Determination of the physicochemical profile and volatile compounds of meads produced in different regions of Brazil

Abstract

Mead, a traditional alcoholic beverage produced through the fermentation of diluted honey by yeasts, remains underexplored in terms of its quality and characteristics. To establish quality standards for this artisanal beverage, it is essential to evaluate the physicochemical properties and sensory attributes of different mead types. This study aimed to investigate the physicochemical properties and volatile organic compound (VOC) profiles of meads produced in diverse regions of Brazil. Mead samples with varying production times and characteristics, such as smooth, aged with French oak chips, special reserve, were analyzed. The parameters examined included ash content, pH, total acidity, dry extract, total soluble solids, and VOCs. The VOC profile was determined using the HS-SPME-GC-MS technique. The physicochemical parameters were determined as follows: pH (2.80–3.60), total acidity (47.33–115.33 meq L⁻¹), ash content (0.59–1.47 %), dry extract (35.12–104.60 g L⁻¹), and total soluble solids (6.00–13.73 °Brix). A total of 28 VOCs were identified, classified into volatile acids (5), alcohols (3), esters (15), and terpenes (5). The variation in physicochemical properties and VOC profiles was primarily influenced by the raw materials used and the aging period of the meads. This study provides valuable insights into mead quality, offering a comprehensive characterization of its physicochemical and aromatic attributes, and contributes to the establishment of quality standards for mead production.

Key words
Alcoholic beverage; Apis mellifera L; craft drink; honey; multivariate analysis

INTRODUCTION

Mead is one of the oldest beverages consumed by humans, predating wine and often considered a precursor to beer. It contains 4 to 18% (v/v) alcohol and is produced through the alcoholic fermentation of a mixture of honey, nutrient salts, and potable water. In mead production, fermentation is a crucial process, and nutrient salts such as di-ammonium phosphate (DAP) or mixed supplements play an important role in ensuring that the yeast has the necessary resources to ferment the sugars in honey and water, converting them into alcohol and carbon dioxide (Brazil 2009, Kružík et al. 2022, Webster et al. 2025).

Several studies suggest that moderate consumption of mead may help prevent various health issues such as diabetes, cancer, inflammation, respiratory infections, and gastrointestinal disorders, attributed to the presence of organic acids, phenolics, and peptides in honey (Essiedu & Kovaleva 2024). Consequently, the global mead market is projected to expand at a compound annual growth rate of 18.71%, increasing from USD 432.4 million in 2020 to USD 1,621.0 million by 2028 (Fortunebusinessinsight 2024).

While most honey is traditionally used in complementary medicine, the brewing industry has incorporated it into the production of alcoholic beverages (Starowicz & Granvogl 2020). Mead can be handcrafted and enhanced with additives such as citric acid, herbs, spices, fruit pulp, or juice (Mascarenhas et al. 2017). It possesses a light hue, akin to honey, with a delicate, floral aroma and a taste that strikes a balance between sweet and dry (Iglesias et al. 2014).

In Brazil, mead remains a relatively under-consumed beverage, but it holds significant market potential, especially given the growing trend of craft beer consumption in the country (Nakada et al. 2020). Although a few commercial meaderies have already been established, the majority of production remains informal, and there are currently no official statistical data on the production and consumption of mead in Brazil (Brunelli et al. 2017).

It is essential to emphasize that honey quality parameters, such as moisture content, free acidity, pH, ash content, sugar composition, soluble solids concentration, color, and mineral content, significantly influence the composition of meads (Nordin et al. 2018). Different types of meads may exhibit distinct physicochemical properties; however, these characteristics must comply with the standards established by Brazilian legislation, specifically Normative Instruction No. 34 (Brazil 2012), which sets guidelines for all fermented beverages, including mead.

Volatile compounds play a crucial role in defining the sensory quality of beverages. They directly affect the perception of flavor, aroma, and freshness, as well as influencing consumer acceptance. Controlling these compounds is essential for producing high-quality beverages and ensuring market differentiation (Feng et al. 2024).

Therefore, the formation of volatile organic compounds (VOCs) during mead fermentation results from a complex interaction between honey sugars, yeast activity, and fermentation conditions. These VOCs, including alcohols, esters, acids, and aldehydes, play a crucial role in shaping the aromatic profile of mead. The selection of ingredients, yeast strains, and fermentation parameters directly influences the quality and diversity of the final beverage’s aroma and flavor (Starowicz & Granvogl 2022, Webster et al. 2025). This study aimed to assess the physicochemical profile and volatile organic compound (VOC) composition of meads produced in different regions of Brazil, considering variations in production time and aging processes.

