molecules

Article
Fatty Acids, Volatile and Sensory Profile of Multigrain
Biscuits Enriched with Spent Malt Rootles
Maria Simona Chiş 1 , Anamaria Pop 1, * , Adriana Păucean 1 , Sonia Ancut, a Socaci 2 ,
Ersilia Alexa 3 , Simona Maria Man 1 , Monica Bota 1 and Sevastiţa Muste 1
1 Department of Food Engineering, Faculty of Food Science and Technology, University of Agricultural
Sciences and Veterinary Medicine of Cluj-Napoca, 3–5, Manastur Street, 400372 Cluj-Napoca, Romania;
[email protected] (M.S.C.); [email protected] (A.P.);
[email protected] (S.M.M.); [email protected] (M.B.); [email protected] (S.M.)
2 Department of Food Science, Faculty of Food Science and Technology, University of Agricultural Sciences
and Veterinary Medicine of Cluj-Napoca, 3–5, Manastur Street, 400372 Cluj-Napoca, Romania;
[email protected]
3 Department of Food Control, Faculty of Agro-food Technologies, Banat’s University of Agricultural Sciences
and Veterinary Medicine “King Michael I of Romania”, 119 Calea Aradului Street, 300641 Timişoara,
Romania; [email protected]
* Correspondence: [email protected]; Tel.: +40-744-604728




Received: 19 December 2019; Accepted: 15 January 2020; Published: 21 January 2020


Abstract: Spent malt rootlets, a by-product of the brewing industry, are a rich source of protein,
essential amino acids, healthy fats, polyphenols and minerals, and could be a new promising type
of raw material from the nutritional, economic, sensory, and technical perspectives. However, their
specific aroma profile could limit their addition in baked products. The aim of this work was to study
the effect of spent malt rootlets addition on volatile derivatives of enriched biscuits in relation to
their sensory profile. For this purpose, spent malt rootlets and enriched biscuits (0–25% spent malt
rootlets added) were analyzed by GC-MS techniques, in order to obtain their fatty acids methyl esters
and volatile compounds profile, while for the sensory analysis a nine-point hedonic score test was
used. The results of this study reveal the fatty acids and volatile profile of spent malt rootlets and
of the enriched biscuits with spent malt rootlets pointing out the contribution of fatty acids to the
generation of aroma compounds. The influence of different aroma compounds on the consumer’s
preferences was studied and the optimum level addition of spent malt rootlets in multigrain biscuits
was found to be 15%.

Keywords: biscuits; malt spent rootlets; fatty acids; volatiles; food waste

1. Introduction
Food waste, one of the largest portions of solid waste, has grown to become a global problem.
Every year, nearly 1.3 billion tons of food trash is dumped in landfills or is otherwise disposed of [1].
Its efficient management represents the main objectives of European Union (EU) actions against food
waste and towards sustainable development, aiming to halve food waste per capita at the retail and
consumer level by 2030, and reduce food loss among the food production and supply chains [2].
Nowadays, efforts are being directed towards the exploitation of agro-industrial waste, from both
economic and environmental standpoints. These by-products are recognized as having significant
amounts of biologically active compounds, namely proteins, fibers, polysaccharides, flavor compounds,
or different phytochemicals, while their uncontrolled disposals lead to environmental problems [3,4].
These bioactive compounds can be revalorized as functional ingredients in food, pharmaceutical, health
care, cosmetic, and other products. Using the recovered bioactive molecules as functional ingredients

Molecules 2020, 25, 442; doi:10.3390/molecules25030442 www.mdpi.com/journal/molecules

Molecules 2020, 25, 442 2 of 17

represents a sustainable alternative of food wastes exploitation as an inexpensive source of valuable

compounds, by developing innovative food and non-food products with health-promoting benefits
and at the same time contributing to an efficient waste reduction management [5].
Barley (Hordeum vulgare), a cereal of the Poaceae family, ranks fourth globally in both quantity
produced and cultivation area. It is conventionally utilized for animal fodder (≈66%) or malted for
brewing (≈33%) and food applications (≈2%) [6]. During malting, the sprouted green malt grains
are ‘cleaned’, by removing the protein-rich rootlets, which are primarily discarded or used as animal
fodder. Recently, the incorporation of barley into the human diet has received renewed attention
due to increased scientific evidence showing it to be an excellent source of dietary fiber, particularly
β-glucan [7]. β-Glucan is a soluble fiber that has health benefits, which include decreasing the risk of
heart disease and being effective in lowering blood serum glucose levels [8,9].
Barley malt rootlets, a by-product of the brewing industry, consist of the plumule and radicle of
barley and may include some of the malt hulls [10]. Barley rootlets could contribute to the human
diet through their valuable nutrients such as protein, essential amino acids, healthy fats, polyphenols,
and minerals [7]. Their protein levels are far in excess of those detected in barley or malt grains
31.90–36.75% [7,11]. Waters et al. [7] reported that the ten essential amino acids are heavily represented
in the rootlet portion of malt and are in excess of the wheat flour, whole meal flour, barley and malt
levels for phenylalanine, isoleucine and leucine.
Capitalizing on these agro-industrial wastes into value-added products has recently become a
highly active area of research [12]. Thus, the incorporation, of rootlets into a food product could
increase the protein content of poorer flours and thus increase the net nutritive value of the final baked
goods. Rootlets are considered as an inexpensive source for enriching food due to its composition on
high biological protein value, being first used as a raw material in bakery products in 1997 [13].
The literature survey has clearly shown that spent malt rootlets belongs to the GRAS (generaly
recognised as safe) category and could be a new promising raw material from the nutritional, economic,
sensory and technical perspectives [14–17].
However, changing ingredients may cause changes in reaction precursors and thus may have
an impact on the main reactions occurring during the steps of product-making, such as the Maillard
reaction, caramelization, and lipid oxidation [18].
In this context, this research aimed to study the effect of spent malt rootlets addition on volatile
derivatives of enriched biscuits in relation to their sensory profile. The critical issue in the case
of the addition of spent malt rootlets in food is their sensory profile so it is important to establish
the optimum addition percentage through the sensory analysis in order to correlate the results of
instrumental analysis to human perception. A high quality mix of multigrain (spelt wheat, rice, and
oat), pseudo-cereal (buckwheat), and chia seeds was used in biscuit formulation in order to improve
various nutritional parameters.

