes of ammoniacal nitrogen, amino acids, small
peptides and nitrogen that can be easily assimilated and is essen-
tial for yeast growth (Alexandre & Charpentier, 1998; Bell & Hens-
chke, 2008; Bisson & Butzke, 2000) and for the complete
development of fermentation (Bisson, 1999; Bisson & Butzke,
2000). Pollen is a major source of YAN and could be used to enrich
nitrogen-poor media. The nitrogen and protein contents of pollen
were 2.52% (±0.60) and 15.74% (±0.60), respectively, while the
nitrogen and protein contents of honey were 0.10% (±0.07) and
0.63% (±0.07), respectively. As expected, the honey that was used
for our mead exhibited a low YAN concentration, and activators
were necessary to optimally ferment the honey must.
The total amino acid contents of the pollen and honey were
15.1% and 0.70%, respectively (Table 3). The proportions of free
amino acids were approximately the same in both samples:
14.2% of the total amino acid content for pollen and 14.3% for hon-
ey. Honey and pollen amino acids were mainly proteins and pep-
tides. Pollen had a higher amino acid content than honey;
however, the pollen/honey ratio (P/H) was very similar for the dif-
ferent amino acids measured, which indicates that both substrates
are closely correlated and both have the same origin. The pollen
content of honey was indicated by its amino acid proportions. Con-
sequently, pollen addition provides a good supply of YAN, and the
overall percentage of each amino acid remains constant when pol-
len is added. Moreover, the amino acid content of honey must is
enriched in a balanced way by pollen addition. The main amino
acids in both pollen and honey are proline (52%), glutamic acid
(15%) and phenylalanine (14%), followed by aspartic acid
(3%) and threonine, tryptophan and valine (2%). These results
are in accordance with previous reports that pollen contains 16%
amino acids, with major contributions of proline, glutamic acid,
aspartic acid, lysine and leucine (Human & Nicolson, 2006). Honey
only contains 1% (w/w) amino acids, primarily proline, phenylala-
nine, tyrosine and lysine (Bouseta et al., 1996; Hermosín, Chicón, &
Cabezudo, 2003).
Each 10 g/l of pollen added to a fermentation medium provides
on average 70% of the amino acids that were already present in 1 l
of honey-must. The YAN levels in the P40 and P50 meads were
above 140 mg/l, the minimum amount necessary to complete the

