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Article

Analysis of Volatile Compounds and Sugar Content in Three Polish Regional Ciders with Pear Addition

1
Department of Human Nutrition and Diet Therapy, Branch Faculty of University of Zielona Góra in Sulechów, ul. Armii Krajowej 51, 66-100 Sulechów, Poland
2
Department of Fermentation and Cereals Technology, Faculty of Biotechnology and Food Science, Wrocław University of Environmental and Life Science, Chełmońskiego 37 Street, 51-630 Wrocław, Poland
3
Department of Chemistry, Faculty of Biotechnology and Food Science, Wrocław University of Environmental and Life Sciences, C.K. Norwida Street 25, 50-375 Wrocław, Poland
*
Author to whom correspondence should be addressed.
Molecules 2020, 25(16), 3564; https://doi.org/10.3390/molecules25163564
Submission received: 22 July 2020 / Revised: 2 August 2020 / Accepted: 4 August 2020 / Published: 5 August 2020

Abstract

:
Aroma plays important part in cider acceptability to the consumer. In this study, techniques such as headspace solid-phase microextraction (HS-SPME), which has been coupled with gas chromatography and mass spectrometry, have been used to assess what changes in the volatilome occur during fermentation of three apple cultivars (Cortland, Gala, Idared) with and without addition of pear (Konferencja) juice addition. Analysis of volatiles has shown that temperature of fermentation, apple variety and pear juice addition have significant influences on the volatile compositions of the acquired ciders. Ciders prepared in laboratory conditions fermented at 15 °C were characterized by a greater share of esters, such as ethyl hexanoate, ethyl decanoate and ethyl dodecanoate, in volatile profile (66.24–79.58%) than ciders fermented at 20 °C (58.81–77.22%). Ciders fermented at a higher temperature were characterized by a greater share of alcohols, such as phenylethyl alcohol and hexan-1-ol (18.34–36.7%) than ciders fermented at a lower temperature (16.07–25.35%). In the ciders prepared from pear (20% w/w) and apple (80% w/w) juice, the presence of esters, such as ethyl (2E, 4Z)-deca-2,4-dienoate, characterized by a pear aroma, could be noted.

1. Introduction

Cider (also called “hard cider,” in North America) is an alcoholic drink obtained by alcoholic fermentation of apple juice, although many ciders nowadays are produced from diluted apple juice concentrate. Cider can be consumed as a final product, albeit it can be also used as a substrate for the production of cider vinegar, produced by acetic acid bacteria or by the means of distillation, for the production of apple brandies, such as Calvados in Normandy and Lambig in Brittany. Cider is currently produced in many countries on the North American, Asian, European and Australian continents, which is why its individual types may differ significantly, depending on the region or country wherein they are produced [1]. In Poland, ciders are made by alcoholic fermentation of an apple juice or diluted apple juice concentrate with the possibility of adding pear juice in an amount of up to 40% by volume [2]. Apples and pears are the sources of many phytochemicals and vitamins, which are beneficial for human health, but health-promoting properties of the cider are not the most important ingredients which have impacts on the aroma of the cider [3,4,5,6,7]. Volatile compounds play a crucial part in creating the fragrance of the cider and other alcoholic beverages [8]. There are many factors which influence these characteristics, such as the variety of apple used, the degree of fruit ripeness, the conditions under which apple trees are grown, the processes used to extract apple juice and the fermentation temperature and species of microorganisms used for fermentation [9]. The main chemical groups of volatile compounds identified in cider are alcohols, acids and esters, such as 1-butanol, 1-hexanol, 3-methylbutyl acetate, 2-phenylethyl acetate and butyl acetate naturally occurring in fruits and produced by microorganisms that are used to ferment cider [10]. Volatile compounds derived from ciders have been studied by many scientists, but there still exists a gap in knowledge about the impact of the addition of pears or pear juice on the aroma of cider [11]. These studies are important, among others, because Polish legislation does not allow for pear fragrance to dominate apple aroma in ciders with the addition of pear juice [2]. The aim of the study was to investigate the composition of volatile compounds in ciders produced from three varieties of regional apples and their mixtures with pear juice during fermentation at two different temperatures.

2. Results

2.1. Volatile Compounds in Apple and Apple/Pear Juices

In this study, volatile compositions of apple juice and apple juice and pear juice mixes from three apple cultivars and one pear cultivar were assessed by means of gas chromatography coupled with mass spectrometry. In the apple juices, 43 volatile compounds were identified (Table 1) and in the pear juice/apple juice mixes, 46 volatile components could be recognized. They were divided into five chemical groups. The largest number of identified compounds in apple juices were esters (27 compounds) and the second largest group were alcohols (10 compounds). Other chemical groups consisted of aldehydes (three compounds), sesquiterpenes (two compounds) and acids (and compound). Pear and apple juice mixes also contained five different chemical groups of components, but 30 esters instead of 27 were identified. Idared and Konferencja juices mixture (IK) had the highest relative abundance of esters out of the analyzed juices (84.37% of peaks area). The addition of pear increased contributions of esters in the aroma profile of this juice, because esters constituted 80.92% of peak area of Idared juice (I). The lowest relative concentration of esters, which equaled 28.63%, was identified in the Cortland and Konferencja juice mixture (CK) and was similar to the relative concentration of esters in Cortland juice (C)—29.94%. In Gala variety juice (G) and Gala and Konferencja juices (GK) mixture, esters constituted, respectively, 65.54% and 66.43%. Second largest chemical group of compounds which were identified in juices was alcohols. Alcohols constituted the greatest share of volatiles in C (60.57%) and CK (65.17%), over two times greater than esters in these juices. The lowest abundance of alcohols was found in IK (9.71%) and the next in GK (13.38%). The same apple juices, without the contribution of pear juice, were characterized by greater amounts of alcohols: 14.35% in I and 23.22% in G. A greatest amount of aldehydes was found out in GK (10.35%), and in the rest of the samples aldehydes constituted only a minuscule amount of volatile components (0.28–1.37%). Addition of pear juice to each apple juice resulted in a lower relative share of sesquiterpenes in the composition of volatiles. Sesquiterpenes constituted the largest part of volatile composition in C (4.37%) and CK (3.28%) compared to the other samples, wherein the contribution of sesquiterpenes to aroma profile was lower, from 1.96% in I to 1.07% in GK. Acids, with only one compound in this group, were an important part of the aromas of G and GK, wherein they constituted, respectively, 9.28% and 8.59% of total volatiles. In other juices, the amount of acids was 2–4 times lower than in the juices with higher concentrations (2.41–4.3%). For the I and IK, the main aroma-forming compounds were ethyl 2-methylbutanoate, 2-methylpropyl acetate and hexyl acetate, which are associated with sweet, appley and banana aromas [12]. G and GK were also characterized by large concentrations of hexyl acetate in the overall aromatic profile. Main components of the C and CK were not only esters, such as 2-methyl propyl acetate acetate and butyl hexanoate, but alcohols such as (Z)-hex-2-en-1-ol and (E)-hex-2-en-1-ol. Three compounds, characterized by pear aroma, were not present in the samples without pear addition, but were identified in all samples with the addition of pear juice [13]. These were three esters: ethyl (Z)-dece-4-noate, methyl (2E,4Z)-deca-2,4-dienoate, ethyl (2E,4Z)-deca-2,4-dienoate.

2.2. Volatile Compounds in Ciders

Gas chromatography and mass spectrometry allowed for the identification of 32 volatile compounds in ciders (Table 2). As in the case of juices, volatile components identified in ciders could be divided into five different chemical groups, including esters (21 compounds), alcohols (four compounds), acids (four compounds), sesquiterpenes (two compounds) and aldehydes (one compound). Esters had the greatest contribution to the volatile composition in the case of every sample. The lowest relative abundance of esters was found in the case of the Gala variety cider fermented at 20 °C (G20), where they contributed to 58.81% total volatiles. The highest contribution of esters was found for the Cortland variety cider fermented at 15 °C (C15) and it equaled 79.58%. Ciders, fermented at 15 °C and 20 °C prepared with Konferencja variety juice, were characterized by higher contents of esters in the cases of apple varieties Idared and Gala (IK15, IK20, GK15, GK20) compared to the Idared and Gala ciders without pear addition (I15, I20, G15, G20). A different occurrence can be seen in the case of the Cortland variety cider. Ciders prepared from Cortland variety apple juice, fermented at 15 °C and 20 °C (C15, C20), are characterized by higher relative concentrations of esters than Cortland/Konferencja ciders (CK15, CK20). The alcohols constituted not only the second biggest group of compounds by the number of compounds identified, but also by their total relative abundance. The highest contribution of alcohols to the aroma profiles of ciders was found for G20, wherein they constituted 36.71% of total volatiles. The lowest contribution of alcohols to the aroma profile (16.07%) was found for C15. Addition of pear juice increased the amount of alcohols in Cortland variety ciders from 18.94% in C15 to 25.76% in CK15 and from 20.53% in C20 to 32.19% in CK20. In the cases of Gala and Idared cultivars, addition of pear juice had the opposite effect. Relative concentrations of alcohols decreased, from 24.6% for I15 to 19.9% for IK15; and from 22.45% for G15 to 19.43% for GK15. A similar trend could be noted for ciders fermented at 20 °C. The proportion of alcohols regarding volatile composition in I20 was equal to 29.04% and was smaller in IK20: 27.65%; in G20 alcohols composed 36.7% of volatiles, and in GK20 28.97%. Alcohol (except in GK20) was phenylethyl alcohol, which is produced by yeast and has a bread-like and flowery aroma [14]. Acids composed the third largest group of compounds and relative concentrations of these chemicals ranged from 3.16% in C15 to 9.18% in I15. A similar trend as in the case of alcohols could be seen in the ciders produced from mixed pear/apple juice. I15 and I20 were characterized by higher amounts of acids (9.18% and 8.29%, respectively) than IK15 and IK20 (4.27% and 6.36%); G15 and G20 also contained more acids in the volatile composition (4.7% and 4.15% respectively) than GK15 and GK20, which contained 3.28% and 3.67% of acids. Most prominent acid in all the ciders was octanoic acid, characterized by oily and waxy aroma [15]. Only one aldehyde, (E)-hex-2-enal, was found in the volatile compositions of ciders. It is a compound which has leafy aroma [16]. Its presence was not detected in four of the tested ciders (IK15, I20, C20, G20). (E)-hex-2-enal was found only in small quantities in other samples and constituted from 0.16% of volatiles in I15 to 0.96% in GK15. Its concentration was lower for the ciders prepared with pear juice addition only in the cases of IK15 and CK15. Sesquiterpenes were present in minuscule amounts in all tested ciders and their concentrations ranged from the lowest in GK20 (0.25% of all volatiles) to the highest in IK20 (0.83%) of all volatiles.