MATERIALS AND METHODS

Standards and reagents

Sodium hydroxide (PA grade, NEON), standard solutions of n-alkanes (C₇–C₃₀), ethyl hexanoate, ethyl octanoate, decanoic acid, dodecanoic acid, tetradecanoic acid, ethylhexyl, oleic acid, hexadecanoic acid, 1-octadecanol were acquired from Sigma–Aldrich (USA). All standards used in GC–MS analysis had a purity of ≥ 95%. The SPME fiber (Divinylbenzene/Carboxen/Poly(dimethylsiloxane) (DVB/CAR/PDMS, 65 μm) and the SPME holder for manual sampling were purchased from Supelco (Bellefonte, PA, USA). All solutions were prepared using distilled water obtained from a reverse osmosis system (LT 800/20, Limatec, Brazil).

Obtaining the mead

Table I presents the characteristics of mead samples obtained from different regions of Brazil, labeled 1 to 6 to maintain brand confidentiality. Samples 1 and 4 were commercially produced and registered, while samples 2, 3, 5, and 6 were artisanal products. Mead samples 3 and 6 originated from the same location and production batch; however, sample 6 was analyzed immediately after production, whereas sample 3 was analyzed six months post-production. The alcohol contents presented in Table I were obtained directly from the label of each product.

Table I
Characteristics of mead samples obtained from different regions of Brazil.

Nº Mead	Origin	Alcohol Content (% v/v)	Description
1	São Paulo (23°33’S 46°38”W)	13.0	Smooth
2	Recôncavo of Bahia (12°40’29.9”S 39°06’9.9”W)	7.0	Smooth
3	Recôncavo of Bahia (12°40’29.9”S 39°06’9.9”W)	8.0	Smooth aged in French oak chips
4	Diamantina Plateau (-12° 52’ 49.44” S -41° 22’ 20.28” W)	14.0	Special reserve
5	Recôncavo of Bahia (13º 02’ 06” S 39° 00’ 52” W)	7.0	Smooth
6	Recôncavo of Bahia (13º 02’ 06” S 39° 00’ 52” W)	8.0	Smooth aged in French oak chips

Physical-chemical characterization

The pH was measured using the potentiometric method with a digital pH meter (Q-400A, Quimis). Total titratable acidity (meq L^-1), based primarily on the measurement of tartaric, malic, acetic and lactic acids, was determined by titration with 0.1 M sodium hydroxide solution. Ash content was quantified using the gravimetric method, with samples incinerated at 550 ± 25°C in a muffle furnace. The dry extract was obtained by weighing the residue after evaporation in a water bath and drying in an oven at 100 ± 5°C, using the formula: (1000 × dry residue mass) / sample volume. Total soluble solids (TSS) were measured with a digital benchtop refractometer (Kruss, Hamburg, Germany) (Instituto Adolfo Lutz 2008). All analyses were conducted in triplicate.

Extraction of volatile organic compounds

VOCs were extracted in triplicate using the headspace solid-phase microextraction (HS-SPME) technique (Rodrigues et al. 2011). Ten milliliters of each mead sample was placed in a 20 mL vial under agitation at 171 rpm. A 20 minute equilibrium period was established to allow VOCs to concentrate in the headspace. The vial was then placed on an aluminum block on a hot plate set at 85°C. Using a sampler, the vial’s seal was pierced, and a 65 μm Supelco® DVB/CAR/PDMS fiber was exposed for 78 min to absorb volatile compounds (Nascimento et al. 2024). Subsequently, the fiber was removed from the vial and inserted into the gas chromatograph injector for thermal desorption of the analytes at 250°C for 3 minutes.