2. Results and Discussion

2.1. The Proximate Composition of the Main Raw Materials

The proximate composition of the main raw materials used for the manufacture of the biscuits is
showed in Table 1. The protein and ash contents of SWF (15% and 1.25%, respectively) are close to the
results reported by [19], as well as [20] for whole spelt flour, namely ash (1.95%), protein (15.17%), and
moisture (15.04%). The lipids content is similar with the amount reported by [21] ranging between
3.24–3.5%. The small differences between the results could be due to the environmental factors,
climate [22] as well as due to milling, storage, and manipulation. The proximate composition of
buckwheat, rice flours and oat flakes is similar to that reported by [23], being good sources of proteins,
lipids and minerals for human diet. Due to their nutritive value these flours are often selected as raw
materials for nutritionally balanced biscuits.
Molecules 2020, 25, 442 3 of 17

Table 1. Proximate composition of spent malt rootlets (MRF), whole wheat spelt flour (SWF), buckwheat
flour (BF), whole rice flour (WRF), oat flakes flour (OFF), and chia seeds flour (CF) used as raw materials
in enriched biscuits formulation.

Parameters MRF SWF BF WRF OFF CF

Moisture b 14.27 ± 0.3 e d 10.5 ± 0.4 c de 6.41 ± 0.1 a
8.2 ± 0.6 12.3 ± 0.2 13.3 ± 0.3
Proteins 35.5 ± 0.3 d 15 ± 0.3 bc 13.7± 0.2 b 8.5 ± 0.1 a 13.8 ± 0.2 b 15.9 ±0.2 c
Lipids 1.9 a ± 0.3 a 2.9 ± 0.3 a 2.8 ± 0.23 a 2.5 ± 0.3 a 6.1 ± 0.5 b 29.3 ± 0.2 c
Carbohydrates 46.70 ± 1.4 a 66.59 ± 1.0 bc 69.10 ± 0.21 bc 77.6 ± 0.6 c 63.70 ± 1.4 b 46.19 ± 0.1 a
Ash 7.7 ± 0.2 c 1.24 ± 0.2 ab 2.1 ± 0.3 ab 0.9 ± 0.1 a 3.1 ± 0.2 b 2.2 ± 0.2 ab
Each value was the mean of duplicate measurements; a–e Different superscripts in a row indicate significant
difference within samples (p < 0.05).

Regarding the MRF, it’s proximate composition was in the typical ranges as it was reported by
literature; the protein content of 35.5% and the total lipid amount (1.9%) are close to the values reported
by [7]. The carbohydrates content reached a similar value (46%) to that reported by [24], while the ash
content was similar with the amount reported by [25].
It has been reported that normal biscuits are nutritionally deficient as they are low in protein,
dietary fiber, as well as vitamins and minerals. Several studies have reported on improving the
nutritional quality of biscuits by using protein rich ingredients like oil seed meals [26], pulses [27],
and industrial by-products like wheat germ [28]. Recently, a few studies have reported the use of
multigrain to improve the nutritional quality of bread and other traditional products [29]. These
results could sustain the selection of these raw materials, including the spent malt rootlets as a better
choice for producing baked goods with enhanced nutritional value. Further papers will report on the
techno-functional properties of the enriched biscuits with MRF.

2.2. Fatty Acids Methyl Esters Content of Spent Malt Rootlets Flour (MRF) and Whole Wheat Spelt
Flour (SWF)
In the present study, by using GS-MS analysis, a total number of 19 fatty acid methyl esters
(FAMEs) were identified, while 16 of them where identified in MRF samples and only 13 in the SWF
samples, as reported in Table 2. Eight saturated fatty acids (SFAs) were found, with palmitic acid
being the predominant one in both analyzed flours but with an increased concentration for spent
malt rootlets flour. Regarding monounsaturated fatty acids (MUFAs), seven compounds were found,
among which, oleic acid was the major one. The content of oleic acid was identified as double in the
case of whole spelt flour, around of 24.66%, comparing to MRF. Concerning polyunsaturated fatty
acids (PUFAs), four compounds were found, standing out both linoleic and linolenic acids as major
compounds. Spent malt rootlets flour recorded high values of linoleic and linolenic acids (35.617%
and 32.64%, respectively). In the case of whole spelt flour only linoleic acid reached 26.696%, while
linolenic acid content was 4.426%. Similar results were reported by [7], showing that linoleic, linolenic,
palmitic and oleic are the main fatty acids from spent malt rootlets.
Regarding the lipid content of spelt wheat flour [30], reported that compared to the common
wheat, the lipid’s spelt content is higher and varies between 7–30% and palmitic acid, oleic acid, linoleic
acid and linolenic acid are the main fatty acids from spelt wheat.
By these results, MRF show a good fatty acids profile with high amount of PUFAs (ω−6 and
ω−3), essential fatty acids that must be derived from the diet, cannot be made by humans, and other
mammals mainly because of the lack of endogenous enzymes such as delta 12 and delta 15, accordingly
to [31] and [32].
Molecules 2020, 25, 442 4 of 17

Table 2. Comparative fatty acids methyl esters content (% of total fatty acids methyl esters) of spent malt rootlets flour (MRF) and whole wheat spelt flour (SWF).

Shorthand FA (% From Total FA) Samples

Fatty Acids Systematic Names Fatty Acids Trivial Name Type
Nomenclature SWF MRF
10:0 Decanoic acid Capric acid SFA 0.58 ± 0.21 0.31 ± 0.03
10:1(n-6) cis-4-Decenoic acid Obtusilic acid MUFA, ω−6 0.05 ± 0.01 0.14 ± 0.04
12:0 Dodecanoic acid Lauric acid SFA 1.13 ± 0.21 0.69 ± 0.17
15:0 Pentadecanoic acid Pentadecylic acid SFA 0.19 ± 0.15 0.42 ± 0.16
a15:0 Methyl-tetradecanoic acid Sarcinic acid SFA 0.15 ± 0.03 nd
16:0 Hexadecanoic acid Palmitic acid SFA 23.23 ± 0.11 30.50 ± 0.36
16:1(n-7) cis-9-Hexadecenoic acid Palmitoleic acid MUFA 0.69 ± 0.09 0.26 ± 0.08
17:0 Heptadecanoic acid Margaric acid SFA 3.24 ± 0.05 0.03 ± 0.01
18:0 Octadecanoic acid Stearic acid SFA 4.11 ± 0.02 1.45 ± 0.17
tr 18:1(n-9) trans-9-Octadecenoic acid Elaidic acid MUFA, ω-9 nd 0.09 ± 0.01
18:1(n-7) cis-11-Octadecenoic acid Vaccenic acid MUFA nd 1.15 ± 0.29
18:1(n-9) cis-9-Octadecenoic acid Oleic acid MUFA, ω-9 24.66 ± 0.04 12.13 ± 0.09
18:2(n-6) 9,12-Octadecadienoic acid Linoleic acid PUFA, ω−6 26.69 ± 0.03 35.61 ± 0.26
18:3(n-3) 9,12,15-Octadecatrienoic acid Linolenic acid PUFA, ω−3 4.42 ± 0.19 32.64 ± 0.38
20:0 Eicosanoic acid Arachidic acid SFA 0.11 ± 0.06 nd
20:4(n-6) 5,8,11,14-Eicosatetraenoic acid Arachidonic PUFA, ω−6 nd 0.79 ± 0.19
22:1(n-9) cis-13-Docosenoic acid Erucic acid MUFA, ω-9 nd 0.38 ± 0.15
22:6(n-3) 4,7,10,13,16,19-Docosahexaenoic acid Docosaheptaenoic acid PUFA, ω−3 nd 1.16 ± 0.53
P
SFAs - - - 32.63 ± 0.78 33.40 ± 0.90
P
MUFAs - - - 25.40 ± 0.14 14.15 ± 0.66
P
PUFAs - - - 31.11 ± 0.22 70.20 ± 1.36
P P
PUFAs/ MUFAs - - - 1.22 ± 1.57 4.96 ± 2.06
P P
PUFAs/ SFAs - - - 0.95 ± 0.28 2.10 ± 1.51
Note: Each value was the mean of triplicate measurements. FA-fatty acid, SFA-saturated fatty acids, PUFA-polyunsaturated, MUFA-monounsaturated fatty acids, nd-not detected; Each
value was the mean of duplicate measurements.
Molecules 2020, 25, 442 5 of 17