2.3. Carbohydrate and Ethanol Concentrations and Degree of Attenuation

The concentration of carbohydrates was determined in the juices prior to the fermentation by the means of high performance liquid chromatography (HPLC). Carbohydrate concentration has been presented as a sum of fermentable sugars (glucose, fructose and sucrose) (Table 3 and Table 4). The highest concentration of sugars in the juices was found for GK (13.9% w/w) and the lowest for I (10.9% w/w). At the end of fermentation, 14 days after inoculation with yeast, concentration of sugars was significantly lower and ranged from 2.23% (w/w) for I20 to 4.38% (w/w) for C15. The highest alcohol concentration was assessed in GC20 and was equal to 6.79% (v/v) and the lowest was determined in C15: 4.12% (v/v). The highest degree of fermentation was achieved by I20 and was equal 86.7%, and the lowest was achieved by C15, which equaled 60.54%.

3. Discussion

3.1. Volatile Compounds in Apple and Apple/Pear Juices

The aroma of apple juice has been the subject of much research and discussion [24,25,26]. Depending on the apple variety, climate and the use of different cultivation techniques, the researchers have pointed out significant differences between the aromatic profiles of analyzed juices. It has also been shown that differences in the production technology can have significant impacts on the aroma profiles of apple juices [27,28,29]. In this study, an attempt was made to show the differences in the quality of apple juice, which differ not only by the apple variety, such as Gala, Cortland and Idared, but to also assess how addition of pear Konferencja juice, can affect the volatile profile. Current knowledge about the formation of cider aromas resulting from mixtures of apple-pear juices is not sufficient and does not include a detailed analysis of the changes occurring in the volatile components of the juice building the aroma of cider which is formed during the fermentation process. Most researchers indicate methyl and ethyl esters of carboxylic acids as the main groups of compounds building the aroma of apple and pear juice [13,29,30,31,32,33]. In these studies, out of 46 marked chemical compounds which composed the aromas of apple and apple-pear juices, 32 compounds belonged to the group of esters. The main esters in the studies by Kato et al. [34], Aprea et al. [35] and Gasperi et al. [36] were: hexyl acetate, hexan-1-ol, butyl acetate and (Z)-hex-3-enyl acetate. In the tests performed, a large share of hexyl acetate (23–29%) was found as a compound shaping the aromatic profile of apple juice from the Idared and Gala varieties and their mixtures with pears (GK, IK). Other researchers, such as Xiaobo and Jiewen [37], who studied exotic Asian apples such as Fuji, Jina and Huaniu, also indicated the presence of the above-mentioned esters and a significant shares of, among others, propyl butanoate and propyl acetate. Other important groups of chemical compounds found in apple juices, which have been indicated by many studies, are alcohols ((E)-hex-2-en-1-ol, (Z)-hex-2-en-1-ol (Z), 2-methyl butan-1-ol, (Z)-hex-4-enol) and aldehydes ((E)-hex-2-enal, hexenal, nonanal, dodecanal) [29,32,33,34,35,36,37]. The research also showed significant influences of alcohols and aldehydes, such as (E)-hex-2-en-1-ol and (Z)-hex-2-en-1-ol, which dominated mainly in the juice of the Cortland variety, in building the aroma profiles of the analyzed juices. Other compound, belonging to the sesquiterpene group ((E, E)-α-farnesene) was found in the analyzed juices at a level close to the values presented by other researchers [38]. A relationship was also found between the content of farnesene isomers and the apple variety from which the juice originated. Ahn et al. [39] pointed to Cortland as the richest in the α-farnesene variety, placing apples of the Idared and Gala varieties behind it. Similar relations between the participation of farnesene in juices were also confirmed in this study. The addition of pear juice slightly decreased the share of farnesene in the overall aromatic profiles of the analyzed juices. Many researchers have shown that the aroma of pear juice is greatly dependent on the varietal factors, climate characteristics, cultivation techniques and degree of ripeness of the fruit used [13,40]. The main compounds characterized in the aromatic profiles of pears have included ethyl acetate, hexanal, ethyl butyrate and ethyl hexanoate. The addition of pear juice to apple juice affected the aromatic profile of the finished product. The main differences in the aromas of ciders are probably due to compounds such as ethyl-(2E,4Z)-deca-2,4-dienoate, which has a characteristic pear smell and is referred to by some researchers as a “pear ester” [13,41]. In the samples of mixed apple and pear juices, the percentage of this compound was significantly higher than in apple juices.

3.2. Volatile Compounds in Ciders

The amounts of volatile compounds contained in the tested cider samples depend on many variables. In addition to the varietal factor, fermentation temperature and types of microorganisms conducting the fermentation process, the analytical method used also plays a significant role in the amounts of the determined compounds (the main ones include GC, GC-MS, GC-MS/MS, GC-O, GC-MS-O) [10,42,43]. In this work, the most represented compounds identified in ciders included methyl and ethyl esters of carboxylic acids. The most important for building the aromatic profile were: ethyl hexanoate, ethyl acetate, 2-phenylethyl acetate, ethyl decanoate and ethyl dodecanoate. These compounds belong to the basic esters that build the aromas of apple and pear juices and are formed during fermentation processes with the use of Saccharomyces cerevisiae yeast [42,44,45]. Gamero et al., analyzing the effects of temperatures in the range 12–28 °C on the production of aromatic compounds by S. cerevisiae, used synthetic substrates to eliminate the effect of the matrix on the by-products of alcoholic fermentation [46]. During the research, it was found that at elevated fermentation temperature, S. cereviasie yeast produces increased amounts of esters such as ethyl acetate, 2-methylpropyl acetate, 3-methylbutyl acetate, hexyl acetate, ethyl hexanoate, ethyl octane and ethyl decanoate. Rolerro et al., while examining the effects of fermentation temperature and substrate supplementation on the shaping of the aromatic profiles of grape wines, found that the production of esters such as ethyl hexanoate or ethyl octanoate depend not only on the fermentation temperature, but also on the proportion of free nitrogen and fatty acids in the substrate [47]. In this work, a significant influence of fermentation temperature on the percentage share of esters in the aromatic profiles of the studied ciders was also observed. Fermentation temperature played a key role in the case of compounds such as ethyl hexanoate, ethyl octanoate and ethyl dodecanoate, whose percentages were higher in ciders fermented at 15 °C. Differences in observed relationships between fermentation temperature and the share of esters in the aromatic profiles of the analyzed ciders were due to differences between fermentation settings (contents of microelements, acidity or contents of extract). Ciders and cider musts are characterized by a much lower dry matter concentration, density, acidity and nitrogen content than wine musts [48]. Other important groups of compounds responsible for the cider aroma are alcohols and carboxylic acids. The research showed that the most important thing for shaping the aromatic profiles of the tested ciders from the Cortland, Idared and Gala juices were alcohols such as 3-methyl butan-1-ol, 2-methyl butan-1-ol, hexan-1-ol and phenylethyl alcohol, and acids such as octanoic and decanoic. Lobo et al., while analyzing Spanish ciders from Asturias and the Basque country have found alcohols such as butan-2-ol, propan-1-ol and hexan-1-ol, and acids such as hexanoic, octanoic and decanoic as dominant volatile compounds. The contents of these compounds in the cider production process increased significantly, which corresponds with the results obtained in this study [49]. Anton et al., analyzing the aromatic profiles of ciders using the yeast of the S. cerevisiae species and wild yeast (spontaneous fermentation) showed that the dominant alcohols in both cases were 2-methyl-butan-1-ol, 1-propan-1-ol, iso-butan-1-ol and 3-methylbutylalcohol and acids such as hexanoic, decanoic and octanoic [50]. Other researchers also point out important role of such volatiles as phenylethyl alcohol, whose aroma is described as honey-like and floral; hexyl alcohol, characterized by grassy notes; and alcohols such as 2-methyl-butan-1-ol and 3-methyl-butan-1-ol [51,52]. Sesquiterpenes are the group of compounds of great importance for the cider aroma, constituting 0.6% of all identified compounds. In this study, the content of compounds such as ((Z,E)-α-Farnesene, (E,E)-α-Farnesene in all tested cider musts) was determined. α-Farnesene isomers are often marked as volatile components in both apple and pear musts and ciders [38,53,54].
Current research in the field of cider production technology mainly concentrates on the contents of polyphenolic compounds and the analysis of aromatic profiles. These compounds play an important role due to their participation in the creation of the desired organoleptic characteristics and the bioactive and health-promoting properties of ciders. The current state of knowledge is also being expanded by research about issues related to the production of ciders using malo-lactic fermentation, used to obtain ciders with reduced acidity [55,56,57].