The diluted honey had a density of 11.1 °Be (188.7 g/l of sugars),
a turbidity of 137 NTU, a pH of 4.42 and total acidity of 0.7 g tar- Table 3
taric acid/l. After pollen was added in different concentrations Distribution of total and free aminoacids (as mg/g sample) for commercial honey and
(0–50 g/l), the Baumè degree, total acidity, pH, total polyphenol in- pollen, and pollen/honey (P/H) ratio (mean ± SD, n = 4).
dex (TPI, absorbance 280 nm), turbidity and yeast-assimilable Total aminoacids Free aminoacids
nitrogen (YAN) of each sample were measured (Table 2). This table
Honey Pollen P/H Honey Pollen P/H
shows the change in the Baumè degree with pollen addition (R2
Ala 0.07 ± 0.01 1.49 ± 0.49 21.6 nq 0.02 ± 0.00 –
equal to 0.9847), which corresponded to approximately 0.4 g of su-
Arg 0.12 ± 0.02 3.46 ± 0.76 30.1 0.02 ± 0.01 0.31 ± 0.07 18.2
gar per gram of pollen added. Therefore, pollen addition contrib- Asp 0.23 ± 0.05 4.78 ± 1.33 21.0 0.03 ± 0.01 0.69 ± 0.23 20.3
uted to the increased probable alcohol content of the mead. Total Cys 0.03 ± 0.00 0.50 ± 0.05 20.0 nq 0.07 ± 0.01 –
acidity was also enhanced by pollen addition (R2 = 0.9909), with Glu 1.07 ± 0.08 22.81 ± 1.02 21.3 0.16 ± 0.01 2.92 ± 0.10 18.3
a 0.02 g increase in tartaric acid per gram of pollen. However, the Gly 0.04 ± 0.01 0.82 ± 0.31 20.5 0.01 ± 0.00 0.11 ± 0.01 18.3
His 0.02 ± 0.00 0.37 ± 0.04 21.8 nq 0.05 ± 0.00 –
pH values remained constant (mean pH 4.4) indicating that, given
Ileu 0.04 ± 0.01 0.80 ± 0.34 22.2 nq 0.11 ± 0.02 –
the low buffering capacity of the honey must, the increase in the Leu 0.08 ± 0.02 1.63 ± 0.57 21.2 0.01 ± 0.00 0.24 ± 0.02 21.8
total acidity after pollen addition was due to the weak acids. Add- Lys 0.05 ± 0.03 1.18 ± 0.53 22.7 0.01 ± 0.01 0.17 ± 0.03 21.3
ing pollen increased the total polyphenol index by approximately Met 0.02 ± 0.00 0.38 ± 0.07 21.1 nq 0.06 ± 0.01 –
Phe 1.00 ± 0.15 21.80 ± 1.09 21.8 0.15 ± 0.02 3.10 ± 0.14 21.1
0.130 absorbance units per gram of pollen, indicating that certain
Pro 3.74 ± 0.78 78.91 ± 1.63 21.1 0.56 ± 0.02 11.59 ± 0.62 20.7
compounds of pollen (polyphenol substances, mainly flavonoids) Ser 0.07 ± 0.02 1.41 ± 0.32 20.4 0.01 ± 0.01 0.27 ± 0.10 24.5
were solubilised in honey must. The honey must colour was also Thr 0.15 ± 0.01 3.90 ± 0.31 25.8 0.02 ± 0.01 0.45 ± 0.03 20.5
modified by the addition of pollen, as can be observed in the absor- Trp 0.16 ± 0.00 3.09 ± 0.10 19.3 0.02 ± 0.01 0.42 ± 0.15 20.0
bance, at 420 nm, values (0.09 absorbance units per gram of pol- Tyr 0.04 ± 0.00 0.75 ± 0.17 21.4 nq 0.18 ± 0.05 –
Val 0.14 ± 0.02 2.93 ± 0.86 20.9 0.02 ± 0.01 0.44 ± 0.11 21.0
len). This increase in absorbance values resulted in more yellow
Total 7.07 ± 0.83 151.0 ± 2.43 21.4 1.02 ± 0.03 21.40 ± 0.09 20.9
and brown honey musts. The turbidity and YAN levels also in-
creased with the addition of pollen, and both measures correlated Nq, no quantified (<0.01 mg/g sample).
578 A. Roldán et al. / Food Chemistry 126 (2011) 574–582

fermentation in grape must (Barre, Blondin, Feuillat, Sablayrolle, & 18 9

Salmon, 1998; Riberéau-Gayón, Dubordieu, Doneche, & Louvand,
16 8
2000). Based on this data, pollen provides a good complement of
amino acids for the honey-must fermentation process. 14 7
At industrial level, turbidity is usually related to the quantity of 12 6

mg/l · days
nutrients in the raw material; it is the first checkpoint in proper Vm ax
fermentation development. Hidalgo (2003) set the optimal turbid- 10 5

ity values from 50 to 200 NTU for grape must. However, the addi-
tion of pollen, even at lower doses, significantly increases the
turbidity values, exceeding 2000 NTU in all cases.
days
t m ax
8 4
6 3
4 2