However, the presence of fatty acids in MRF contributes also to the generation of volatiles in
biscuits during the manufacturing process. Linoleic and linolenic acids are usually oxidized by
lipoxygenase action, resulting hydroperoxides which are unstable and are gradated during baking,
mainly into hexanal and hexenal. Lipoxygenase is more active in dough without yeast, more oxygen
remain disposable for this enzyme to transform lipids into aldehydes, ketones, and esters [33].
P
Out of the total of fatty acids, the high content of PUFAs in MRF is noticeable in a ratio of 4.95,
compared to 1.22 in the SWF, this proves the positive effect for health.

2.3. Volatile Compounds of Spent Malt Rootlets Flour (MRF) and Whole Wheat Spelt Flour (SWF)
In order to achieve this goal, a total number of 21 aroma compounds were analyzed by using
ITEX/GS-MS, as shown in Table 3, which were divided into five classes for an easier discussion:
alcohols, aldehydes, ketones, terpenes and terpenoids, others.

Table 3. Volatile compounds of spent malt rootlets flour (MRF) and whole wheat spelt flour (SWF) * .

RI (Retention
Volatile Compounds Perceived Flavour SWF (%) MRF (%)
Indices)
Alcohols
3-methylbutan-1-ol 736 Whiskey, malt, burnt - 40.21 ± 0.23
2-methylbutan-1-ol 739 Alcoholic, winey - 4.02 ± 0.49
pentan-1-ol 759 Fruity, sweet - 3.25 ± 0.16
Ethereal, Oil, alcohol,
hexan-1-ol 851 green, Fruity, Sweet, - 12.72 ± 0.38
Woody, Floral
oct-1-en-3-ol 975 Mushroom, herbal, earthy - 1.46 ± 0.23
Aldehydes
Fresh, Green, Fatty,
Hexanal 801 Aldehydic, Grass, Leafy, 35.96 ± 0.37 8.36 ± 0.51
Fruity, Sweaty
Fresh, Aldehydic, Fatty,
Heptanal 903 - 0.88 ± 0.49
Green, Burgundy, Grass
Aldehydic, rose, waxy,
Nonanal 1106 6.80 ± 0.28 -
citrus, orange, floral
2-methylpropanal - Wine, malt 49.20 ± 0.47 -
Ketone
Fruity spicy, sweet herbal,
heptan-2-one 895 - 2.09 ± 0.39
coconut woody
octan-3-one 965 Herbal - 2.94 ± 0.65
1-phenylethanone 1041 Almond, floral 8.02 ± 0.32 -
Ethanone - Whiskey - 0.29 ± 0.21
2-methylpentan-3-one - Mint - 0.25 ± 0.33
Terpenes and terpenoids
p-Cymene 1028 Citrus, sweet, herbal, spicy - 0.58 ± 0.66
Limonene 1031 Citrus, mint - 6.08 ± 0.48
Others
3-methylbutyl acetate 877 Sweaty, fruity, solvent - 0.54 ± 0.27
Nonane 900 Solvent - 0.68 ± 0.48
(E)-3,5,5-trimethylhex-2-ene 969 Solvent - 2.47 ± 0.77
2-pentylfuran 993 Buttery, green beans - 10.25 ± 0.11
(methyldisulfanyl)
- Sulfurous, onion, cabbage - 1.45 ± 0.24
methane
Each value was the mean of duplicate measurements.

Compared to whole wheat spelt flour, a greater number of volatile compounds were separated in
MRF samples. In MRF, a total number of 18 volatiles were identified, while SWF had only 4 identified
volatile compounds. The main MRF identified aroma compounds from the alcohols group were
3-methylbutan-1-ol (40.21%), 2-methylbutan-1-ol (4.02%), and pentan-1-ol (3.25%), while for SWF, no
compound from the alcohol class was identified.
Molecules 2020, 25, 442 6 of 17

The technological steps in processing barley into malt, such as steeping, germination and drying
of the germinated seeds involve several reactions which are responsible for the generation of an
enhanced aroma profile [34,35]. The germination temperature has a strong influence on the content
of volatile compounds. The high amount of 3-methylbutan-1-ol, 2-methylpentan-3-one is generally
correlated with germination temperature and green malt moisture content, being independent on the
time germination, according to [36].
Hexanal (8.36%) was the predominant compound of the aldehydes group. From the ketone
class in MRF, compounds like heptan-2-one, ethenone, 2-methylpentan-3-one and octan-3-one were
identified, while in WSF only 1-phenylethanone (8.02%) was identified out of the ketone group.
1-phenylethanone is characterized by [37] as having a sweet, floral and almond aroma. Limonene
(6.08%) and p-cymene were the volatile compounds from terpenes and terpenoids group, identified in
MRF sample. 2-pentylfuran (10.25%) was the only volatile compound found in MRF representing the
furans group, characterized by [38] as having an odor perception of green bean, butter. The aroma
profile of spent malt rootlets is showed in Figure 1, pointing out that alcohols contributed mainly by
giving whiskey, malt and alcoholic flavor notes as the main perceived aroma.
Molecules 2020, 25, x; doi: 6 of 16

Figure 1. Aroma profile of spent malt rootlets by main compounds classes identified during GC-MS
Figure 1. Aroma profile of spent malt rootlets by main compounds classes identified during
analysis.
GC-MS analysis.