Analysis of volatile organic compounds by GC-MS

The extracted compounds were separated by gas chromatography-mass spectrometry (GC-MS), model GCMS-QP2010 Plus (Shimadzu, Kyoto, Japan), with an electron impact ionization source at 70 eV. The desorption/injection of the analytes into the column was performed with an injector in the flow split mode (15:1). The chromatographic separation was performed on an HP–5 MS column ([5%-Phenyl]-methylpolysiloxane; 30 m × 0.25 mm ID × 0.25 μm), with the following oven heating program: initial temperature of 40 °C for 5 min, followed by a ramp to 40 °C (0.5 °C min-1) for 2 min, then to 60 °C (1 °C min-1), then to 100 °C (1 °C min-1) for 5 min, then to 150 °C (2 °C min-1) for 5 min, then to 200 °C (2 °C min-1) for 5 min, then to 250 °C (4 °C min-1), and finally up to 300 °C (15 °C min-1), totaling a run of 166 min. The helium carrier gas was maintained at a constant flow rate of 0.60 mL min-1 (12.8 Kpa) in the column, with a constant linear velocity of 27.9 cm s-1. The transfer line and ionization source temperatures were 230 °C.

Identification and analysis of volatile organic compounds

VOCs were identified by calculating linear retention indices, comparing mass spectra obtained from the analyses with those in the library (NIST 147 Database), and injecting a homologous series of n-alkanes (C₇ to C₃₀). To confirm compound identification, their peak areas were compared with those of analytical standards purchased from Sigma-Aldrich®.

Statistical analysis

The results of the physicochemical analyses were expressed as mean ± standard deviation. Analysis of variance (ANOVA), followed by Tukey’s test, was performed to determine statistically significant differences between means (p < 0.05) using XLStat software (version 7.8). The relative areas of the detected and identified compounds were autoscaled and subjected to multivariate principal component analysis (PCA) and hierarchical cluster analysis (HCA) using the Metaboanalyst 5.0 software to assess group formation among the different samples based on the similarity and relative values ​​of VOCs areas (Mesquita et al. 2017).

RESULTS AND DISCUSSION

Physicochemical characterization

As a result of the physicochemical analyses, the data presented in Table II outline the parameters for pH, total titratable acidity, ash content, dry extract, and total soluble solids. Given the varying alcohol content, compositions, and shelf life of the meads used in this study, these factors appear to directly influence their physicochemical properties. In this context, considering that mead is an emerging sector within the alcoholic beverage industry in Brazil, and is expanding rapidly in the United States, different compositions, diverse ingredients, and varying preparation methods and shelf lives have been explored to produce a high-quality beverage with strong consumer acceptability, despite potential alterations in its physicochemical characteristics (Nakada et al. 2020).

Table II
Physicochemical analysis of meads produced in different regions of Brazil.

Parameters evaluated	H1	H2	H3	H4	H5	H6
pH	3.49±0.02a	2.80±0.01e	2.84±0.03e	3.60±0.03b	3.13±0.02c	2.90±0.01d
Total acidity (meq L-1)	47.33±0.23e	58.93±0.83b	59.33±1.15b	115.33±1.15a	54.27±0.46c	50.13±0.23d
Ash (%)	1.47±0.01a	0.97±0.01ab	1.11±0.03ab	1.20±0.06ab	0.59±0.03b	0.65±0.03b
Dry extract (g L-1)	91.86±3.71b	97.65±0.09ab	44.95±0.11d	104.60±1.80a	65.08±4.59c	35.12±1.38e
Total soluble solids (°Brix)	11.00±0.00a	10.20±0.00b	6.00±0.00fc	13.73±0.29d	8.87±0.23e	6.87±0.12f

• Averages followed by the same letters, in the line, do not differ from each other at 5%, according to the Tukey test.

Regarding the pH of the analyzed formulations, statistically significant differences were observed. However, all values were ≤ 4.00, classifying them as acidic, a similar result was reported by Oliveira et al. (2020) in a study on sweet mead. In the present study, a comparison between the pH of mead produced six months before analysis (H3) and the mead produced in the month of analysis (H6) revealed a statistically significant difference, with H3 being more acidic than H6 mead. This suggests that the shelf life may have influenced these values, indicating that this type of mead tends to become more acidic over time. Additionally, the meads were aged with oak chips, which, besides imparting a woody flavor to the beverage, may also affect its physicochemical parameters throughout the aging process.

According to Kawa-Rygielska et al. (2019), a significant decrease in pH can impair yeast performance during fermentation. Therefore, it is crucial to monitor pH throughout mead production, particularly during fermentation, to ensure that the environment remains conducive to yeast development, which requires both technical and scientific expertise. Additionally, proper management of the finished product is essential, as physicochemical changes over time may lead to the development of undesirable aromas and flavors, especially in the presence of inactive yeasts, which can negatively impact the beverage’s acceptability (Akalin et al. 2017).