3.2. Pollen’s influence on fermentation kinetics
The evolution curves of the relative density during the fermen-
tation of the different honey musts are shown in Fig. 1. The fer-
mentation kinetics were significantly different between the
control must (B) and the honey must with added pollen. In all
cases, fermentation was complete, and all of the produced meads
reached a residual sugar content of less than 0.6 g/l. Without a fer-
mentation activator like pollen (B), the fermentation lasted
approximately 6 weeks, confirming that honey is low in nutrients
for yeast fermentation (Bahiru, Mehari, & Ashenafi, 2001; Gupta
& Sharma, 2009) and that, as Barre et al. (1998) showed, this lack
prolongs the fermentation process. Comparing between the differ-
ent concentrations of pollen, the fermentation rate (Fig. 1) in-
creased with pollen addition. Fig. 2 shows the maximum
fermentation rate (Vmax) and the time required to reach the maxi-
mum rate (tmax) for the different concentrations of added pollen.
The control mead (0 g/l added) showed a significantly lower fer-
mentation rate and a higher time to reach the maximum rate than
the honey musts with added pollen. An increase in fermentation
rate (y = 0.0074x2 + 0.6017x + 3.9225, R2 = 0.9916) and reduction
of the time to maximum with increasing concentrations of pollen
were observed. The honey musts containing more than 30 g/l of
added pollen showed the highest fermentation rate and the
shortest maximum time. The increase in the fermentation rate
was correlated with increase in YAN and turbidity values, and
resulted in improving the fermentation kinetics (Alexandre &
Charpentier, 1998; Bisson & Butzke, 2000). These findings corre-
spond with the results of Vilanova et al. (2007), which showed that
the fermentation rates increased in response to increased nitrogen
concentrations in chemically defined fermentation media. More-
over, the high levels of polyunsaturated fatty acids found in pollen,
such as linoleic and linolenic acids (Xu, Sun, Dong, & Zhang, 2009),
must be taken into account because these are metabolised by the
1,005
0,995
0,985
0,975
Density (g/ml)
2 1
0 0
0 10 20 30 40 50 60
Pollen concentration (g/l)
Fig. 2. Maximum fermentation rate (Vmax in mg/l
day) and time to reach the
maximum rate (tmax in days) reaching with pollen addition.
yeast cells during fermentation (Sroka & Tuszýnski, 2007) and
may improve the fermentation kinetics.
To study the effect of pollen addition on the alcoholic fermenta-
tion yield and efficiency, the yield [produced alcohol (% w/v)/con-
sumed sugars (% w/v)  100] and efficiency [(produced alcohol/
theoretical alcohol from consumed sugars)  100] were calculated.
The results (Table 4) show that the diluted honey’s alcoholic fer-
mentation yield (B) was about 37 g ethanol/100 g fermentable sug-
ars, representing an alcoholic fermentation efficiency of about 81%.
The addition of pollen improved the fermentation yield and
efficiency by more than 7% and 10%, respectively.
3.3. Pollen’s influence on the physicochemical characteristics of meads
The physicochemical analyses of the final meads are shown in
Table 5. The final characteristics depended on the alcoholic fer-
mentation and the amount of pollen added to the honey must.
The total acidity increased and the pH decreased during alco-
holic fermentation. These results confirmed those of other authors
(Sroka & Tuszýnski, 2007), who affirm that as honey must fer-
ments, succinic and acetic acids are formed. These acids increase
the meads’ total acidity and reduce their pH. As can be observed
in Table 5, the meads’ total acidity decreased with pollen addition,
which could be primarily due to a small deviation for the yeast’s
metabolism toward organic acid production, principally acetic
acid. In addition, pollen provides potassium and calcium salts
(Ouchemoukh et al., 2007; Silva et al., 2009) that may lead to sali-
B
nization, resulting in decreased acidity. As a result of this decrease
P10
in acidity, the final mead pH increased to more than 4.0 with pol-
P20
len addition. These high pH values, especially for additions of 40
P30
P40

0,965 P50

0,955 Table 4
Alcoholic fermentation yield and efficiency in meads (mean ± SD,
0,945
n = 3).
0,935
Yield (%) Efficiency (%)
0,925
B 36.7 ± 0.5 81.4 ± 0.8
0,915 P10 44.5 ± 0.2 98.7 ± 0.5
P20 44.0 ± 0.3 97.5 ± 0.3
0,905
P30 43.4 ± 0.5 96.2 ± 0.4
0 10 20 30 40 50
P40 43.0 ± 0.4 95.2 ± 0.2
time (days) P50 43.3 ± 0.8 90.7 ± 0.5

Fig. 1. Relative density evolution of the honey-musts during fermentation. (B) B, control must honey; P10–P50, must honey with pollen from
Control honey must.; P10–P50: honey musts with pollen from 10 g/l to 50 g/l. 10 g/l until to 50 g/l.
 A. Roldán et al. / Food Chemistry 126 (2011) 574–582 579

Table 5
Pollen influence on physicochemical characteristics of meads. (mean ± SD, n = 3).