Whole spelt flour

Whole spelt flourhad
hadasasaldehydes
aldehydes compounds
compounds 2-methylpropanal
2-methylpropanal (49.20%),
(49.20%), hexanal
hexanal (35.96%)
(35.96%) and
and
nonanenonane (6.80%),
(6.80%), probablyprobably
derivedderived by lipoxygenase
by lipoxygenase actionmilling
action during during andmilling and maturation
maturation processes.
processes.
2.4. Fatty Acids Methyl Esters Content of Final Enriched Biscuits
2.4. Fatty Acids Methyl Esters Content of Final Enriched Biscuits
In the enriched biscuits, a total of 12 fatty acids methyl esters (FAMEs) were identified, from
which Inseven
the enriched biscuits,
were classified asabeing
total saturated
of 12 fatty(SFAs),
acids three
methyl as esters
MUFAs, (FAMEs)
and twowere identified,
as PUFAs. from
The main
which
compoundssevenfrom
werethe classified
saturated as group
being were
saturated (SFAs),
myristic acid three as MUFAs,
reaching around of and
11%two as PUFAs.
of total The
fatty acids,
main compounds
followed by lauricfrom
acidthe
andsaturated group
capric acid. Fromwere myristic
PUFAs acid
acids, reaching
linoleic acidaround
reached of the
11%highest
of totalvalue
fatty
acids, followed
increasing by lauric
significantly acid
with theand
MRF capric acid.inFrom
addition PUFAs
biscuits. acids,
Also, linoleic
linolenic acidacid
andreached the reached
myristoleic highest
value
higherincreasing
contents assignificantly with the
the MRF content MRF addition
increased in biscuitsin biscuits. Also,As
formulation. linolenic
could be acid
seen and
bymyristoleic
comparing
reached
the fattyhigher contents(Tables
acids profiles as the 2MRF
and content increased
4), linoleic acid was in identified
biscuits formulation. As could
in a high amount be seen
in the by
control
comparing the fatty acids profiles (Tables 2 and 4), linoleic acid
sample too, highlighting that spelt wheat flour also contributed to this content. was identified in a high amount in
the control sample too, highlighting that spelt wheat flour also contributed to this content.
Fat is a key component in aroma perception being involved also in the texture of the final baked
goods and improving the sensory features such as the mouth-feel [39]. Lipid oxidation is involved in
the final characteristics of the food, such as aroma, taste, nutritional value, color, and texture [40].
According to [41], alcohols are formed mainly from the lipids through enzymatic reaction catalyzed
by lipoxidase. Also, by auto-oxidation of unsaturated fatty acids (C18:1, C18:2) short chain aldehyde
Molecules 2020, 25, 442 7 of 17

Table 4. Fatty acids methyl esters content (% of total fatty acids methyl esters) in enriched biscuits with spent malt rootlets in different concentrations.

Shorthand Fatty Acid’s

Type B0 B5 B10 B15 B20 B25
Nomenclature Name
4:0 Butyric SFA 1.75 ± 0.05 d 1.11± 0.11 a 1.19 ± 0.08 b 1.51 ± 0.05 c 1.77 ± 0.13 d 1.81 ± 0.09 d
6:0 Caproic SFA 0.90 ± 0.09 a 0.92 ± 0.17 a 0.98 ± 0.33 a 0.86 ± 0.06 a 0.98 ± 0.01 a 1.41 ± 0.09 b
8:0 Caprylic SFA 0.73 ± 0.23 b 0.83 ± 0.04 b 0.85 ± 0.23 b 0.84 ± 0.02 b 0.04 ± 0.18 a 0.04 ± 0.08 a
10:0 Capric SFA 2.30 ± 0.17 a 2.38 ± 0.02 a 2.48± 0.09 a 3.39 ± 0.24 b 2.31 ± 0.09 a 2.24 ± 0.05 a
10:1 Decenoic MUFA 0.24 ± 0.55 a 0.22 ± 0.19 a 0.22 ± 0.12 a 0.38 ± 0.01 b 0.25 ± 0.07 a 0.24 ± 0.81 a
12:0 Lauric SFA 3.56 ± 0.22 a 3.34 ± 0.27 a 3.37± 0.04 a 3.75 ± 0.06 a 4.63 ± 0.04 b 5.10 ± 0.04 c
14:0 Myristic SFA 11.06 ± 0.08 c 11.05 ± 0.15 c 11.04 ± 0.02 c 11.04 ± 0.17 c 11.02 ± 0.12 b 10.95 ± 0.01 a
14:1 Myristoleic MUFA 1.19 ± 0.03 ab 1.11 ± 0.11 a 1.18 ± 0.23 ab 1.24 ± 0.08 bc 1.33 ± 0.22 c 1.84 ± 0.21 d
16:0 Palmitic SFA 0.44 ± 0.02 c 0.35 ± 0.05 a 0.41 ± 0.11 b 0.41 ± 0.23 b 0.40 ± 0.03 b 0.40 ± 0.25 b
18:3(n-3) Linolenic PUFA ω-3 1.98 ± 0.20 a 1.92 ± 0.05 a 1.99 ± 0.08 a 2.38 ± 0.06 a 2.11 ± 0.07 a 4.60 ± 0.04 b
18:1(n-9) Oleic MUFA, ω-9 1.26 ± 0.22 c 1.01 ± 0.14 a 1.18 ± 0.02 b 1.17 ± 0.39 b 1.15 ± 0.03 b 1.18 ± 0.44 b
18:2(n-6) Linoleic PUFA, ω−6 29.13 ± 0.04 a 32.36 ± 0.03 b 32.62 ± 0.01 b 38.78 ± 0.06 c 47.13 ± 0.03 d 51.67 ± 0.07 e
20.74 ± 0.86 bc 19.98 ± 0.81 a 20.32 ± 0.90 ab 21.80 ± 0.83 d 21.15 ± 0.60c 21.95 ± 0.61d
P
SFAs - -
2.69 ± 0.80 bc 2.34 ± 0.74 a 2.58 ± 0.09 b 2.79 ± 0.48 c 2.73 ± 0.32 bc 3.26 ± 1.46 d
P
MUFAs - -
31.11 ± 0,24 a 34.28 ± 0.28 b 34.61 ± 0.09 b 41.16 ± 0.12 c 49.24 ±0.10 d 56.27 ± 0.11 e
P
PUFAs - -
11.57 ± 0.30 a 14.65 ± 0.38 c 13.41 ± 0.35 b 14.75 ± 0.25 c 18.04 ± 0.31 d 17.26 ± 0.08 d
P P
PUFAs/ MUFAs - -
1.5 ± 0.28 a 1.72 ± 0.10 b 1.70 ± 0.10 b 1.89 ± 0.14 c 2.33 ± 0.17 d 2.56 ± 0.18 e
P P
PUFAs/ SFAs
0.13 ± 0.93 b 0.12 ± 0.54 a 0.13 ± 0.29 b 0.13 ± 0.58 b 0.13 ± 0.53 b 0.15 ± 2.39 c
P P
MUFAs/ SFAs
P+
P
P PUFAs 1.63 ± 1.21 a 1.83 ± 0.64 b 1.83 ± 0.39 b 2.02 ± 0.72 c 2.46 ± 0.70 d 2.71 ± 2.57 e
MUFAs/ SFAs
Note: FA-fatty acid, SFA-saturated fatty acids, PUFA-polyunsaturated, MUFA-monounsaturated fatty acids, biscuits samples with different amount of spent malt rootlets
(0%,5%,10%,15%,20%,25%). a–e Different superscripts in a row indicate significant difference within samples (p < 0.05). Each value was the mean of duplicate measurements.
Molecules 2020, 25, 442
x; doi: 8 of 16
17