Acidity plays a fundamental role in alcoholic beverages, directly influencing their flavor and stability. Brazilian legislation mandates that meads have a total titratable acidity between 50 and 130 meq L^-1 (Kawa-Rygielska et al. 2019). In this study, significant differences were observed among the formulations for this parameter, with values ranging from 47.33 to 115.33 meq L^-1. Notably, formulation H1 had a value below the recommended minimum limit. Musachio et al. (2022) reported values of 59.2 and 57.35 meq L^-1 for standard and flavored mead, respectively, both of which fall within the legislated range.

In this context, pH and acidity play crucial roles in various stages of the mead fermentation process. They influence fermentation kinetics, the production of volatile organic compounds, microbiological stability, and the sensory quality of the beverage. Proper control of pH and acidity is essential for optimizing the production of desirable aromas, ensuring the stability of the final product, and maintaining the quality and safety of mead for consumers. Additionally, balancing these factors is important to prevent excessive acidity, which could impair palatability or pose potential health risks (Essiedu et al. 2021, Kawa-Rygielska et al. 2019, Webster et al. 2025).

However, H4 exhibited the highest acidity (115.33 meq L^-1) and the highest pH value (3.60). Typically, during fermentation, pH decreases as total acidity increases due to the formation of organic acids by yeasts. Nevertheless, abnormal variations in total acidity and pH may indicate the presence of contaminants (Almeida et al. 2017). In this regard, Tôrres et al. (2011) emphasized that the physicochemical parameters of mead are closely related to fermentation performance, sensory characteristics, and product stability, with acidity being a key attribute that determines the beverage’s quality.

The ash content of the analyzed meads ranged from 0.59% to 1.47% across the formulations, which complies with NI 34 (Brazil 2012), which stipulates a minimum ash content of 0.15%. Musachio et al. (2022) assessed the physicochemical characteristics of both standard mead and mead flavored with red grape pomace, comparing their results to NI 34 (Brazil 2012). They found ash contents of 1.85% and 2.29%, respectively, which are also consistent with the requirements of the Normative Instruction. This suggests that the inclusion of grape pomace in the formulation may have contributed to the increased ash content.

Total dry extract represents the weight of all non-volatile substances when exposed to specific physical conditions, including fixed acids, organic and mineral salts, polyalcohols, phenolic compounds, nitrogen compounds, sugars, and polysaccharides (Rizzon 2010). Beverages with a total dry extract exceeding 20.00 g L^-1 are classified as full-bodied, while those with lower values are considered light (Aquarone et al. 2001). In this study, the total dry extract of the final mead ranged from 35.12 to 104.60 g L^-1, categorizing the analyzed meads as full-bodied. Although the table specifying the standard of identity and quality for meads does not set limits for total dry extract, it does specify limits for reduced dry extract, which is the total dry extract minus excess total sugars (Rizzon 2010). The legislation mandates a minimum of 7.00 g L^-1 for this attribute (Brazil 2022).

Another attribute closely related to beverage quality is the total soluble solids (TSS), which significantly contribute to flavor. The mead samples exhibited TSS values ranging from 6.00 to 13.73 °Brix, with the H3 and H6 meads showing the lowest levels. These formulations were similarly aged with French oak chips. A study on wine production using French oak chips found that they enhanced microbiological stability, significantly affected color parameters, and increased the total polyphenol content (Silva et al. 2020).

Lopes et al. (2020) investigated a fermented alcoholic beverage made from cajarana and reported a TSS content of 10.00 °Brix, which is comparable to the values observed in this study. They also noted a decrease in °Brix during fermentation, from 23.30 °Brix on the first day to 10.00 °Brix on the eighth day. However, the highest TSS content was found in sample H4, suggesting that higher soluble solids content may correlate with higher acidity. This result implies potential inhibition of yeasts during fermentation, leading to a reduced use of soluble solids for acid production. Additionally, a correlation exists between TSS and mead acidity, as a higher content of soluble solids can lead to increased production of organic acids during fermentation, thereby raising acidity (Abrol & Joshi 2012).

Table II reveals a clear correlation between total soluble solids (°Brix) and dry extract for the evaluated mead samples, indicating that higher content of soluble solids is associated with higher dry extract content. A similar correlation was observed by Almeida et al. (2017) in brandy samples made from Ziziphus joazeiro bark. This is because a higher concentration of sugars in the must (measured in °Brix) leads to a greater amount of solid matter, including unfermented sugars and minerals, remaining after evaporation, which consequently results in a higher dry extract.