4. Materials and Methods

4.1. Raw Material

Juices from three apple cultivars: Idared, Cortland and Gala and pear cultivar: Konferencja were obtained from local orchard in Lubuskie voivodeship in village Solniki. Juices were pasteurized on PA1500 Pasteurizer (Voran Company, Pichl bei Wels, Austria) by juice producer “Słoneczny Sad” (Solniki, Poland) under the following conditions: 74 °C for 15 s under normal pressure.

4.2. Biological Material

Dry yeast used in this study to ferment apple and apple-pear musts to acquire cider was Saccharomyces cerevisiae Safdistill C-70 strain. It was obtained from the Fermentis division of Lesaffre company (Marqen Baroeul, France). Prior to inoculation, 2.5 g (dry weight) of dry yeast was rehydrated in 20 mL of sterile, distilled water and thermostated at 30 °C for 30 min with regular agitation.

4.3. Cider Production

Fermentation of apple juices and apple juice/pear juice mixes was carried out in 5.5 L glass vessels in a single repetition; 5 L of juice was inoculated by 20 mL of yeast solution. The fermentation was carried out for 14 days. Six cider variants were fermented at 15 °C: 100% Idared apple juice (I15); 100% Cortland apple juice (C15); 100% Gala apple juice (G15); 80% Idared apple juice and 20% Konferencja pear juice mix (IK15); 80% Cortland apple juice and 20% Konferencja pear juice mix (IK15); 80% Gala apple juice and 20% Konferencja pear juice mix (GK15). Six cider variants were fermented at 20°C: 100% Idared apple juice (I20); 100% Cortland apple juice (C20); 100% Gala apple juice (G20); 80% Idared apple juice and 20% Konferencja pear juice mix (IK20); 80% Cortland apple juice and 20% Konferencja pear juice mix (IK20); 80% Gala apple juice and 20% Konferencja pear juice mix (GK20). Ciders, after fermentation, were bottled and kept at 4 °C.

4.4. Volatile Compounds Analysis

Solid-phase microextraction (SPME) techniques were applied for the isolation and concentration of volatile flavor compounds [58]. A representative cider sample (40 mL) was transferred to a 100 mL septum-closed vials with 0.5 µg 2-undecanone added as an internal standard for quantification. Samples were held in an 80 °C water bath and shaken during the isolation. Each sample was allowed to equilibrate and absorb onto the SPME fiber (2 cm–50/30 µm DVB/CAR/PDMS; Supelco, Inc., Bellefonte, PA, USA) for 45 min. A gas chromatograph Clarus 680 coupled with a mass spectrometry detector (GC-MS) SQ 8S by Perkin Elmer, with a column ELITE-5MS (Crossbond 5% diphenyl and 95% dimethylpolysiloxane) 30 m × 0.25 mm ID × 0.25 μm film. Scanning was performed from 50 to 300 m/z in electron impact (EI) at 70 eV, mode at 5 scan s−1. Analyses were carried out using helium as carrier gas at a flow rate of 1.0 mL min−1 in a split ratio 1:10 and the following temperature program: (a) 40 °C for 3 min; (b) rate of 5.0 °C min−1 from 40 to 160 °C; (c) rate of 30 °C min−1 from 160 to 280 °C. Injector was held constantly at 220 °C. Compounds were identified by using 3 different analytical methods: (1) retention indices (KI), (2) GC-MS retention times (authentic chemicals) and (3) comparison of mass spectra, with database (NIST 11 Mass Spectral and Retention Index Libraries (The National Institute of Standards and Technology, Gaithersburg, MD, USA) with similarity indexes set above 90%. For comparison of obtained mass spectra Automated Mass Spectral Deconvolution and Identification System (AMDIS) software version 2.73 was used.
Linear retention indices were calculated against C6-C30 n-alkanes mix (Sigma-Aldrich, Saint Louis, MO, USA).
Quantification was based on the peak area normalization. For data processing, ACD/Spectrus Processor 2017.2.1 software (Advanced Chemistry Development, Inc. Toronto, Ontario M5C 1B5, Canada) was used.

4.5. Carbohydrates and Ethanol Concentration and Degree of Attenuation

Concentration of carbohydrates (sucrose, glucose and fructose) in ciders, apple and pear juices and ethanol content of ciders was assessed by the means of high performance liquid chromatography (HPLC). Samples of cider and juice were 3-fold diluted with de-ionized water and filtered through 0.2 µm nylon membrane filter. Analyses were performed using Shimadzu Prominence chromatograph (Shimadzu Corp., Tokyo, Japan) equipped with Rezex ROA-Organic Acid H+ column (300 × 7.8 mm) (Phenomenex, Torrance, USA). The samples were loaded into 20 µL injection loop and eluted at 50 °C with 0.005 M H2SO4 as a mobile phase at a flow rate of 0.4 mL min−1. The compounds were detected using Shimadzu RID-10A refractive index detector; detection temperature was maintained at 50 °C. Integration and quantification of chromatograms were performed using Chromax 10 software (Pol-Lab, Warsaw, Poland). Degree of attenuation was calculated using the formula below:
(SJ − SC)/SJ = Degree of attenuation
SJ-carbohydrate content in juice
SC-carbohydrate content in finished cider

4.6. Statistical Analysis

The results of volatile compounds concentration analysis and sugars and ethanol content were statistically analyzed using Duncan’s test at the significance level of α = 0.05. In the volatile analysis, three-way analysis of variance (ANOVA) was performed to detect significant differences in the analyte concentrations depending on the main factors studied (fermentation temperature, cultivar of apples and addition of pear juice). Statistica software in version 10 was used for data processing.

5. Conclusions

The sensory qualities of cider are linked to the volatile compounds—which are composed of volatiles present in the raw material used for cider-making—and to the metabolites created by the microorganisms during the fermentation process. In this study, 46 volatiles have been identified in the pear juice/apple juice mixes and 33 volatiles have been found in the ciders prepared from these juices. The main chemical compounds found in Idared apple juice were 2-methyl ethyl butanoate, hexyl acetate and 2-methylpropyl acetate. The main aromatic constituents in the volatilome of Cortland apple variety juice were (E)-hex-2-en-1-ol, 2-methylpropyl acetate and (Z)-hex-2-en-1-ol (Z). In the composition of volatile profile of Gala juice, (E)-hex-2-enol, hexyl acetate and ethyl oct-2-enoate were recognized as the most abundant constituents. The addition of pear juice added esters with a characteristic pear aroma, such as methyl (2E,4Z)-deca-2,4-dienoate and ethyl (2E, 4Z)-deca-2,4-dienoate. Fermentation of juices changed composition of the juices’ volatilomes. In the ciders, the main chemical components were esters. The most abundant esters determined in all of the ciders were ethyl decanoate and ethyl octanoate. In the ciders prepared from juice with pear juice addition, ethyl (2E,4Z)-deca-4-dienoate, characteristic of the pear aroma, was found. Ciders were characterized by a higher degree of alcohols, such as hexan-1-ol, phenylethyl alcohol and 2-methyl butan-1-ol, in the volatile profiles than in juices from which they were prepared. Ciders were also characterized by lower relative abundances of sesquiterpenes, such as (Z,E)-α-farnesene and (E,E)-α-farnesene, in the volatilome than in apple and pear juices.