B P10 P20 P30 P40 P50

EtOH (% v/v) 9.04 ± 0.02 11.74 ± 0.06 11.80 ± 0.10 12.03 ± 0.08 12.09 ± 0.05 12.39 ± 0.12
pH 2.95 ± 0.15 3.59 ± 0.09 3.78 ± 0.11 3.94 ± 0.07 4.04 ± 0.06 4.05 ± 0.05
TA (g TH2/l) 4.87 ± 0.21 4.05 ± 0.17 3.40 ± 0.08 3.29 ± 0.07 3.15 ± 0.10 3.37 ± 0.09
VA (g AcH/l) 1.370 ± 0.179 0.477 ± 0.029 0.515 ± 0.065 0.481 ± 0.014 0.433 ± 0.077 0.710 ± 0.104
Abs 280 nm 4.22 ± 0.20 5.54 ± 0.23 7.71 ± 0.54 10.62 ± 0.31 13.29 ± 0.13 19.02 ± 0.27
Abs 420 nm 0.080 ± 0.002 0.152 ± 0.011 0.205 ± 0.005 0.284 ± 0.024 0.327 ± 0.008 0.572 ± 0.083
RS (g/l) <0.6 <0.6 <0.6 <0.6 <0.6 <0.6

B, control must honey; P10–P50, must honey with pollen from 10 g/l until to 50 g/l; TA, total acidity in g of tartaric acid/l; VA, volatile acidity in g of acetic acid/l; abs,
absorbance at 280 and 420 nm; RS, residual sugars.

and 50 g/l of pollen (P40 and P50), might reduce the microbial
stability of meads and could justify correcting their pH after
fermentation.
The volatile acidity was elevated in all cases, reaching levels of
almost 0.5 g/l. Moreover, the control mead showed acetic acid lev-
els above the sensory threshold for table wines (0.7 g AcH/L). Low
amounts of YAN favour the production of acetic acid (Vilanova
et al., 2007), and yeast metabolism may benefit by the accumula-
tion of acetic acid as an intermediate secondary product (Barre
et al., 1998) during a slow fermentation.
Mead colour, generally measured at 420 nm (a yellow hue), in-
creased with the increasing content of pollen, as did its absorbance
at 280 nm, indicating its total phenol index (TPI). Phenolic com-
pounds are naturally present in honey and pollen but vary widely
depending on the floral origin (Nagai et al., 2006; Ouchemoukh,
Louaileche, & Schweitzer, 2007). Polyphenols are of great impor-
tance to the colour, astringency and antioxidant properties, which
are highly correlated (Paixão, Perestrelo, Marques, & Câmara,
2007). A significant increase in the meads’ absorbance at 280 nm
values (PTI) was observed after alcoholic fermentation. In the hon-
ey musts, this increase was linearly correlated with the amount of
pollen added; however, in the meads, this increase was exponen-
tial (y = 4.1933e0.0299x, R2 = 0.9976). This nonlinear behaviour could
be due to the proportion of alcohol present in the mead. As can be
seen in Table 5, the TPI value increased with the alcoholic level
(y = 0.049x + 11.46, R2 = 0.9835). Therefore, alcohol favours the
extractability of compounds that absorb at 280 nm, such as poly-
phenols, flavonoids, phospholipids, and proteins. Ethanol could ex-
pand the outer layer of the pollen wall (exine) and improve the
solubilisation of its bioactive compounds.
The residual sugars were below the detection limit of the Vini-
TestÒ equipment (<0.6 g/l) for all meads. This result indicates that
all honey-musts sugars were converted to ethanol and/or other
secondary metabolites of fermentation.
It appears that using pollen as a fermentation activator not only
improves the fermentation kinetics, but also contributes to im-
prove the meads’ final characteristics.
3.4. Pollen’s influence on aromatic characteristics of meads
Secondary metabolites like aromas are generally produced by
yeasts (Fleet, 2003). Their production is influenced by the yeast
strain used and the compounds that provide the essential nutrients
for yeast metabolism and serve as precursors for the final aromas.
The results for the major aromatic compounds are shown in
Table 6. The higher alcohols contributed most to the major volatile
compounds. In general, isoamyl alcohols increased with the addi-
tion of pollen, which agrees with our previous experiments related
to honey fermentation (Roldán et al., 2008; Vidrih & Hribar, 2007).
The sensory threshold value of isoamyl alcohols is 300 mg/l; how-
ever, they produce an unpleasant flavour and taste in wines in con-
centrations higher than 400 mg/l (Gil, Cabellos, Arroyo & Prodanov,
2006). The concentrations in the meads did not exceed this limit.
Isoamyl alcohols are derived from amino acids, and pollen compo-
nents appear to contribute to their formation. The methanol con-
tent in the control samples was low, but it increased with higher
pollen concentrations. This increase might be due to the large
amount of pectins in pollen and the enzymatic hydrolysis during
fermentation that results in methanol. The methanol levels corre-
lated with the amount of pollen added (y = 0.4616x + 6.609,
R2 = 0.9904), indicating that the methanol content was not derived
from the yeast’s activity during fermentation. Moreover, the acet-
aldehyde concentrations were the lowest in the control mead
and higher for the rest of meads. The ethyl acetate values were re-
lated to the acetic acid content obtained during fermentation, and
were expressed as volatile acidity. The mead with higher values of
volatile acidity also had higher levels of ethyl acetate.
Regarding the total minor volatile content (Table 7), pollen
addition increased the control value by 266–310%. P30 had the
least increase in minor volatile content (166%) and P10 and P50
had the highest (210%). There was no correlation between the
amount of added pollen and the total minor volatiles content.
The substrates necessary for the production of these volatiles were
obtained even with low concentrations of pollen (10 g/l). The main
contributors to the total minor volatiles content in meads were