Determination of volatile compounds in mead

A total of 28 volatile organic compounds (VOCs) were identified in the mead samples, including 5 volatile acids, 3 alcohols, 15 esters, and 5 terpenes, with nine of these compounds being confirmed using analytical standards (Table III). Fig. 1 presents a representative chromatogram of the VOCs analysis of the mead samples by GC-MS.

Figure 1
Representative chromatogram obtained from the GC-MS analysis of a mead sample.

Table III
Average relative composition (%) of VOCs identified in the different mead samples.

No.	Compound	LRIbEXP	LRIcLIT	MS Similarity (%)	Mead Samples
					H1	H2	H3	H4	H5	H6
1	2,3-butanediol	809	806	96	1.85±1.02a	0.73±0.68a	5.54±1.74b	2.11±1.44ab	0.90±0.57ab	-
2	isopentyl acetate	879	879	94	0.45±0.03a	0.24±0.04b	0.46±0.06a	2.91±1.02c	0.61±0.01a	0.37±0.02a
3	ethyl hexanoatea	1003	1003	94	1.85±0.43a	1.15±0.12b	4.06±0.24c	1.90±1.72a	1.68±0.06a	1.02±0.71b
4	(E)-linalool oxide	1070	1070	91	1.23±0.39a	4.31±0.20b	10.07±1.55c	3.28±0.73b	4.03±0.29b	4.67±0.49b
5	(Z)-linalool oxide	1080	1081	89	-	0.52±0.03a	-	-	0.33±0.08a	-
6	hotrienol	1108	1108	94	5.76±0.34a	5.14±2.12a	3.79±0.09b	37.53±8.58c	12.69±3.14a	7.17±3.05a
7	phenethyl alcohol	1109	1109	83	-	0.51±0.60a	-	-	-	-
8	(E,E)-cosmene	1131	1130	93	-	13.76±0.76a	2.29±0.17b	0.53±0.43c	12.84±1.41a	2.11±0.62b
9	nerol oxide	1157	1155	91	-	9.96±0.30a	3.30±0.35b	0.50±0.46c	7.57±0.84a	1.08±0.17c
10	diethyl succinate	1189	1188	94	3.72±0.62a	0.41±0.04b	2.91±0.72a	9.53±2.98c	1.55±0.19d	1.46±0.13d
11	ethyl octanoatea	1200	1200	95	11.79±1.17a	5.11±1.72b	15.84±1.63c	2.89±1.49b	8.95±0.49d	11.89±1.06a
12	ethyl benzeneacetate	1245	1245	90	0.40±0.11a	0.12±0.01b	0.35±0.11a	0.43±0.54a	0.40±0.05a	0.20±0.01ab
13	decanoic acida	1386	1386	81	3.23±0.56a	-	3.03±0.87a	-	3.77±0.10a	3.26±0.48a
14	ethyl dec-9-enoate	1390	1390	81	-	4.52±0.68a	0.83±0.23b	-	0.34±0.40b	2.85±0.40c
15	ethyl decanoate	1398	1398	95	7.58±2.02a	1.52±0.98b	2.27±0.64b	0.10±0.24c	2.17±0.32b	6.41±2.40a
16	dodecanoic acida	1571	1571	85	-	0.10±0.04a	-	-	-	0.34±0.06b
17	ethyl dodecanoate	1596	1596	92	0.63±0.24a	0.13±0.06b	0.16±0.02b	-	0.11±0.01b	0.91±0.62c
18	tetradecanoic acida	1768	1768	93	-	0.24±0.13a	0.08±0.04b	-	-	-
19	2-ethylhexyl salicylatea	1799	1805	94	2.89±0.88a	3.51±0.53a	3.94±2.19a	1.36±0.25b	1.24±1.37b	4.32±1.50a
20	isopropyl myristate	1833	1832	84	-	0.50±0.08a	-	-	-	-
21	methyl salicylate	1876	1903	95	2.11±1.39a	0.79±0.68b	-	-	-	-
22	hexadecanoic acida	1965	1965	93	2.96±0.64a	3.70±0.42a	4.33±1.38a	1.29±0.41b	1.99±1.06a	2.80±0.86a
23	ethyl 9-hexadecanoate	1972	1975	83	-	-	-	-	0.28±0.49a	-
24	ethyl hexadecanoate	1995	1995	91	4.37±0.36a	3.13±0.26b	2.25±0.16c	1.07±0.08d	3.01±0.33b	0.88±0.10d
25	1-octadecanola	2081	2081	96	2.41±0.60a	0.53±0.38b	1.00±0.68b	0.27±0.46b	0.52±0.21b	1.85±0.59c
26	oleic acida	2133	2131	85	-	0.12±0.02a	-	-	-	-
27	ethyl oleate	2163	2168	92	0.47±0.24a	1.61±0.19b	2.53±0.14c	0.91±0.34d	4.97±0.52e	1.00±0.08d
28	ethyl octadecanoate	2193	2193	89	0.70±0.40a	1.02±0.07a	1.08±0.15a	0.09±0.21b	2.03±0.22c	0.55±0.34a