Fat is aCompounds
2.5. Volatile key component in aroma
Content perception
in Enriched Biscuitsbeing involved also in the texture of the final baked
goods and improving the sensory features such as the mouth-feel [39]. Lipid oxidation is involved
A total number of 18 volatile compounds were identified in the enriched biscuits, by means of
in the final characteristics of the food, such as aroma, taste, nutritional value, color, and texture [40].
ITEX/GC-MS technique as shown in Table 5. Also, the perceived aroma was correlated with the
According to [41], alcohols are formed mainly from the lipids through enzymatic reaction catalyzed
literature data, the perceived flavor of each identified compound is showed in the same Table 5
by lipoxidase. Also, by auto-oxidation of unsaturated fatty acids (C18:1, C18:2) short chain aldehyde
[37,43,44].
(C6-C9) could be formed [18]. Hexanal is a typical compound of linoleic and arachidonic acids
Limonene, β-pinene, hexanal, 3-methylbutan-1-ol were the principal volatile compounds found
oxidation. It is often used as a marker of lipid oxidation.
in the final baked biscuits, influencing
P the taste by having a citrus, mint flavor together with fresh,
P
The ratio of fatty acids PUFAs/ MUFAs on MRF/SWF, Table 2, are correlated with enrich
green, fruity, sweet or pine, resine and turpentine
P flavours
P (Figure 2). Limonene has been reported
biscuits, as can be seen in Table 4, the ratio PUFAs/ MUFAs on B0 is 11.532, compared to 17.24 in
having chemo- preventive activity against different types of cancer such as breast or colorectal ones
B25, this ratio increases with the addition of MRF. According to [42], it seems that the PUFA/SFA ratio
at a concentration between of 0.5–12 g/m2/day [45]. The amount of limonene could be correlated with
alone is not suitable to predict the changes of plasma lipids level and that the PUFA + MUFA/SFA
the carotenoids amount, according to [41]. The high amount of limonene could be due to the high
ratio is a more suitable parameter for this purpose. Moreover, the authors [42] suggest that the main
content in carotenoids of chia seeds used in biscuits formulation, since it is present even in the
prerequisites for keeping low plasma and liver cholesterol are: (i) low MUFA/SFA ratio, (ii) high
control sample in relative high amount 24.76% and the limonene content increased progressively
PUFA/MUFA ratio, and (iii) PUFA + MUFA/SFA ratio not to exceed 2. From this point of view, the
with the MRF addition.
sample B0, B5, B10 and B15 are meeting the above-mentioned criteria, as can be seen in Table 4.
From the ketone group, 1-phenylethanone was the main volatile compound identified, being
characterized by havingContent
2.5. Volatile Compounds an almond, floral Biscuits
in Enriched flavor.
Hexanal, according to [38], is the end product of the lipoxygenases and hydroperoxide
A total number of 18 volatile compounds were identified in the enriched biscuits, by means of
isomerases which are involved in the oxidation of unsaturated fatty acids. Also [46], mentioned that
ITEX/GC-MS technique as shown in Table 5. Also, the perceived aroma was correlated with the literature
hexanal is the main product of linoleic acid autoxidation. This idea is supported by other authors
data, the perceived flavor of each identified compound is showed in the same Table 5 [37,43,44].
[41] who mentioned that unsaturated fatty acids could influence the amount of aldehydes and
Limonene, β-pinene, hexanal, 3-methylbutan-1-ol were the principal volatile compounds found
ketones in flours
in the final baked biscuits, influencing the taste by having a citrus, mint flavor together with fresh,
β-pinene with pine, turpentine and traces of mint, camphor and eucalyptus aromas and α-
green, fruity, sweet or pine, resine and turpentine flavours (Figure 2). Limonene has been reported
pinene with a citrus and spicy, woody pine aroma contribute significantly to the aroma profile of
having chemo- preventive activity against different types of cancer such as breast or colorectal ones at
enriched biscuits (Figure 2). α and β-pinene are important compounds used in food preservation,
a concentration between of 0.5–12 g/m2 /day [45]. The amount of limonene could be correlated with
due to their antimicrobial activity. More than that, α-pinene has been reported to have antitumor
the carotenoids amount, according to [41]. The high amount of limonene could be due to the high
activity on a concentration of 8 mg/L [47].
content in carotenoids of chia seeds used in biscuits formulation, since it is present even in the control
The presence of 3-methylbutan-1-ol in enriched biscuits could be explained by the spent malt
sample in relative high amount 24.76% and the limonene content increased progressively with the
rootlets addition since this compound was not identified in the control sample and MRF contained
MRF addition.
high amounts of this compound (Table 3).

Figure 2. Aroma profile of enriched biscuits with spent malt rootlets in different concentrations
Figure 2. Aroma profile of enriched biscuits with spent malt rootlets in different concentrations (0–25%).
(0–25%).

.
Molecules 2020, 25, 442 9 of 17

Table 5. Volatile compounds content of the enriched biscuits with spent malt rootlets.