Table 6
Major aromatic compounds of the obtained meads (mg/l). (mean ± SD, n = 3).

Compound B P10 P20 P30 P40 P50 OS

Isoamyl alcohols
2-methyl butanol 24.11 ± 0.16 61.55 ± 0.41 73.74 ± 0.41 71.55 ± 0.48 78.97 ± 0.53 83.50 ± 0.56 O
3-methyl butanol 91.04 ± 0.92 187.22 ± 1.88 218.86 ± 1.88 211.08 ± 2.12 234.82 ± 2.36 234.10 ± 2.36 O
Average total isoamyl alcohols 115.15 ± 1.06 248.78 ± 2.29 292.60 ± 2.70 282.62 ± 2.61 313.79 ± 2.89 317.60 ± 2.93
Others
Methanol 5.66 ± 0.64 11.80 ± 2.00 17.13 ± 1.93 19.91 ± 2.25 25.54 ± 2.88 29.05 ± 3.28
Acetaldehyde 8.01 ± 0.37 19.08 ± 0.91 18.11 ± 0.85 22.15 ± 1.04 27.61 ± 1.29 30.18 ± 0.62 O
Ethyl acetate 21.80 ± 12.98 13.15 ± 7.83 15.33 ± 9.13 8.75 ± 5.21 13.68 ± 8.15 19.25 ± 11.46 O
Average total others 35.47 ± 5.02 43.03 ± 10.91 50.57 ± 7.16 50.81 ± 7.19 66.83 ± 9.46 78.48 ± 8.70
Total major aromas 150.63 ± 2.41 291.81 ± 5.22 343.17 ± 5.49 333.43 ± 5.34 380.62 ± 6.09 396.08 ± 6.07

B, control must honey; P10–P50, must honey with pollen from 10 g/l until to 50 g/l; OS, odour series of the corresponding chemical compound. G, green odour; F, fruity odour;
DO, oxidation odour; O, other odour; S, sweet odour; FL, floral odour; H, honey odour.
580 A. Roldán et al. / Food Chemistry 126 (2011) 574–582

Table 7
Minor aromatic compounds of the obtained meads (lg/l). (mean ± SD, n = 3).