• aIdentification confirmed by comparison with mass spectra and retention times of analytical standards. _LRIbEXP = linear retention index relative to the n-alkane stand_ard (_C7 a C30) obtained on a HP–5 MS capillary column. _LRIcLIT =linear retention index published in the literature (NIST Chemistry Web Book, accessed in www.webbook.nist.gov). n = 3 replicates per sample. Means followed by the same letter on the line does not differ significantly from each other (Tukey test; p<0.05). (-): compound not detected.

During fermentation, yeasts convert the sugars in honey into ethanol while simultaneously producing various organic acids, such as acetic acid, lactic acid, and butyric acid. These acids can react with alcohols, including ethanol, to form esters. Controlling ester production during fermentation is essential for balancing the sensory properties of mead. The optimal ester concentration depends on the desired style of mead and the intended sensory profile. Generally, lower ester concentrations contribute to a more pleasant and complex sensory experience, while higher concentrations may result in an unbalanced sensory profile (Chitarrini et al. 2020, Kuś et al. 2022, Starowicz & Granvogl 2020).

Esters represent the largest group of volatiles in meads, with ethyl octanoate and ethyl decanoate being present in higher concentrations, statistically different between the samples analyzed. These esters contribute fruity and cognac-like aromas and were also identified in the study by Pino & Fajardo (2011), which evaluated beverages made with unifloral honeys. According to Fu et al. (2023), the formation of esters during fermentation is influenced by nitrogen levels. Additionally, some esters can be degraded in acidic environments, which may explain the lower levels of esters observed in mead H4, which has a higher total acidity.

Terpene production during the fermentation of beverages such as mead can be influenced by various factors, including the yeast strain and fermentation conditions. The concentration of terpenes significantly impacts the sensory properties of the beverage. Low terpene concentrations contribute to a smoother, more balanced sensory profile, while high terpene concentrations can intensify both aroma and flavor. Modulating terpene levels is crucial for developing beverages with desirable and well-balanced sensory characteristics (Romano et al. 2021, Starowicz & Granvogl 2022). Terpenes constitute the second-largest group of volatiles, with hotrienol being the most prominent, statistically different from samples H3 and H4. It was detected in all samples, with the highest concentrations found in H4 (37.53%) and H5 (12.69%) meads. Hotrienol, which has a floral aroma, was also observed in high concentrations in the study by Pino & Fajardo (2011).

During fermentation, yeasts produce not only ethanol but also a variety of organic acids as byproducts. These acids are generated through secondary metabolic pathways involving the degradation of sugars and amino acids, or by the activity of microorganisms such as lactic acid bacteria. Low concentrations of acids contribute to balance and freshness, while high concentrations may lead to an overly acidic sensation on the palate. Controlling acidity is essential for production beverages with a well-defined structure and sensory harmony (Pereira et al. 2019, Starowicz & Granvogl 2022). Among the acidic compounds, hexadecanoic acid is the most prominent. Fu et al. (2023) similarly found hexadecanoic acid to be present in higher concentrations compared to other acids in mead samples.

The production of alcohols during fermentation involves the conversion of sugars into ethanol. At low concentrations, these alcohols can enhance complexity and subtlety, while at high concentrations, they may impart bitter notes or undesirable characteristics (Pereira et al. 2019, Romano et al. 2021, Webster et al. 2025). Regarding alcohols, 2,3-butanediol was detected in all mead samples except H6, with the highest concentration in H3 (5.54%). The absence of this compound in H6 may be attributed to the time elapsed since production, as samples H3 and H6 originated from the same location and production batch. Sample H6 was analyzed shortly after production, while sample H3 was analyzed six months later. 2,3-butanediol, which has a fruity aroma, has been detected in meads produced with yeasts that have low nitrogen requirements (Schwarz et al. 2020).