RI
Volatile Compounds (Retention Perceived Flavour B0 B5 B10 B15 B20 B25
Indices)
Alcohols
3-methylbutan-1-ol 736 Malty, alcoholic whiskey - 1.34 ± 0.03 a 4.12 ± 0.09 b 7.45 ± 0.71c 8.41± 0.02 d 11.27 ± 0.01 e
Aldehyde
Fresh, Green, Fatty,
Hexanal 801 Aldehydic, Grass, Leafy, 4.51 ± 0.06 a 6.21 ± 0.25 b 9.29 ± 0.28 c 12.58 ± 0.29 d 25.06 ± 0.11 e 28.22 ± 0.26 f
Fruity, Sweaty
Almond, String, Sharp,
Benzaldehyde 960 07.1 ± 0.02 abc 0.39 ± 0.55 a 1.37 ± 0.07 bc 1.47 ± 0.01 c 0.6 ± 0.03 ab 3.9 ± 0.36 d
Sweet, Bitter, Cherry
Ketone
Fruity spicy, sweet herbal,
heptan-2-one 895 0.88 ± 0.02 b - - - 0.51± 0.33 a -
coconut woody
b bc cd b a
1-phenylethanone 1042 Almond, floral 1.34 ± 0.07 1.78 ± 0.02 2.49 ± 0.28 1.3 ± 0.56 0.39 ± 0.44 2.79 ± 0.08 d
Terpenes and terpenoids
α-Thujene 938 Green, herbal, woody - - - - 0.38 ± 0.55 -
Fresh, Sweet, Green, Woody,
α-Pinene 939 1.53 ± 0.05 a 2.77 ± 0.24 e
2.38 ± 0.45 d 2.27 ± 0.22 c
1.75 ± 0.22 b -
Earthy
β-Pinene 982 pine, resin, turpentine 21.81 ± 0.66 a 21.16 ± 0.21 a 27.76 ± 0.77 b 29.9 ± 0.12 c 36.78 ± 0.09 d 40.24 ± 0.03 e
Tropical, fruity with mango
β-Myrcene 992 - - - - 1.84 ± 0.21 -
shades, grassy
α-Phellandrene 1006 Citrus, woody, grassy, pepper - - - 1.19 ± 0.06 a
2.15 ± 0.44 b -
p-Cymene 1028 Citrus, sweet, herbal, spicy 2.98 ± 0.31 a 3.01 ± 0.23 a 4.17 ± 0.44 b 4.53 ± 0.03 b 4.68 ± 0.04 b 2.54 ± 0.22 a
Limonene 1031 Citrus, mint 24.76 ± 0.38 a 33.41 ± 0.56 b 35.22 ± 0.09 b 41.49 ± 0.37 c 55.91 ± 0.03 d 60.15 ± 0.45 e
Citrus, tropical, fruity, oily,
γ-Terpinene 1074 - - - - 3.02 ±0.05 -
woody
1,3,8-p-Menthatriene 1110 Woody, citrus, grassy - 4.39 ± 0.05 - - - -
Acids
Benzoic acid 1277 balsamic 1.14 - - - - -
Other compounds
2-pentylfuran 993 Butter, green beans - - - - - 3.32 ± 0.33
n.i. - 1.54 ± 0.22 a 2.01 ± 0.11 b 1.41± 0.46 a 1.98 ± 0.55 b - 1.79 ± 0.41 b
n.i. - 2.09 ± 0.44 bc 2.33 ± 0.51 c 2.13 ± 0.33 c 1.64 ± 0.38 b 0.81 ± 0.88 a 2.16 ± 0.22 c
Note: a–d different superscripts in a row indicate significant difference within samples (p < 0.05). Each value was the mean of duplicate measurements.
Molecules 2020, 25, 442 10 of 17

From the ketone group, 1-phenylethanone was the main volatile compound identified, being
characterized by having an almond, floral flavor.
Hexanal, according to [38], is the end product of the lipoxygenases and hydroperoxide isomerases
which are involved in the oxidation of unsaturated fatty acids. Also [46], mentioned that hexanal
is the main product of linoleic acid autoxidation. This idea is supported by other authors [41] who
mentioned that unsaturated fatty acids could influence the amount of aldehydes and ketones in flours.
β-pinene with pine, turpentine and traces of mint, camphor and eucalyptus aromas and α-pinene
with a citrus and spicy, woody pine aroma contribute significantly to the aroma profile of enriched
biscuits (Figure 2). α and β-pinene are important compounds used in food preservation, due to their
antimicrobial activity. More than that, α-pinene has been reported to have antitumor activity on a
concentration of 8 mg/L [47].
The presence of 3-methylbutan-1-ol in enriched biscuits could be explained by the spent malt
rootlets addition since this compound was not identified in the control sample and MRF contained
high amounts of this compound (Table 3).
Undoubtedly, the other ingredients from the biscuits formulation (multigrain mix, butter, malt
extract) also contributed to the final volatile derivatives content and to the biscuits aroma profile through
lipid oxidation, Maillard reactions, including Strecker degradation of the carbonyl compounds and
caramelization [33]. These ingredients act as a possible source of precursors for volatile compounds due
to their chemical composition of lipids and antioxidants, proteins and sugars [48]. The transformations
occurred mainly during baking but also during the resting period of the biscuit dough.

2.6. Pearson Correlation

In order to study the correlation between the % of spent malt rootlets addition and the volatile
compounds/fatty acids of the final baked products, Pearson correlation was run. In the final baked
goods, hexanal amount increased as the % of MRF increased having a total value of 28.22% in the
biscuits enriched with 25% MRF. The amount of hexanal in MRF (8.36%), increased 3.37 times in the
final baked goods with 25% MRF, having a Pearson correlation of 0.9558. The Pearson correlation
between the amount of 3-methylbutan-1-ol from the SMR and the final baked goods is 0.991 which
indicated a strong relationship between this raw material and the final products. Also, Limonene
content of the MRF influenced the amount of the final baked goods, showing a strong relation by
having a positive Pearson correlation of 0.9753. In contrast, from the fatty acids, only linoleic acid
reached a Pearson correlation value of 0.8266, followed by myristoleic (0.6593), linolenic (0.5425), lauric
and oleic acids with correlation values of 0.518 and 0.2435, respectively.

2.7. Sensory Analysis

The enriched biscuits showed significant differences for sensory descriptors compared to the
control sample. The enriched biscuits were characterized by more intense color, flavor, taste, odor
as the % of MRF increased. The biscuits with 15% MRF reached the highest hedonic score, having a
mean sensory score for the visual appearance (color), taste and flavor of 8.1, 8.2 and 8.3 respectively, as
showed in the Figure 3. Also, regarding the texture and overall acceptability, the panelists preferred
the biscuits with 15% MRF, having a total score of 7.9 and 8.1 respectively. This was also confirmed
by [13], who reported that an amount of 20% MRF in the biscuits caused the rejection of the product,
due to the flavor, appearance and texture.
the % of MRF increased. The biscuits with 15% MRF reached the highest hedonic score, having a
mean sensory score for the visual appearance (color), taste and flavor of 8.1, 8.2 and 8.3 respectively,
as showed in the Figure 3. Also, regarding the texture and overall acceptability, the panelists
preferred the biscuits with 15% MRF, having a total score of 7.9 and 8.1 respectively. This was also
Molecules 2020,by
confirmed 25,[13],
442 who reported that an amount of 20% MRF in the biscuits caused the rejection
11 of of
17

the product, due to the flavor, appearance and texture.