Author Contributions

Conceptualization, J.K. and J.K.-R.; methodology, J.K., J.K.-R. and A.S.; software, J.K. and A.S.; validation, J.K., J.K.-R. and A.S.; formal analysis, J.K. and J.K.-R.; investigation, J.K.; resources, J.K., J.K.-R. and A.S.; data curation, J.K., J.K.-R. and A.S.; writing—original draft preparation, J.K, A.G. (Alan Gasiński); writing—review and editing, J.K., A.G. (Alan Gasiński) and A.G. (Adam Głowacki); visualization, J.K. and A.G. (Alan Gasiński); supervision, J.K.-R.; project administration, J.K.-R. and A.S.; funding acquisition, J.K.-R. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Coton, E.; Coton, H.; Guichard, H. Cider (Cyder; Hard Cider): The Product and Its Manufacture. In Encyclopedia of Food and Health, 1st ed.; Cabalerro, B., Finglas, P.M., Toldrá, F., Eds.; Academic Press: Oxford, UK, 2016; Volume 2, pp. 119–128. [Google Scholar]
  2. Dz. U. 2011 Nr 120 poz. 690. Ustawa z dnia 12 maja 2011r. o wyrobie i rozlewie wyrobów winiarskich, obrocie tymi wyrobami i organizacji rynku wina.
  3. Slavin, J.L.; Lloyd, B. Health benefits of fruits and vegetables. Adv. Nutr. An Int. Rev. J. 2012, 3, 506–516. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Wang, X.; Ouyang, Y.; Liu, J.; Zhu, M.; Zhao, G.; Bao, W.; Hu, F.B. Fruit and vegetable consumption and mortality from all causes, cardiovascular disease, and cancer: Systematic review and dose-response meta-analysis of prospective cohort studies. BMJ (Clin. Res. Ed.) 2014, 349, 4490. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Reiland, H.; Slavin, J. Systematic Review of Pears and Health. Nutr. Today 2015, 50, 301–305. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  6. Qin, Z.H.; Petersen, M.A.; Bredie, W.L.P. Flavor profiling of apple ciders from the UK and Scandinavian region. Food Res. Int. 2018, 105, 713–723. [Google Scholar] [CrossRef]
  7. Symoneaux, R.; Guichard, H.; LeQuere, J.M.; Baron, A.; Chollet, S. Could cider aroma modify cider mouthfeel properties? Food Qual. Prefer. 2015, 45, 11–17. [Google Scholar] [CrossRef]
  8. Plutowska, B.; Wardenecki, W. Gas chromatography-olfactometry of alcoholic beverages. In Alcoholic Beverages. Sensory Evaluation and Consumer Research, 1st ed.; Piggot, J., Ed.; Woodhead Publishing: Cambridge, UK, 2011; pp. 101–130. [Google Scholar]
  9. Alberti, A.; Dos Santos, T.P.M.; Zielinski, A.A.F.; Dos Santos, C.M.E.; Braga, C.M.; Demiate, I.M.; Nogueira, A. Impact on chemical profile in apple juice and cider made from unripe, ripe and senescent dessert varieties. LWT 2016, 65, 436–443. [Google Scholar] [CrossRef]
  10. Cousin, F.J.; Le Guellec, R.; Schlusselhuber, M.; Dalmasso, M.; Laplace, J.-M.; Cretenet, M. Microorganisms in Fermented Apple Beverages: Current Knowledge and Future Directions. Microorganisms 2017, 5, 39. [Google Scholar] [CrossRef] [Green Version]
  11. Williams, A.A. The development of a vocabulary and profile assessment method for evaluating the flavour contribution of cider and perry aroma constituents. J. Sci. 1975, 26, 567–582. [Google Scholar] [CrossRef]
  12. Niu, Y.; Liu, Y.; Xiao, Z. Evaluation of Perceptual Interactions between Ester Aroma Components in Langjiu by GC-MS, GC-O, Sensory Analysis, and Vector Model. Foods 2020, 9, 183. [Google Scholar] [CrossRef] [Green Version]
  13. Wang, C.; Zhang, W.; Li, H.; Mao, J.; Guo, C.; Ding, R.; Wang, Y.; Fang, L.; Chen, Z.; Yang, G. Analysis of Volatile Compounds in Pears by HS-SPME-GC×GC-TOFMS. Molecules 2019, 24, 1795. [Google Scholar] [CrossRef] [Green Version]
  14. Gamero, A.; Dijkstra, A.; Smit, B.; de Jong, C. Aromatic Potential of Diverse Non-Conventional Yeast Species for Winemaking and Brewing. Fermentation 2020, 6, 50. [Google Scholar] [CrossRef]
  15. Liu, R.; Wang, C.; Huang, A.; Lv, B. Characterization of Odors of Wood by Gas Chromatography-Olfactometry with Removal of Extractives as Attempt to Control Indoor Air Quality. Molecules 2018, 23, 203. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. El Youssef, C.; Bonnarme, P.; Fraud, S.; Péron, A.-C.; Helinck, S.; Landaud, S. Sensory Improvement of a Pea Protein-Based Product Using Microbial Co-Cultures of Lactic Acid Bacteria and Yeasts. Foods 2020, 9, 349. [Google Scholar] [CrossRef] [Green Version]
  17. Arcari, S.G.; Caliari, V.; Sganzerla, M.; Godoy, H.T. Volatile composition of Merlot red wine and its contribution to the aroma: Optimization and validation of analytical method. Talanta 2017, 174, 752–766. [Google Scholar] [CrossRef] [PubMed]
  18. Ruiz, M.J.; Zea, L.; Moyano, L.; Medina, M. Aroma active compounds during the drying of grapes cv. Pedro Ximenez destined to the production of sweet Sherry wine. Eur. Food Res. Technol. 2010, 230, 429–435. [Google Scholar] [CrossRef]
  19. Czerny, M.; Christlbauer, M.; Fischer, A.; Granvogl, M.; Hammer, M.; Hartl, C.; Hernandez, N.; Schieberle, P. Re-investigation on odour thresholds of key food aroma compounds and development of an aroma language based on odour qualities of defined aqueous odorant solutions. Eur. Food Res. Technol. 2008, 228, 265–273. [Google Scholar] [CrossRef]
  20. Englezos, V.; Torchio, F.; Cravero, F.; Marengo, F.; Giacosa, S.; Gerbi, V.; Rantsiou, K.; Rolle, L.; Cocolin, L. Aroma profile and composition of Barbera wines obtained by mixed fermentations of Starmerella bacillaris (synonym Candida zemplinina) and Saccharomyces cerevisiae. Lwt-Food Sci. Technol. 2016, 73, 567–575. [Google Scholar] [CrossRef]
  21. Moreno-Garcia, J.; Garcia-Martinez, T.; Millan, M.C.; Mauricio, J.C.; Moreno, J. Proteins involved in wine aroma compounds metabolism by a Saccharomyces cerevisiae flor-velum yeast strain grown in two conditions. Food Microbiol. 2015, 51, 1–9. [Google Scholar] [CrossRef]
  22. Xiao, Z.B.; Luo, J.; Niu, Y.W.; Wang, P.P.; Wang, R.L.; Sun, X.X. Olfactory impact of esters on rose essential oil floral alcohol aroma expression in model solution. Food Res. Int. 2019, 116, 211–222. [Google Scholar] [CrossRef]
  23. Condurso, C.; Cincotta, F.; Tripodi, G.; Sparacio, A.; Giglio, D.M.L.; Sparla, S.; Verzera, A. Effects of cluster thinning on wine quality of Syrah cultivar (Vitis vinifera L.). Eur. Food Res. Technol. 2016, 242, 1719–1726. [Google Scholar] [CrossRef]
  24. Bingman, M.T.; Stellick, C.E.; Pelkey, J.P.; Scott, J.M.; Cole, C.A. Monitoring Cider Aroma Development throughout the Fermentation Process by Headspace Solid Phase Microextraction (HS-SPME) Gas Chromatography–Mass Spectrometry (GC-MS) Analysis. Beverages 2020, 6, 40. [Google Scholar] [CrossRef]
  25. Nicolini, G.; Román, T.; Carlin, S.; Malacarne, M.; Nardin, T.; Bertoldi, D.; Larcher, R. Characterisation of singlevariety still ciders produced with dessert apples in the Italian Alps. J. Inst. Brew. 2018, 124, 457–466. [Google Scholar]
  26. Verdu, C.; Childebrand, N.; Marnet, N.; Lebail, G.; Dupuis, F.; Laurens, F.; Guilet, D.; Guyot, S. Polyphenol Variability In The Fruits Additionally, Juices Of A Cider Apple Progeny. J. Sci. Food Agric. 2014, 94, 7. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  27. Rosend, J.; Kuldjärv, R.; Rosenvald, S.; Paalme, T. The effects of apple variety, ripening stage, and yeast strain on the volatile composition of apple cider. Heliyon 2019, 5, e01953. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  28. Schmutzer, G.R.; Magdas, A.D.; Leontin, I.D.; Moldovan, Z. Determination of the Volatile Components of Apple Juice Using Solid Phase Microextraction and Gas Chromatography–Mass Spectrometry. Anal. Lett. 2014, 47, 1683–1696. [Google Scholar] [CrossRef]
  29. Jing, C.; Feng, D.; Zhao, Z.; Wu, X.; Chen, X. Effect of environmental factors on skin pigmentation and taste in three apple cultivars. Acta Physiol. Plant. 2020, 42, 1–12. [Google Scholar] [CrossRef]