It is important to emphasize that many volatile organic compounds (VOCs) in alcoholic beverages are formed during the fermentation process and are primarily influenced by factors such as the content of reducing sugars, water activity, pH, as well as fermentation time and temperature (Jousse et al. 2002). Additionally, the type and origin of honey play significant roles in determining the composition and concentration of VOCs in mead (Starowicz & Granvogl 2020).

Multivariate analysis

The profile of volatile organic compounds (VOCs) identified in the samples was evaluated using multivariate analysis techniques. This approach enabled the grouping of samples based on the similarity of their VOC compositions, as illustrated by the dendrogram in Figure 2. The dendrogram reveals the formation of six distinct clusters, each corresponding to a different mead sample. Mead samples H3, H4, and H5 (dark blue, light blue, and pink, respectively) exhibit similar VOC profiles, grouping together within a larger cluster. In contrast, samples H1 and H6 (red and yellow, respectively) form a separate cluster, suggesting a closer similarity between them. Notably, sample H2 (green) displays a markedly distinct VOC profile, setting it apart from the other samples.

Figure 2
Dendrogram associated with the heat map of VOCs identified in the mead samples used in this study. Above: dendrogram with the types of mead used; on the left side: dendrogram of the identified compounds. The color of the cells refers to the abundance of the compound in the VOC profile. Red represents high abundance and blue low abundance.

The heat map analysis reveals that ethyl hexadecanoate, (Z)-linalool oxide, (E,E)-cosmene, nerol oxide, phenethyl acid, tetradecanoic acid, isopropyl myristate, and oleic acid are more abundant in the green cluster (H2), indicating that this mead is particularly rich in esters and alcohols. Although these compounds are also present in mead H5, their concentrations are lower compared to H2. Notably, both meads share the same alcohol content (7.00%), suggesting that factors beyond ethanol concentration, such as fermentation conditions and raw material composition, may influence the volatile profile.

Mead sample H3 exhibits higher concentrations of (E)-linalool oxide, 2,3-butanediol, and ethyl hexanoate, the latter being a key ester produced produced by yeast during fermentation. In contrast, H4 is distinguished by the presence of isopentyl acetate, hotrienol, diethyl succinate, and ethyl benzeneacetate, contributing to its unique volatile profile. Meanwhile, the volatiles ethyl octanoate, decanoic acid, ethyl decanoate, ethyl dodecanoate, and homomenthyl salicylate are more concentrated in H1 and H6, which differ in both composition and alcohol content, highlighting the influence of formulation and fermentation conditions on volatile compound production.

Medeiros & Niro (2022) compiled a comprehensive table of the key volatile compounds found in meads produced with different ingredients and additives, reporting results closely aligned with the volatiles identified in this study. A diverse range of esters, alcohols, aldehydes, and terpenes were detected, primarily originating from the honey and other raw materials used, as well as from the fermentation process, further highlighting the influence of ingredient composition and fermentation conditions on mead’s volatile profile.

Principal component analysis (PCA) was performed, as illustrated in Figure 3. The score plot (Fig. 3a) reveals cluster formations consistent with those observed in the hierarchical cluster analysis (HCA) dendrogram. The grouping of samples according to mead type further validates the ability of the HS-SPME/GC-MS technique to effectively characterize the VOC profiles of the analyzed meads, highlighting its reliability in differentiating samples based on their volatile composition.

Figure 3
2D PCA score plot for different mead samples (a). Biplot of principal component analysis (PCA) illustrating the distribution of volatile compounds (variables) in the mean of each treatment (samples) (b).

The score plot clearly illustrates a distinct separation in the VOC profiles for each mead type, with H2 samples exhibiting a markedly different profile compared to the other analyzed meads. Figure 3b presents the PCA loadings plot, highlighting the VOCs that most significantly contribute to the differentiation of each sample group, based on the mean values of each treatment.