Figure 3. Hedonic scores for enriched biscuits with spent malt rootlets in different concentrations
Figure 3. Hedonic scores for enriched biscuits with spent malt rootlets in different concentrations
(0–25%).
(0–25%).

In the case
In the case of
of the
the lower
lower color
color hedonic
hedonic scores
scores reached
reached at
at 20%
20% and
and 25%
25% MRF
MRF addition
addition (6.5,
(6.5, 6.0,
6.0,
respectively),
respectively), the panelists described the biscuits as having a dark color. As the percentage of
the panelists described the biscuits as having a dark color. As the percentage MRF
of MRF
increased, the color
increased, the colorofofthe
thebiscuits
biscuitsdarkened.
darkened.This
This
is is
in in agreement
agreement with
with [6] [6]
whowho reported
reported that that
MRFMRFhad
an impact on the crust color of the bread. Also, the Maillard and caramelization reactions influenced
the final color of the biscuits, according to [49].
Regarding the biscuits taste and odor, panelists described an intense note of whisky, alcohol
partly imputable to the increment of 3-methylbutan-1-ol with the MRF addition. Also, for the samples
with more than 15% MRF, panelists indicated an unpleasant aldehydic taste imputable to the hexanal
amount which was negative correlated to the bread aroma by consumers [33]. Citrus, pine, and mint
notes were indicated by panelists as pleasant aromas for enriched biscuits and are positively correlated
with limonene and β-pinene.

3. Materials and Methods

3.1. Procurement of Raw Materials

Whole rice flour, buckwheat flour, whole wheat flour, chia seeds flour and oat flakes flour were
purchased from specialized stores in Cluj-Napoca, România. According to the producers, all the
ingredients meet the highest quality standards. Malt extract and natural lemon aroma were purchased
from SC. Ireks Pan SRL Bucures, ti, România. Barley dried malt rootlets were received from SC. Soufflet
Agro SRL Buzau, România and were obtained from the autumn barley Sebastian Ukraina variety in
2016. Barley malt rootlets are the dried shoots and rootlets of sprouted grain in the brewing process
and is considered a by-product [6,10,50]. All reagents were of analytical grade. Analytical reagents
and chemicals were purchased from Sigma Aldrich (Louis, MO, USA).

3.2. Malt Rootlets Flour Preparation

The dried rootlets were passed through a mill feeder processed by grinding to fine flour (<300
µm) on a Grindomix (Model GM200, Haan, Germany) laboratory mill at 10,000 rot/min for 50 s, fitted
with a 0.8 mm sieve and then homogenised by mixing. Barley malt rootlets flour (MRF) was subjected
by supplemented in different proportion to prepare five different digestive biscuits.
Molecules 2020, 25, 442 12 of 17

3.3. Proximate Composition Analysis of the Flours and Biscuits

The chemical characteristics were carried out according to AACC Approved Methods [51] like
moisture (44–15.02), lipids (30–25.01), ash (08–01.01), crude fibre (32–07.01) and protein were measured
using the Kjeldahl method (46–11.02), nitrogen to protein conversion factor was 5.7. Total carbohydrate
(%) content was calculated as the difference: 100 − (moisture + ash + proteins + lipids + crude fibres)
method reported in a previous work [52], and also reported by [53].

3.4. Preparation of Dough and Baking Biscuits

The formulation of biscuit dough was summarized in Table 6. The multigrain mix consisted of
whole meal rice flour, buckwheat flour, chia seeds flour and oat flakes flour in a ratio of: 40:30:20:10
(WRF:BF:CSF:OFF). The formulation used for biscuits manufacturing (Table 6) was calculated taking
into consideration a basis formed by spelt whole wheat flour and a multigrain mix in the following
ratios 40:60 (SWF:MG). Malt rootlets flour (MRF) at 5%, 10%, 15%, 20% and 25% supplementation
levels, were incorporated into multigrain mix to produce composite flour, according to Table 6.

Table 6. Formulation used for the preparation of enriched biscuits.

Components (g) B0 (Control) B5 B10 B15 B20 B25

Spelt whole wheat flour (SWF) 40 40 40 40 40 40
Multigrain mix (MG) 60 55 50 45 40 35
Malt rootlets flour (MRF) 0 5 10 15 20 25
Butter 40 40 40 40 40 40
Malt extract 27 27 27 27 27 27
Baking powder 0.5 0.5 0.5 0.5 0.5 0.5
Lemon juice 1 1 1 1 1 1
Natural lemon aroma 0.5 0.5 0.5 0.5 0.5 0.5
Salt 0.2 0.2 0.2 0.2 0.2 0.2

To obtain the control samples (B0), all dry ingredients (whole meal rice flour (WRF), buckwheat
flour (BF), spelt whole wheat flour (SWF), chia seeds flour (CSF) and oat flakes flour (OFF)) were mixed
together in a mixer (KitchenAid® Precise Heat Mixing Bowl., Ohio, OH, USA), butter and malt extract
were previously emulsified and then added by incorporation into the flour mix and then combined
with the rest of the ingredients previously emulsified as well (baking powder, lemon juice, natural
lemon aroma and salt) according to the recipe presented. The dough was mixed for 10 min to obtain a
homogeneous consistency and then placed into the rest room at 6 ◦ C, over a period of 30 min. The
dough was then rolled out and 5 mm thick biscuits with a 60 mm diameter were formed and baked at
200 ◦ C for 12 min. The biscuits enriched with the malt rootlets flour were codified as follows: B5, B10,
B15, B20 and B25, according the amount of MRF used for supplementation. These were then stored in
glass containers for two months, protected from light, and at a room temperature of 21 ◦ C. The results
of all the experiments are given as the average of replicates. Each replicate (biscuit) was obtained from
separately prepared batches of dough. The analyses were carried out on fresh biscuits (on the day of
baking). The procedure of its preparation was adapted from another similar study [53].