  30. Mehinagic, E.; Royer, G.; Symoneaux, R.; Jourjon, F.; Prost, C. Characterization of Odor-Active Volatiles in Apples: Influence of Cultivars and Maturity Stage. J. Agric. Food Chem. 2006, 54, 2678–2687. [Google Scholar] [CrossRef]
  31. Holland, D.; Larkov, O.; Bar-Ya’Akov, I.; Bar, E.; Zax, A.; Brandeis, E.; Ravid, U.; Lewinsohn, E. Developmental and Varietal Differences in Volatile Ester Formation and Acetyl-CoA: Alcohol Acetyl Transferase Activities in Apple (Malus domestica Borkh.) Fruit. J. Agric. Food Chem. 2005, 53, 7198–7203. [Google Scholar] [CrossRef]
  32. Kebede, B.; Ting, V.; Eyres, G.; Oey, I. Volatile Changes during Storage of Shelf Stable Apple Juice: Integrating GC-MS Fingerprinting and Chemometrics. Foods 2020, 9, 165. [Google Scholar] [CrossRef] [Green Version]
  33. Both, V.; Thewes, F.B.; Brackmann, A.; Ferreiran, D.; Pavanellon, E.P.; Wagner, R. Effect of low oxygen conditioning and ultralow oxygen storage on the volatile profile, ethylene production and respiration rate of ‘Royal Gala’ apples. Sci. Hortic. 2016, 2009, 156. [Google Scholar] [CrossRef]
  34. Kato, T.; Shimoda, M.; Suzuki, J.; Kawaraya, A.; Igura, N.; Hayakawa, I. Changes in the odors of squeezed apple juice during thermal processing. Food Res. Int. 2003, 36, 777–785. [Google Scholar] [CrossRef]
  35. Aprea, E.; Charles, M.; Endrizzi, I.; Corollaro, L.; Betta, M.; Biasioli, E.; Gasperi, F. Sweet taste in apple: The role of sorbitol, individual sugars, organic acids and volatile compounds. Sci. Rep. 2017, 7, 44950. [Google Scholar] [CrossRef] [PubMed]
  36. Gasperi, F.; Aprea, E.; Biasioli, F.; Carlin, S.; Endrizzi, I.; Pirretti, G.; Spilimbergo, S. Effects of supercritical CO2 and N2O pasteurisation on the quality of fresh apple juice. Food Chem. 2009, 115, 129–136. [Google Scholar] [CrossRef]
  37. Xiaobo, Z.; Jiewen, Z. Comparative analyses of apple aroma by a tin-oxide gas sensor array device and GC/MS. Food Chem. 2008, 107, 120–128. [Google Scholar] [CrossRef]
  38. Tsantili, E.; Gapper, N.E.; Apollo Arquiza, J.M.R.; Whitaker, B.D.; Watkins, C.B. Ethylene and α-Farnesene Metabolism in Green and Red Skin of Three Apple Cultivars in Response to 1-Methylcyclopropene (1-MCP) Treatment. J. Agric. Food Chem. 2007, 55, 5267–5276. [Google Scholar] [CrossRef] [PubMed]
  39. Ahn, T.; Paliyath, G.; Murr, D.P. Antioxidant enzyme activities in apple varieties and resistance to superficial scald development. Food Res. Int. 2007, 40, 1012–1019. [Google Scholar] [CrossRef]
  40. Qin, G.; Tao, S.; Zhang, H.; Huang, W.; Wu, J.; Xu, Y.; Zhang, S. Evolution of the Aroma Volatiles of Pear Fruits Supplemented with Fatty Acid Metabolic Precursors. Molecules 2014, 19, 20183–20196. [Google Scholar] [CrossRef] [Green Version]
  41. Bengtsson, J.; Gonzalez, F.; Cattaneo, A.M.; Montagné, N.; Walker, W.; Bengtsson, M.; Anfora, G.; Ignell, R.; Jacquin-Joly, E.; Witzgall, P. A predicted sex pheromone receptor of codling moth Cydia pomonella detects the plant volatile pear ester. Front Ecol. Evol. 2014, 2, 1–11. [Google Scholar] [CrossRef] [Green Version]
  42. Haider, W.; Barillier, D.; Hayat, A.; Gaillard, J.; Ledauphin, J. Rapid quantification and comparison of major volatile compounds of ciders from France (Normandy and Brittany) using microextraction by packed sorbent (MEPS). Anal. Methods 2014, 6, 1364–1376. [Google Scholar] [CrossRef]
  43. Domingo-Almenara, X.; Brezmes, J.; Vinaixa, M.; Samino, S.; Diaz, M.; Ibanez, L.; Correig, X.; Perera, A.; Yanes, O. eRah: A computational tool integrating spectral deconvolution and alignment with quantification and identification of metabolites in GC-MS-based metabolomics. Anal. Chem. 2016, 88, 9821–9829. [Google Scholar] [CrossRef]
  44. Zhu, J.; Shi, J.; Lu, Y.; Liu, L.; Liu, Y. Application of strains of Geotrichum spp. to decrease higher alcohols and to increase esters. J. Inst. Brew. 2016, 122, 147–155. [Google Scholar]
  45. Liu, S.; Aung, M.T.; Lee, P.; Yu, B. Yeast and volatile evolution in cider co-fermentation with Saccharomyces cerevisiae and Williopsis saturnus. Ann. Microbiol. 2016, 66, 307–315. [Google Scholar] [CrossRef]
  46. Gamero, A.; Tronchoni, J.; Querol, A.; Belloch, C. Production of aroma compounds by cryotolerant Saccharomyces species and hybrids at low and moderate fermentation temperatures. J. Appl. Microbiol. 2013, 114, 1405–1414. [Google Scholar] [CrossRef] [PubMed]
  47. Rollero, S.; Bloem, A.; Camarasa, C.; Sanchez, I.; Ortiz-Julien, A.; Sablayrolles, J.M.; Dequin, S.; Mouret, J.R. Combined effects of nutrients and temperature on the production of fermentative aromas by Saccharomyces cerevisiae during wine fermentation. Appl. Microbiol. Biot. 2015, 99, 291–304. [Google Scholar] [CrossRef]
  48. Dierings, L.R.; Braga, C.M.; Silva, K.M.; Wosiacki, G.; Nogueira, A. Population dynamics of mixed cultures of yeast and lactic acid bacteria in cider conditions. Braz. Arch. Biol. Technol. 2013, 56, 837–847. [Google Scholar] [CrossRef] [Green Version]
  49. Lobo, A.P.; Antón-Díaz, M.J.; Mangas-Alonso, J.J.; Suárez Valles, B. Characterization of Spanish ciders by means of chemical and olfactometric profiles and chemometrics. Food Chem. 2016, 213, 505–513. [Google Scholar] [CrossRef]
  50. Antón-Díaz, M.J.; Suárez Valles, B.; Mangas-Alonso, J.J.; Fernández-García, O.; Lobo, A.P. Impact of different techniques involving contact with lees on the volatile composition of cider. Food Chem. 2016, 190, 1116–1122. [Google Scholar] [CrossRef]
  51. Liu, P.; Lu, L.; Duan, C.; Yan, G. The contribution of indigenous non-Saccharomyces wine yeast to improved aromatic quality of Cabernet Sauvignon wines by spontaneous fermentation. LWT-Food Sci. Technol. 2016, 71, 356–363. [Google Scholar] [CrossRef]
  52. Spinosa, W.A.; dos Júnior, V.S.; Galvan, D.; Fiorio, J.L.; Gomez, R.J.H.C. Fermentation kinetics of rice syrup, with high contente of dextrose equivalente, by Saccharomyces cerevisae and characterization of volatile compounds from wine. J. Food Process. Pres. 2016, 40, 1199–1205. [Google Scholar] [CrossRef]
  53. Versini, G.; Franco, M.A.; Moser, S.; Manca, G. Characterisation of pear distillates from wild and cultivated varieties in Sardinia. Int. J. Food Tech. 2012, 47, 2519–2531. [Google Scholar] [CrossRef]
  54. Villière, A.; Arvisenet, G.; Lethuaut, L.; Prost, C.; Sérot, T. Selection of a representative extraction method for the analysis of odourant volatile composition of French cider by GC-MS-O and GCxGC-TOF-MS. Food Chem. 2012, 131, 1561–1568. [Google Scholar] [CrossRef]
  55. Lachowicz, S.; Oszmiański, J.; Uździcka, M.; Chmielewska, J. The Influence of Yeast Strain, β-Cyclodextrin, and Storage Time on Concentrations of Phytochemical Components, Sensory Attributes, and Antioxidative Activity of Novel Red Apple Ciders. Molecules 2019, 24, 2477. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  56. Al Daccache, M.; Koubaa, M.; Maroun, R.G.; Salameh, D.; Louka, N.; Vorobiev, E. Suitability of the Lebanese “Ace Spur” Apple Variety for Cider Production Using Hanseniaspora sp. Yeast. Fermentation 2020, 6, 32. [Google Scholar] [CrossRef] [Green Version]
  57. Nešpor, J.; Karabín, M.; Štulíková, K.; Dostálek, P. An HS-SPME-GC-MS Method for Profiling Volatile Compounds as Related to Technology Used in Cider Production. Molecules 2019, 24, 2117. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  58. Boylston, T.D.; Wang, H.; Reitmeier, C.A.; Glatz, B. Effects of Processing Treatment and Sorbate Addition on the Flavor Characteristics of Apple Cider. J. Agric. Food Chem. 2003, 51, 1924–1931. [Google Scholar] [CrossRef]
Table 1. Volatile compounds identified in analyzed juices.
Table 1. Volatile compounds identified in analyzed juices.