By overlaying the volatile compounds with the samples in the graph (Fig. 3b), it becomes evident that each mead type is characterized by a distinct set of key VOCs. In H1 samples, decanoic acid, ethyl octanoate, and 1-octadecanol are the predominant volatiles. H2 is distinguished by the presence of isopropyl myristate, phenethyl alcohol, tetradecanoic acid, hexadecanoic acid, and ethyl dec-9-enoate. The volatile profile of H3 is defined by 2,3-butanediol, diethyl succinate, and ethyl hexanoate. H4 is characterized by isopentyl acetate, ethyl benzeneacetate, and hotrienol. In H5, ethyl 9-hexadecanoate, ethyl oleate, and (E)-linalool oxide are the defining compounds. Finally, H6 samples are marked by the presence of ethyl decanoate and ethyl dodecanoate.

In a study by Pino & Fajardo (2011) on the volatile composition of fermented alcoholic beverages made from unifloral honeys, 23 compounds were identified, including alcohols, esters, aldehydes, and terpenes, with esters and terpenes being the most abundant classes. Figure 4 highlights some of the key volatile compounds that contribute to the differentiation of mead samples. Nerol oxide is a monoterpene commonly found in various essential oils. The highest concentration of this compound was observed in H2, followed by H5. Both meads are of the mild type, with an alcohol content of 7.00%. Ethyl oleate, a fatty ester used in the food, cosmetics, and pharmaceutical industries (Dias et al. 2022), was present in higher concentrations in H5 and lower in H6.

Figure 4
Box Plot for some of the most significant VOCs in discriminating mead samples (a) nerol oxide (b) ethyl oleate (c) decanoic acid (d) ethyl decanoate (e) dodecanoic acid (f) ethyl hexadecanoate.

Decanoic acid, also known as capric acid, is a carboxylic acid with the formula CH₃(CH₂)₈COOH, and is found in higher concentrations in meads H6 and H1. This compound has been identified in fermented alcoholic beverages made from artisanal jabuticaba (Dias et al. 2022) and Tannat wine (Wei et al. 2018). Decanoic acid, along with ethyl decanoate, was also detected in a study evaluating the volatile composition and sensory properties of meads produced with free or immobilized Saccharomyces cerevisiae cells (Pereira et al. 2019). Ethyl decanoate was present in all samples, with the lowest concentration found in H4, which exhibited the highest alcohol content. This ester, characterized by a longer carbon chain, has lower volatility, a higher odor threshold, and influences texture; it is commonly found in cachaça (Nóbrega 2003). Ethyl hexadecanoate was present in all mead formulations, with higher concentrations in H2 and H5. This ester is produced during alcoholic fermentation via the secondary intracellular metabolism of yeast (Nascimento et al. 2009).

Identifying the volatile organic compounds (VOCs) present in a beverage is essential for assessment its flavor and aroma quality, key attributes that not only in influencing consumer preference but also help detect potential adulteration. Therefore, determining these compounds is essential for both final product analysis and production process control, as the profile of VOCs can vary depending on the type of beverage and may be formed or concentrated during fermentation, distillation, and maturation stages.

It is important to note that the analysis of variations in the physicochemical profiles and volatile compounds of meads from different geographic origins remains an unexplored approach. Including multiple Brazilian regions enables the identification of patterns and significant variations that can directly influence mead quality, offering novel data that contribute to enhancing production practices and fostering the development of higher-quality products in the market.

CONCLUSIONS

This study demonstrated that the employed approach effectively determined the physicochemical attributes and volatile organic compound profiles of meads from different origins. Both physicochemical characteristics and VOCs profiles varied depending on the raw materials used and the beverage’s shelf life. A total of 28 VOCs were identified, allowing differentiation among the various mead types based on the profiles of esters, alcohols, and carboxylic acids. These findings highlight the potential of VOC profiling as a valuable tool for assessing the quality and unique characteristics of meads from different origins. Additionally, future research should focus on profiling bioactive compounds, total phenolic content, total flavonoid content, and antioxidant activity, as well as assessing consumer preferences and how these factors influence market acceptance. This would aid in the improvement of production techniques and the diversification of mead offerings.

Acknowledgements

This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Process No. 305950/2021-5) and the Maria Milza University Center, which awarded a full Master’s scholarship to Guedes ISA and Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior (CAPES; No. 88887.504355/2020-00) which awarded a full Master’s scholarship to Nascimento MB.

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Publication Dates

• Publication in this collection
  31 Oct 2025
• Date of issue
  2025

History

• Received
  06 Jan 2025
• Accepted
  17 May 2025