Compound B P10 P20 P30 P40 P50 OS

Acids
Isovaleric acid 109.14 ± 30.05 572.09 ± 157.53 325.69 ± 89.68 396.04 ± 109.05 493.42 ± 135.87 614.81 ± 169.29 DO
Hexanoic acid 124.66 ± 20.77 903.12 ± 150.46 1285.48 ± 214.16 1067.85 ± 177.85 1195.46 ± 199.16 1337.90 ± 222.90 G
Octanoic acid 133.50 ± 13.06 1840.15 ± 180.04 2137.20 ± 209.10 1452.43 ± 142.11 1408.94 ± 137.85 1589.03 ± 155.47 DO
Phenylacetic acid 96.79 ± 42.47 204.59 ± 89.77 268.12 ± 117.64 151.31 ± 66.39 209.56 ± 91.95 205.12 ± 90.00 H
Average total acids 464.09 ± 78.10 3519.95 ± 592.39 4016.49 ± 675.96 3067.62 ± 516.22 3307.38 ± 556.62 3746.86 ± 630.58
Percentage of total 8.2 20.0 23.1 20.3 21.1 21.3
Esters
Isoamyl acetate 2.12 ± 0.90 61.83 ± 26.30 64.06 ± 27.25 58.46 ± 24.87 73.17 ± 31.12 77.62 ± 33.01 F
Ethyl hexanoate 0.74 ± 0.64 22.50 ± 19.53 33.95 ± 29.49 32.23 ± 27.99 33.09 ± 28.73 36.03 ± 31.29 F
Ethyl lactate 14.01 ± 6.23 54.36 ± 24.18 38.52 ± 17.14 42.85 ± 19.06 15.59 ± 6.94 627.34 ± 279.11 F
Ethyl octanoate ND 80.40 ± 27.41 113.56 ± 36.52 99.00 ± 12.03 81.58 ± 23.78 72.40 ± 35.41 F
Buthyl lactate ND ND ND 1.22 ± 0.54 1.70 ± 0.38 45.71 ± 20.68 F
Ethyl 3- hidroxy butanoate 4.72 ± 1.02 25.45 ± 5.51 26.41 ± 5.72 37.26 ± 8.07 42.06 ± 9.11 32.27 ± 6.99 DO
Diethyl succinate 54.55 ± 7.32 168.63 ± 22.64 218.95 ± 29.39 187.85 ± 25.22 220.80 ± 29.64 702.39 ± 94.29 F
Phenylethyl acetate 18.92 ± 0.22 226.47 ± 2.67 205.05 ± 2.41 250.09 ± 2.94 306.20 ± 3.60 128.64 ± 1.51 F
Average total esters 95.06 ± 18.25 639.64 ± 122.80 700.49 ± 134.49 708.96 ± 136.11 774.19 ± 148.64 1722.40 ± 330.69
Percentage of total 1.7 3.6 4.0 4.7 4.9 9.8

B, control must honey; P10–P50, must honey with pollen from 10 g/l until to 50 g/l; OS: odour series of the corresponding chemical compound. G, green odour; F, fruity
odour; DO, oxidation odour; O, other odour; S, sweet odour; FL, floral odour; H, honey odour.

alcohols (>60%) and acids. Adding pollen increased the alcohol con-
tent, particularly 2-phenylethanol. However, their contribution to 80
the overall volatile was 20% lower than for the control (Table 7). GREEN (G)
FRUITY (F)
The contribution of the remaining aromatic families to the total ar- OXIDATION (DO)
oma increased with increased pollen content. Pollen addition sig- 70
HONEY (H)

nificantly increased the proportion of acids in the meads’ aroma,

30
particularly octanoic and hexanoic acids. Acids were increased to
levels up to 500% greater than those of the control mead, but there
was no correlation between the production of these acids and the
pollen content of the honey must. 20
ΣOAV