3.5. Determination of Fatty Acid Composition

3.5.1. Total Lipid Determination

Lipids were extracted in petroleum ether using a Soxtest Raypa SX-6 MP (Barcelona, Spain)
apparatus. Three g of crushed biscuits sample was introduced in cartridges and 50 mL petroleum
was used for each sample extraction. Parameters were set as follows: temperature −75 ◦ C, time of
extraction −50 min. Samples were dried to constant weight. Total fat was expressed as percentage
from the sample (% w/w).
Molecules 2020, 25, 442 13 of 17

3.5.2. Fatty Acids Profile by GC-MS Analysis

Fatty acid methyl esters (FAMEs) from total lipids were analyzed using a Shimadzu
GCMS-QP2010PLUS apparatus and an AT-WAX column (30 m, 0.32 mm i.d., 1 µm thickness),
(Shimadzu Scientific Instruments, Kyoto, Japan) belonging to the Interdisciplinary Research Platform
of Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania.
The solvent used for FAMEs was hexane, while the carrier gas (helium) was used at a flow rate of
1.00 mL/min and a linear velocity of 37.8 cm/s. Derivatization was performed for 1 h at 80 ◦ C in
an ultrasonic bath. An aliquot of 0.1 g sample was treated with 3 mL boron trifluoride methanol
solution 20% (w/v). After cooling, 2.5 mL NaCl solution 10% (w/v) was added and methyl esters were
extracted in 2 mL hexane, the organic layer being separated by centrifugation at 1006 G for 15 min
using centrifuge Z36 HK (Hermle Labortechnik GmbH, Wehinge, Germany) for 15 min. The hexane
solution (1 µL) was injected in apparatus. Samples were analyzed keeping the column initially at 140
◦ C for 10 min and then increasing the temperature with 7 ◦ C/min up to 250 ◦ C and maintaining at this

temperature for 10 min (total run: 35.71 min). Split ratio was 1:10 and injection port temperature was
set at 250 ◦ C. The ion source and interface temperatures were 210 ◦ C and 255 ◦ C.
FAME peaks were identified using NIST05 library and quantified by area normalization method.
Some compounds were unidentifiable due to the lack of authentic samples and library spectra of the
corresponding molecules. The percentage of various lipidic compounds was determined by reporting
the peak area corresponding to a specific compound to the total peak area (for all identified constituents)
of chromatograms [54].

3.6. Extraction and Analysis of Volatile Compound

The analysis of volatile compounds was carried out on a GCMS QP-2010 (Shimadzu Scientific
Instruments, Kyoto, Japan) gas chromatograph-mass spectrometer equipped with a CombiPAL
AOC-5000 auto-sampler (Kyoto, Japan). The extraction of volatile compounds from samples was
performed using the in-tube extraction technique (ITEX) [43] with some modification. Three grams of
crushed biscuit samples were introduced into 20 mL headspace vial, incubated at 60 ◦ C for 20 min
under continuous stirring; the volatile compounds in the gas phase were repeatedly adsorbed (30
extractions) by fibre syringe ((ITEX-2TRAPTXTA, Tenax TA 80/100 mesh) and then were thermally
desorbed directly into the GC-MS injector.
The volatile compounds were separated on a Zebron ZB-5MS (Phenomenex) capillary column of
(30 m, 0.32 mm i.d., 1 µm thickness). The carrier gas was helium, the flow rate was 1 mL/min and the
split ratio 1:5. The chromatographic column temperature program has been set by: 35 ◦ C (held for 5
min) rising to 155 ◦ C with 7 ◦ C/min and then heated to 260 ◦ C with 10 ◦ C/min and held for 5 min. The
injector, ion-source and interface temperatures were set at 250 ◦ C. The MS mode was electron impact
(EI) at ionization energy of 70 eV. The mass range scanned was 40–400 m/z. The volatile compounds
were tentatively identified based on the spectra of reference compounds from NIST27 and NIST147
mass spectra libraries (with a similarity of minimum 85%) and verified by comparison with retention
indices drawn from [55] or [56]. All peaks found in at least two of the three total ion chromatograms
(TIC) were taken into account when calculating the total area of peaks (100%) and the relative areas of
the volatile compounds [23,38].

3.7. Sensory Evaluation

The sensory characteristics of the biscuit sample was carried out by 60 panellists, 30% male and
70% female, range: 20–65 years. Each panellist analysed two biscuits from each sample, taking into
account the texture, flavour, taste, overall acceptability and appearance on a nine-point Hedonic scale,
in the following sequence: 1–4 represent negative sensations, 5 was neither like nor dislike, and 6–9,
positive sensations, 9 meaning extremely like.
Molecules 2020, 25, 442 14 of 17

3.8. Statistical Analysis

The results of three independent (n = 3) assays performed with replicates each were expressed
as means ± standard deviations. Data were compared using Duncan multiple comparison test by
using SPSS version 19 software (p ≤ 0.005). Pearson correlation was also done to correlate the MRF
percentage of addition with the volatile compounds content.

4. Conclusions
To the best of our knowledge, there are very few works reporting on the relationship between the
spent malt rootlets addition in baked goods and their fatty acids, volatile and sensory profiles. Spent
malt rootlets are by-products of the brewing industry and are a good source of valuable nutrients.
However, their specific aroma profile could limit their addition in baked products. The results of this
study reveal the fatty acids and volatile profile of spent malt rootlets. Moreover, the same profiles are
determined for enriched biscuits with spent malt rootlets pointing out the contribution of fatty acids to
the generation of aroma compounds. Also, a correlation between the addition of spent malt rootlets and
the aroma and sensory profile of biscuits was done in order to obtain information about how different
volatile compounds influence the consumer’s preferences and finally, what is the optimum percentage
of addition. Future studies will report on the nutritional and quality parameters of multigrain biscuit
enriched with spent malt rootlets.

Author Contributions: Conceptualization, M.S.C. and A.P. (Adriana Păucean); methodology, A.P. (Anamaria
Pop), S.A.S., E.A., S.M.M., and M.B.; software, M.S.C.; validation, S.A.S., and E.A. formal analysis, A.P. (Adriana
Păucean), S.M.M. and S.A.S.; writing—original draft preparation, M.S.C. and A.P. (Anamaria Pop); writing—review
and editing, A.P. (Anamaria Pop) supervision, S.M.; project administration, M.S.C.; funding acquisition, M.S.C.
All authors have read and agree to the published version of the manuscript.
Funding: This research was funded by Executive Unit for Financing Higher Education, Research, Development
and Innovation (UEFISCDI) within the framework of PNCDI III-PN-III-P2-2.1-CI-2018-1503, grant No: 260/CI/2018
and the publication was supported by funds from the National Research Development Projects to finance excellence
(PFE)-37/2018–2020 granted by the Romanian Ministry of Research and Innovation.
Acknowledgments: The authors acknowledge the technical support provided by doctoral research project of the
researcher Monica Bota.
Conflicts of Interest: The authors have no conflict of interest regarding the content of this paper.

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