CompoundAroma Descriptor 1Retention IndicesChemical FamilyIIKCCKGGK 2
Exp.Lit. [%] of Total Peak Area
Butan-1-olfruity, medicinal674675Alcohols0.72 ± 0.22b0.01 ± 0.01c0.07 ± 0.0c0.01 ± 0.01c2.31 ± 1.15a0.09 ± 0.05c
Propyl acetatesweet, fruity705708Esters0.01 ± 0.01d0.59 ± 0.23a0.06 ± 0.04bc0.08 ± 0.05bc0.10 ± 0.06b0.02 ± 0.02d
2-methylbutan-1-olwiney, oniony,740745Alcohols3.97 ± 1.98a3.09 ± 1.84b2.28 ± 1.04c2.34 ± 1.23c0.80 ± 0.53d0.36 ± 0.24e
Butyl acetateappley812812Esters0.68 ± 0.44a0.49 ± 0.32b0.05 ± 0.04d0.01 ± 0.01d0.15 ± 0.08c0.05 ± 0.03d
(E)-hex-2-en-1-olleafy, fresh832840Alcohols0.42 ± 0.28d0.38 ± 0.21d13.00 ± 5.42b18.30 ± 6.08a3.98 ± 2.15c3.42 ± 1.78c
Propan-2-yl butanoate banana-like843843Esters0.10 ± 0.06b0.05 ± 0.03c0.45 ± 0.31a0.16 ± 0.09b0.11 ± 0.06b0.07 ± 0.04c
Ethyl 2-methylbutanoateappley846845Esters19.50 ± 4.38a19.00 ± 5.02a1.62 ± 0.76d2.43 ± 1.15c5.62 ± 2.44b5.67 ± 2.83b
(E)-hex-2-enalleafy849847Aldehydes0.01 ± 0.01d1.30 ± 0.95b0.37 ± 0.24c0.05 ± 0.03d0.07 ± 0.04d10.10 ± 4.32a
(E)-hex-2-en-1-olleafy, winey, appley855854Alcohols1.70 ± 1.05d1.04 ± 0.82d43.10 ± 8.53a43.40 ± 9.12a10.20 ± 6.41b6.63 ± 2.81c
Hexan-1-olflowery865863Alcohols1.26 ± 0.74a0.01 ± 0.01d0.70 ± 0.53b0.01 ± 0.01d0.20 ± 0.13c0.01 ± 0.01d
(Z)-hex-4-en-1-olfruity, fresh,873870Alcohols5.40 ± 3.11a4.17 ± 2.83b0.01 ± 0.01d0.06 ± 0.04d4.64 ± 2.72ab2.39 ± 1.42c
2-methylpropyl acetatesweet, banana-like883883Esters25.10 ± 9.42a25.60 ± 10.83a6.47 ± 4.18bc5.53 ± 2.78d7.64 ± 4.72b6.18 ± 2.81cd
Propyl butanoatefruity,904898Esters0.01 ± 0.01e0.20 ± 0.12bc0.09 ± 0.05d0.33 ± 0.18a0.16 ± 0.09c0.13 ± 0.07c
Butyl propanoateearthy, sweet916915Esters0.34 ± 0.21e0.65 ± 0.29d1.72 ± 0.94ab1.56 ± 0.82b1.75 ± 0.97a1.23 ± 0.72bc
Pentyl acetatebanana-like, etheric. fruity923920Esters0.92 ± 0.42c1.99 ± 1.05a0.55 ± 0.34d0.14 ± 0.08e0.85 ± 0.42c1.68 ± 1.12b
Tert-butyl butanoatefruity, sweet, appley955956Esters0.14 ± 0.08c0.13 ± 0.09c1.18 ± 0.66a1.10 ± 0.54a0.19 ± 0.12bc0.13 ± 0.08c
2-methylbutyl propanoatefruity, etheric963965Esters0.03 ± 0.02c0.28 ± 0.16a0.06 ± 0.04b0.06 ± 0.05b0.02 ± 0.02c0.03 ± 0.02c
Benzaldehydeoleic, nutty, sour cherry970960Aldehydes0.03 ± 0.02d0.07 ± 0.05d0.35 ± 0.18a0.15 ± 0.09c0.15 ± 0.11c0.22 ± 0.14bc
Heptan-1-olleafy, musty983980Alcohols0.10 ± 0.06bc0.25 ± 0.16a0.31 ± 0.23a0.06 ± 0.04c0.05 ± 0.04c0.01 ± 0.01c
Butyl butanoatesweet, fruity, fresh1001998Esters0.60 ± 0.27c1.98 ± 0.82a0.11 ± 0.06d1.33 ± 0.78b1.16 ± 0.72b1.76 ± 1.02a
hex-5-enyl acetatesweet, leafy, fresh10111010Esters0.10 ± 0.06b0.08 ± 0.05b0.09 ± 0.05b0.15 ± 0.09a0.15 ± 0.06a0.17 ± 0.08a
Hexyl acetatesweet, green apple, banana10221022Esters29.80 ± 6.72a29.80 ± 8.44a5.55 ± 2.31c5.30 ± 2.12c23.30 ± 7.12b28.60 ± 5.15a
Ethyl hex-2-enoatesweet, fresh, leafy,10381040Esters0.35 ± 0.18a0.06 ± 0.04c0.07 ± 0.03c0.13 ± 0.06b0.05 ± 0.04c0.10 ± 0.07bc
Butyl 2-methylbutanoatefruity, coconutty,10501045Esters0.68 ± 0.32c0.58 ± 0.27c0.37 ± 0.14d0.30 ± 0.16d2.56 ± 1.82a2.02 ± 1.04b
Butyl 3-methylbutanoateapple skin10671070Esters0.05 ± 0.03d0.28 ± 0.12ab0.02 ± 0.02d0.23 ± 0.11b0.15 ± 0.08c0.12 ± 0.07c
Octan-1-olcitrusy, flowery, aldehydy10821080Alcohols0.42 ± 0.28c0.32 ± 0.20d0.84 ± 0.44ab0.80 ± 0.38b0.91 ± 0.47a0.42 ± 0.31c
Pentyl butanoatepineappley, banana, tropical10991098Esters0.14 ± 0.09c0.15 ± 0.06c0.97 ± 0.58a0.73 ± 0.42b0.17 ± 0.12c0.20 ± 0.14c
Hexyl propanoatefruity, green pear11111114Esters0.05 ± 0.03e0.02 ± 0.02e0.40 ± 0.28c0.16 ± 0.12d2.03 ± 1.05a1.31 ± 0.72b
Heptyl acetatelemongrass11181118Esters0.05 ± 0.04d0.06 ± 0.04d0.45 ± 0.22a0.30 ± 0.18b0.10 ± 0.08c0.11 ± 0.07c
2-methylpentyl butanoateappley, fruity11451150Esters0.14 ± 0.08a0.12 ± 0.08a0.14 ± 0.06a0.11 ± 0.07ab0.09 ± 0.05b0.15 ± 0.12a
2-methylhexyl propanoatefruity, appley, sweet11541155Esters0.59 ± 0.33a0.38 ± 0.23b0.16 ± 0.07cd0.20 ± 0.14c0.10 ± 0.06d0.09 ± 0.06d
Octan-1-olwoody, herbal, fresh11741180Alcohols0.31 ± 0.22b0.41 ± 0.28a0.07 ± 0.04d0.11 ± 0.06cd0.04 ± 0.03e0.01 ± 0.01e
Butyl hexanoatepineappley, waxy, fruity11961198Esters0.30 ± 0.18d0.29 ± 0.11d8.86 ± 3.12a6.62 ± 2.83bc5.30 ± 2.41c5.25 ± 2.62c
Ethyl (Z)-oct-2-enoatetropical, waxy, fruity,12001202Esters0.36 ± 0.28c0.45 ± 0.31c0.19 ± 0.08d0.44 ± 0.28c12.30 ± 2.34a10.50 ± 3.12b
Octyl acetatemushroom-like, herbal, earthy12161212Esters0.65 ± 0.36a0.13 ± 0.07b0.07 ± 0.05c0.09 ± 0.06bc0.10 ± 0.06b0.11 ± 0.08b
2,4-dimethylobenzaldehydechemical, vanilla, sour cherry,12221230Aldehydes0.31 ± 0.18a0.02 ± 0.02d0.06 ± 0.05cd0.08 ± 0.04c0.15 ± 0.10b0.03 ± 0.02d
Decanoic acidappley, fruity, banana12421245Acids2.41 ± 1.63e2.83 ± 1.82d4.30 ± 2.93c2.64 ± 1.44de9.28 ± 2.55a8.59 ± 3.12b
3-methylbutyl hexanoatepineappley, fruity12581259Esters0.11 ± 0.08bc0.05 ± 0.04d0.01 ± 0.01d0.08 ± 0.02c0.07 ± 0.04cd0.46 ± 0.28a
(Z)-dec-3-en-1-olwaxy, fruity12661275Alcohols0.05 ± 0.04c0.03 ± 0.02c0.19 ± 0.12a0.08 ± 0.06b0.09 ± 0.05b0.04 ± 0.03c
Heptyl 2-methylbutanoateappley13391342Esters0.06 ± 0.04c0.06 ± 0.03c0.16 ± 0.12a0.10 ± 0.04b0.09 ± 0.05b0.02 ± 0.02d
Ethyl (Z)-dec-4-enoategrapefruity, waxy, fruity, appley13611364Esters0.00b0.01 ± 0.01a0.00b0.01 ± 0.01a0.00b0.01 ± 0.01a
Methyl (2E,4Z)-deca-2,4-dienoatepear13861383Esters0.00b0.02 ± 0.01a0.00b0.04 ± 0.02a0.00b0.03 ± 0.02a
Hexyl hexanoatetropical, waxy, fruity13891385Esters0.06 ± 0.04d0.78 ± 0.42b0.07 ± 0.04d0.80 ± 0.38b1.23 ± 0.67a0.15 ± 0.11c
Ethyl (2E,4Z)-deca-2,4-dienoatetropical, pear14791474Esters0.00b0.09 ± 0.06a0.00b0.11 ± 0.07a0.00b0.08 ± 0.05a
(Z,E)-α-Farnesenewoody14911495Sesquiterpenes0.23 ± 0.11b0.21 ± 0.12b0.31 ± 0.18a0.33 ± 0.19a0.11 ± 0.08c0.09 ± 0.07c
(E,E)-α-Farnesenewoody, citrusy15071505Sesquiterpenes1.73 ± 0.98c1.48 ± 0.82d4.06 ± 2.02a2.95 ± 1.62b1.49 ± 1.12d0.98 ± 0.72e
1 [17,18,19,20,21,22,23]; 2 I—Idared apple variety juice; IK—Idared apple (80%) and Konferencja pear (20%) juices mix; C—Cortland apple variety juice; CK-+—Cortland apple (80%) and Konferencja pear (20%) juices mix; G—Gala apple variety juice; GK—Idared apple (80%) and Konferencja pear (20%) juices mix. Values are expressed as means (n = 2) ± standard deviations. Mean values with different letters (a, b, c, d, e) within the same column are statistically different (p-value < 0.05).
Table 2. Volatile compounds identified in analyzed ciders.
Table 2. Volatile compounds identified in analyzed ciders.
CompoundAroma Descriptor 1Retention IndicesChemical FamilyI15IK15C15CK15 G15GK15I20IK20C20CK20 G20GK20 2
Exp.Lit. [%] of Total Peak Area
3-methylbutan-1-olfusel, alcoholic699705Alcohols6.49 ± 1.98c4.19 ± 1.42e7.11 ± 2.77bc8.71 ± 2.34b9.98 ± 2.18a7.96 ± 3.02b5.36 ± 1.04d4.67 ± 1.88e4.16 ± 1.27e4.96 ± 2.21de9.09 ± 2.66ab7.82 ± 2.68b
2-methylbutan-1-olwiney, oniony740745Alcohols2.95 ± 1.42c4.68 ± 2.12a1.09 ± 0.68e3.51 ± 1.91b2.32 ± 0.98d0.93 ± 0.44ef4.49 ± 1.98ab2.72 ± 1.28c0.68 ± 0.34f3.17 ± 1.86c3.88 ± 1.72b0.40 ± 0.21g
Ethyl butanoatefruity805794Esters0.29 ± 0.17b0.05 ± 0.03e0.18 ± 0.09c0.16 ± 0.07c0.10 ± 0.06de 0.21 ± 0.11bc0.44 ± 0.28a0.00f0.00f0.05 ± 0.03e0.00f0.06 ± 0.04e
Butyl acetateappley812812Esters1.27 ± 0.48bc0.00g0.22 ± 0.14e1.71 ± 0.88ab0.05 ± 0.03f0.48 ± 0.32d1.89 ± 0.92a0.00g0.00g1.51 ± 0.74b0.00g0.19 ± 0.08e
(E)-hex-2-enalleafy849847Aldehydes0.16 ± 0.09d0.00e0.73 ± 0.31b0.36 ± 0.14c0.91 ± 0.68a0.96 ± 0.56a0.00e0.44 ± 0.23c0.00e0.67 ± 0.28b0.00e0.74 ± 0.43b
Hexan-1-olflowery865863Alcohols0.6 ± 0.29e1.15 ± 0.72d2.31 ± 0.84c0.79 ± 0.44e0.97 ± 0.63de2.13 ± 1.01c0.62 ± 0.38e9.86 ± 2.92a4.27 ± 1.34b3.79 ± 1.45b10.59 ± 3.18a10.57 ± 3.24a
2-methylbutyl acetatebanana883883Esters4.44 ± 2.72a0.07 ± 0.04f0.00g4.76 ± 2.56a0.00g0.13 ± 0.101.70 ± 0.92c1.09 ± 0.63d1.86 ± 1.32c3.32 ± 1.78b0.63 ± 0.34e0.46 ± 0.22e
Ethyl pentanoatesweet903901Esters0.02 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.02 ± 0.01a0.01 ± 0.01a0.00 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a