The meads with added pollen showed an increase in the propor-

tion of esters, especially ethyl succinate and phenylethyl acetate. In
general, fruity odour was noted, and ethyl lactate and butyl lactate
levels were high in P50. High butyl lactate levels in P50 could be
due to the lactic acid derived from fermentation by heterolactic
bacteria after alcoholic fermentation. These values could also jus-
tify the high levels of ethyl acetate and volatile acidity of this
mead. The need to correct the pH value and the sulphur content
in this mead once the fermentation is complete could also be
justified.
As can be seen in Table 7, aldehydes, particularly phenylacetal-
dehyde in P50, followed a trend similar to that of the esters. In gen-
eral, the production of these volatile compounds in the studied
meads was related to the addition of pollen. No significant relation-
ship with the amount of pollen added was observed. Some aromas
that increased with pollen addition, such as 2-phenylethanol, benz-
aldehyde, phenylacetaldehyde and phenylacetic acid, are naturally
present in honey, independent of its geographical or botanical ori-
gin (Bouseta et al., 1996; Fleet, 2003; Alissandrakis, Tarantilis,
Harizanis & Polissiou, 2007). These compounds are usually related
to the aromas of honey, pollen, and flowers and to the waxy charac-
ter of honeys (Castro-Vázquez, Díaz-Maroto, & Pérez-Coello, 2007).
The increase of these compounds suggests that some pollen compo-
nents, such as phenylalanine, are precursors to their formation.
Attending to the odour activity values (OAVs), as can be ob-
served in Fig. 3, in general, pollen addition was associated with in-
creased values for all of the aromatic groups, especially for the
fruity and oxidation groups, regardless of the amount of pollen
added. Higher concentrations of pollen significantly increased the
aromas in the honey series, mainly associated to high levels of phe-
nylacetaldehyde. The other aromatic series followed a different
trend, reaching a maximum and a minimum for fruity and oxida-
tion series, respectively, with the added pollen dose.
10
0
B P10 P20 P30 P40 P50
Pollen concentration (g/l)
Fig. 3. Influence of pollen addition on the OAVs of meads. (B) Control must honey;
P10–P50: must honey with pollen from 10 g/l to 50 g/l; G: green odour; F: fruity
odour; DO: oxidation odour; H: honey odour.
Therefore, the combination of increased nutrient level and a
slower fermentation process might have favoured an increase in
the total aroma content, as well as a different balance of aromas.
P20 and P30 showed the main contribution of the fruity series
and the lowest of the oxidation series. The principal contributor
to the fruity series was ethyl octanoate; octanoic acid was the main
contributor to the oxidation series.
3.5. Pollen’s influence on sensory characteristics of meads
Along with the product’s aromatic profile (OAVs), tasting is
indispensable in evaluating the organoleptic characteristics of
meads. A more complete profile is obtained by including visual,
gustatory and tactile aspects along with aromas.
The tasting test involved rating the following parameters along
a 10-point scale: turbidity, colour, aroma quality and intensity,
taste quality and intensity, and acceptability. The tasting commit-
tee members could also write down any additional aroma or
 A. Roldán et al. / Food Chemistry 126 (2011) 574–582
(p<0,001)
Acceptability
10
8 criteria, the dose of 30 g/hl of pollen is the most appropriate
Taste intensity
6
Turbidity
(p>0,05) (p>0,05)
4 B
2 P10
P20
0
P30
Taste quality Color P40
P50
(p<0,001) (p<0,001)
Aroma intensity Aroma quality
(p<0,01) (p<0,001)
Fig. 4. Effect of pollen additon on sensorial evaluation of meads. (B) Control must
honey; P10–P50: must honey with pollen from 10 g/l to 50 g/l.
flavour characteristics they observed. The sensory analysis results
are presented in Fig. 4.
The ‘‘p’’ values shown in the figure indicate whether the differ-
ence presented by each parameter evaluated, at different pollen
addition, compared with control mead was very significant
(p < 0.001), significant (0.001 < p < 0.01), insignificant (0.01 < p <
0.05) or nonsignificant (p > 0.05).
The most significant differences between the added-pollen
meads and the control mead were in colour, aroma quality, taste
quality and acceptability. The other parameters were not signifi-
cantly different. The meads judged most acceptable (P30 and
P40) had the highest aroma and flavour quality ratings. Samples
P10, P20 and P50 were comparably evaluated. The major difference
between them was the colour value, which increased linearly
(from pale yellow to amber) with pollen addition.
The aroma quality appeared to be related to the sum of OAVs
(Fig. 3). The control’s aroma was described as floral (associated
with phenylethanol) and vinegar-like acid masking other aromas,
which decreased the aroma quality, as was confirmed by the pres-
ence of isovaleric and hexanoic acids and by the sample’s elevated
total and volatile acidity (Table 5). Ethyl acetate (a solvent-like
odour) was also noted in the control. P10 was described as ‘‘medic-
inal’’ and ‘‘floral’’, while the P20 and P30 meads evoked almonds,
dried fruits, apple, caramel and sweets. P20 was also described as
smelling of pineapple associated with acetaldehyde, which was
significantly higher for this mead. P40 differed from the other sam-
ples mainly due to its profound honey aroma. P50 was character-
ised by toasted, bitter almond and honey scents that masked all
other aromas, principally consistent with its high phenylacetalde-
hyde levels.
The pollen content changed the mouthfeel of the meads by
increasing the aromatic body of the sample, which was also stated
by Vidrih and Hribar (2007). Astringency was noted for various
meads, probably due to the increase in polyphenols. Flavour
assessment resulted in deviations similar to those for the aroma
parameters. Thus, meads P40 and P50 were highlighted for sweet-
ness, while the control and P30 were described as sour. A slight bit-
terness was noted for P20. Honey flavour and a floral flavour were
indicated for P40 and P50. The control’s taste was rated signifi-
cantly lower (2.9) than the others due to its sourness and lack of
desirable flavours. Finally, the relative low taste value for P50
(5.1) was attributable to an excessive honey flavour.
4. Conclusions
According to the obtained results it can be concluded that
pollen could be a good and appropriate activator of alcoholic
581
fermentation in the production of mead. The addition of pollen
improves the fermentation kinetics and physicochemical and orga-
noleptic characteristics of the mead. In response to the sensory
because, besides producing mead of better acceptability, it would
mean less use of pollen at industrial level.
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