Butyl propanoateearthy916915Esters0.01 ± 0.01b0.01 ± 0.01b0.01 ± 0.01b0.02 ± 0.01b0.01 ± 0.01b0.04 ± 0.02a0.01 ± 0.01b0.01 ± 0.01b0.01 ± 0.01b0.01± 0.01b0.01 ± 0.01b0.01 ± 0.01b
Pentyl acetateetheric, fruity, banana923920Esters0.06 ± 0.04a0.01 ± 0.01b0.01 ± 0.01b0.08± 0.06a0.00b0.02± 0.01b0.01± 0.01b0.01± 0.01b0.00b0.00b0.00b0.01± 0.01b
Tert-butyl butanoatefruity, appley955956Esters0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a0.01 ± 0.01a
Hexanoic acidsour, cheesy996996Acids0.41 ± 0.28b0.00c0.00c0.30 ± 0.19b0.00c0.33 ± 0.22b0.00c0.00c0.00c0.00c0.88 ± 0.52a0.00c
Ethyl hexanoatefruity, pineapple1000999Esters9.47 ± 3.12b7.09 ± 2.81de8.18 ± 2.81c7.94 ± 2.63c9.66 ± 2.92ab10.4 ± 3.81a7.78 ± 2.12cd4.75 ± 1.82f4.41 ± 1.76f3.24 ± 1.41g5.13 ± 2.13f6.47 ± 1.96e
Hexyl acetatesweet, green apple, banana10221022Esters0.55 ± 0.28c0.05 ± 0.03f0.20 ± 0.09d0.97 ± 0.41a0.19 ± 0.06d0.89 ± 0.44a0.00g0.10 ± 0.06e0.23 ± 0.11d0.14 ± 0.07de0.70 ± 0.29bc0.81 ± 0.37ab
3-methylbutyl butanoateapple skin10671070Esters0.07 ± 0.04cd0.03 ± 0.02d0.13 ± 0.06bc0.07 ± 0.04cd0.57 ± 0.31b0.19 ± 0.08c0.00e0.17 ± 0.08b0.12 ± 0.04c0.00e0.70 ± 0.49a0.06 ± 0.03d
Ethyl heptanoatefruity, pineapple10981082Esters0.11 ± 0.06cd0.09 ± 0.04d0.13 ± 0.06c0.12 ± 0.05c0.06 ± 0.03e0.18 ± 0.11b0.00f0.07 ± 0.05de0.10 ± 0.04d0.37 ± 0.18a0.14 ± 0.06bc0.07 ± 0.04de
Phenylethyl alcoholflowery, bread-like11161114Alcohols13.92 ± 3.61b7.75 ± 1.04f5.56 ± 0.97g12.34 ± 2.92c8.41 ± 2.42ef7.36 ± 1.52f18.57 ± 3.84a10.40 ± 2.88d9.23 ± 1.82e19.79 ± 4.19a13.14 ± 4.01bc10.18 ± 2.12d
Octanoic acidoily, waxy11901190Acids6.18 ± 3.82a3.76 ± 1.84c1.95 ± 0.74f2.96 ± 1.34de2.96 ± 1.27de 2.43 ± 0.92e5.09 ± 2.24b3.82 ± 1.95c2.09 ± 0.81ef3.09 ± 1.18cd2.11 ± 0.76ef2.45 ± 0.88e
Ethyl octanoatewaxy, musty11971194Esters20.00 ± 2.88de20.68 ± 2.97d23.71 ± 4.84c26.03 ± 5.72bc28.09 ± 5.55b33.02 ± 6.42a21.2 ± 4.14d16.3 ± 2.06f18.66 ± 2.47e14.12 ± 1.62g20.91 ± 3.78d25.97 ± 4.92bc
Hexyl pentanoatefruity, appley, tropical, pear12371235Esters0.05 ± 0.04e0.14 ± 0.08d0.78 ± 0.32b0.00f0.15 ± 0.08d0.92 ± 0.49a0.00f0.06 ± 0.04e0.48 ± 0.31c0.00f0.08 ± 0.06e0.40 ± 0.26c
3-methylobutylhexanoatefruity, waxy, leafy12521253Esters0.07 ± 0.04e0.37 ± 0.19c 0.51 ± 0.28ab0.07 ± 0.05e0.26 ± 0.12d0.44 ± 0.26bc0.00f0.32 ± 0.18c0.57 ± 0.31a0.20 ± 0.11d0.24 ± 0.16d0.27 ± 0.18d
Phenylethyl acetateflowery, honey-like12571255Esters0.42 ± 0.15f0.70 ± 0.31ef0.41 ± 0.17f0.75 ± 0.44ef0.48 ± 0.28f0.33 ± 0.18f0.95 ± 0.62e7.78 ± 4.13a2.91 ± 1.07d4.28 ± 1.41c5.11 ± 1.63b7.87 ± 3.85a
Decanoic acidrotting citrus13771373Acids2.35 ± 1.14bc0.00f1.04 ± 0.38d0.00f1.61 ± 0.78c0.42 ± 0.28e2.93 ± 1.15a2.44 ± 1.06b1.71 ± 0.91c1.82 ± 0.93c1.16 ± 0.56d1.16 ± 0.49d
Ethyl (E)-dec-4-enoateoleic, fruity, pineapple13871379Esters0.00d0.00d0.00d0.36 ± 0.17b0.52 ± 0.24a0.24 ± 0.14c0.00d0.00d0.00d0.32 ± 0.18bc0.24 ± 0.16c0.44 ± 0.21ab
Ethyl decanoateappley, waxy, fruity13951393Esters14.89 ± 2.85f32.10 ± 6.08a28.55 ± 5.91b20.03 ± 4.83de24.47 ± 4.06c21.48 ± 5.34d18.62 ± 3.84e25.62 ± 4.84c33.92 ± 6.41a25.23 ± 4.11c18.93 ± 3.91e18.06 ± 4.02e
Pentyl octanoateirisy, elderberry,14471450Esters0.64 ± 0.28d2.13 ± 0.92b2.87 ± 1.15a0.41 ± 0.15e0.77 ± 0.31d1.05 ± 0.64c0.16 ± 0.09f0.90 ± 0.49cd2.19 ± 0.77b0.48 ± 0.18e0.36 ± 0.11e0.36 ± 0.12e
3-methylbutyl octanoatefruity, waxy14491452Esters0.16 ± 0.09ab0.13 ± 0.07b0.20 ± 0.08a0.00d0.00d0.10 ± 0.04bc0.11 ± 0.04b0.11 ± 0.05b0.00d0.17 ± 0.06a0.11 ± 0.05b0.07 ± 0.04c
Ethyl (2E,4Z)-deca,-4-dienoatefruity, appley, pear14791474Esters0.03 ± 0.02d0.00e0.00e0.19 ± 0.09c0.31 ± 0.19b0.13 ± 0.07c0.00e0.00e0.00e0.50 ± 0.28a0.24 ± 0.16bc0.24 ± 0.15bc
(Z,E)-α-Farnesenewoody14911495Sesquiterpenes0.05 ± 0.04d0.09 ± 0.04bc0.12 ± 0.05ab0.02 ± 0.01e0.08 ± 0.04c0.06 ± 0.03cd0.02 ± 0.01e0.05 ± 0.03d0.13 ± 0.06a0.05 ± 0.04d0.14 ± 0.08a0.06 ± 0.04cd
(E,E)-α-Farnesenewoody, citrusy15071505Sesquiterpenes0.37 ± 0.18cd0.17 ± 0.09f0.35 ± 0.18d0.43 ± 0.22c0.30 ± 0.13de0.18 ± 0.10f0.51 ± 0.31bc0.78 ± 0.34a0.32 ± 0.19d0.26 ± 0.11e0.19 ± 0.08f0.19 ± 0.11f
Dodecanoic acidoily15751570Acids0.24 ± 0.16bc0.51 ± 0.24a0.17 ± 0.10cd0.10 ± 0.04de0.13 ± 0.06d0.10 ± 0.06d0.27 ± 0.13b0.10 ± 0.05de 0.18 ± 0.09c0.00f0.00f0.06 ± 0.04e
Ethyl dodecanoatewaxy, appley, fruity15951595Esters13.68 ± 3.68a14.02 ± 4.15a13.47 ± 3.41a6.61 ± 1.98f6.61 ± 2.12f6.74 ± 2.55ef9.24 ± 2.81c7.40 ± 2.63e11.74 ± 2.88b8.44 ± 2.35d5.26 ± 1.54g4.52 ± 1.01h
1 [17,18,19,20,21,22,23]; 2 I15—Idared apple variety cider fermented at 15 °C; IK15—Idared apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; C15—Cortland apple variety cider fermented at 15 °C; CK15—Cortland apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; G15—Gala apple variety cider fermented at 15 °C; GK15—Gala apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; I20—Idared apple variety cider fermented at 20 °C; IK20—Idared apple (80%) and Konferencja pear (20%) cider fermented at 20 °C; C20—Cortland apple variety cider fermented at 15 °C; CK20—Cortland apple (80%) and Konferencja pear (20%) cider fermented at 20 °C; G20—Gala apple variety cider fermented at 20 °C; GK20—Gala apple (80%) and Konferencja pear (20%) varieties cider fermented at 20 °C. Values are expressed as means (n = 2) ± standard deviations. Mean values with different letters (a, b, c, d, e, f, g, h) within the same column are statistically different (p-value < 0.05).
Table 3. Concentrations of carbohydrates in juices.
Table 3. Concentrations of carbohydrates in juices.
Type of Beverage 1Carbohydrates (w/w)
I10.90 ± 0.15 c
IK11.70 ± 0.15 b
C11.70 ± 0.15 b
CK11.90 ± 0.20 b
G13.70 ± 0.12 a
GK13.90 ± 0.25 a
1 I—Idared apple variety juice; IK—Idared apple (80%) and Konferencja pear (20%) juices mix; C—Cortland apple variety juice; CK—Cortland apple (80%) and Konferencja pear (20%) juices mix; G—Gala apple variety juice; GK—Idared apple (80%) and Konferencja pear (20%) juices mix. Values are expressed as mean (n = 3) ± standard deviation. Mean values with different letters (a, b, c,) within the same column are statistically different (p-value < 0.05).
Table 4. Concentrations of carbohydrates and ethanol and degree of attenuation in ciders.
Table 4. Concentrations of carbohydrates and ethanol and degree of attenuation in ciders.
Type of Beverage 1Ethanol (v/v)Carbohydrates (w/w)Degree of Attenuation (%)
I155.19 ± 0.06e3.95 ± 0.05b63.76f
IK154.88 ± 0.13f3.15 ± 0.07def73.08d
C154.12 ± 0.09h4.38 ± 0.06a60.54g
CK155.18 ± 0.11e2.97 ± 0.04ef75.04c
G156.38 ± 0.08c3.90 ± 0.09b71.53e
GK156.36 ± 0.08c3.82 ± 0.08bc72.52d
I204.92 ± 0.12f2.23 ± 0.05g86.70a
IK205.66 ± 0.20d2.92 ± 0.10ef75.04c
C204.57 ± 0.05g3.08 ± 0.07ef72.25de
CK205.61 ± 0.09d2.80 ± 0.06f76.47b
G206.56 ± 0.14b3.24 ± 0.07de76.35b
GK206.79 ± 0.04a3.49 ± 0.04cd74.89c
1 I15—Idared apple variety cider fermented at 15 °C; IK15—Idared apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; C15—Cortland apple variety cider fermented at 15 °C; CK15—Cortland apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; G15—Gala apple variety cider fermented at 15 °C; GK15—Gala apple (80%) and Konferencja pear (20%) cider fermented at 15 °C; I20—Idared apple variety cider fermented at 20 °C; IK20—Idared apple (80%) and Konferencja pear (20%) varieties cider fermented at 20 °C; C20—Cortland apple variety cider fermented at 15 °C; CK20 Cortland apple (80%) and Konferencja pear (20%) cider fermented at 20 °C; G20—Gala apple variety cider fermented at 20 °C; GK20—Gala apple (80%) and Konferencja pear (20%) varieties cider fermented at 20 °C. Values are expressed as mean (n = 3) ± standard deviation. Mean values with different letters (a, b, c, d, e, f, g) within the same column are statistically different (p-value < 0.05).

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Kliks, J.; Kawa-Rygielska, J.; Gasiński, A.; Głowacki, A.; Szumny, A. Analysis of Volatile Compounds and Sugar Content in Three Polish Regional Ciders with Pear Addition. Molecules 2020, 25, 3564. https://doi.org/10.3390/molecules25163564

AMA Style

Kliks J, Kawa-Rygielska J, Gasiński A, Głowacki A, Szumny A. Analysis of Volatile Compounds and Sugar Content in Three Polish Regional Ciders with Pear Addition. Molecules. 2020; 25(16):3564. https://doi.org/10.3390/molecules25163564

Chicago/Turabian Style

Kliks, Jarosław, Joanna Kawa-Rygielska, Alan Gasiński, Adam Głowacki, and Antoni Szumny. 2020. "Analysis of Volatile Compounds and Sugar Content in Three Polish Regional Ciders with Pear Addition" Molecules 25, no. 16: 3564. https://doi.org/10.3390/molecules25163564

APA Style

Kliks, J., Kawa-Rygielska, J., Gasiński, A., Głowacki, A., & Szumny, A. (2020). Analysis of Volatile Compounds and Sugar Content in Three Polish Regional Ciders with Pear Addition. Molecules, 25(16), 3564. https://doi.org/10.3390/molecules25163564

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