Sugarcane variety as a determinant of agro-industrial performance and physicochemical quality in pot still cachaça production

Variedade de cana-de-açúcar como determinante do desempenho agroindustrial e da qualidade físico-química na produção de cachaça de alambique

ABSTRACT

The predominance of a single sugarcane variety (RB867515) or unidentified genotypes in cachaça production may limit both agronomic efficiency and beverage quality. This study evaluated four alternative sugarcane varieties (RB966928, RB036066, CTC9002, and CTC9003) for their impact on agronomic performance and the physicochemical properties of pot still cachaça. Field trials were conducted at Cachaçaria João Mendes Distillery in Perdões, MG, Brazil, with milling, fermentation, and distillation carried out at Cachaçaria João Cassiano in Candeias, MG, Brazil, during the 2022/23 harvest season. Key agronomic and industrial parameters, such as total soluble solids (TSS, °Brix), maturity index (MI), expected cachaça yield (ECY), tons of stalks per hectare (TSH), and liters of cachaça per ton (LCT), varied significantly among the varieties. The varieties CTC9002 and RB036066 exhibited the highest TSH, while CTC9003 exhibited the highest concentration of total soluble solids in the must (TSSM, ºBrix). The pH values across all varieties ranged from 4.00 to 4.33. The average yeast and bacterial counts were 10⁷ and 10³ CFU/mL, respectively. All pot still cachaça samples, regardless of sugarcane variety, complied with the physicochemical standards established by Brazilian legislation. Overall, the varieties CTC9002 and RB036066 demonstrated strong potential for diversifying sugarcane cultivation in cachaça-producing regions.

Index terms:
Distilled beverage; production system; fermentation; Saccharum spp.; quality beverage

RESUMO

Nas agroindústrias de cachaça, predominam o cultivo único da variedade RB867515 e o de variedades desconhecidas de cana-de-açúcar que podem afetar o rendimento agronômico e a composição da bebida. Portanto, o objetivo deste estudo foi avaliar novas variedades de cana-de-açúcar (RB966928, RB036066, CTC9002 e CTC9003) e seus efeitos no rendimento agronômico e na composição físico-química da cachaça de alambique. O experimento de campo foi conduzido na Cachaçaria João Mendes (destilaria), no município de Perdões, MG, Brasil. A moagem, a fermentação e a destilação da cana-de-açúcar foram conduzidas na Cachaçaria João Cassiano, Candeias, MG, Brasil, na safra 2022/23. Foram avaliados os caracteres agronômicos da cana-de-açúcar, o rendimento e os componentes físico-químicos da cachaça. As variedades apresentaram diferenças significativas para os seguintes parâmetros: sólidos solúveis totais (SST, °Brix), índice de maturação (IM), rendimento esperado de cachaça (REC), toneladas de colmos por hectare (TCH) e litros de cachaça por tonelada (LCT). As variedades CTC9002 e RB036066 apresentaram os melhores resultados em toneladas de colmos por hectare (TCH). No momento da fermentação, os sólidos solúveis totais no mosto (SST, ºBrix) foram mais concentrados na CTC9003. O potencial de hidrogênio (pH), independentemente da variedade, variou de 4,00 a 4,33. A população microbiana média para leveduras apresentou valores de 10⁷ unidade formadora de colônia mililitro^-1 (UFC/mL), e para bactérias, 10³ UFC/mL. Nas características físico-químicas da cachaça de alambique, todas as variedades apresentaram concentrações dentro da faixa exigida pela legislação brasileira. As variedades CTC9002 e RB036066 podem ser utilizadas para diversificar o campo varietal das fazendas produtoras de cachaça.

Termos para indexação:
Bebida destilada; sistema de produção; fermentação; Saccharum spp.; bebida de qualidade

Introduction

Cachaça is a classic Brazilian alcoholic beverage distilled from fermented sugarcane juice. The beverage manufacturing sector, including cachaça production, generates approximately 134.6 thousand direct jobs in Brazil (Brasil, 2024). In 2023, cachaça exports from Brazil generated US$ 20.24 million, highlighting the sector’s expanding international trade potential and its growing contribution to the national economy (Brasil, 2024).

Sugarcane (Saccharum spp.), the primary raw material for cachaça production, is known for its high sucrose content and significant biomass yield (Martins et al., 2024). Sugarcane varieties intended for cachaça production must exhibit substantial stalk yield, high sucrose content, resistance to major pests and disease, ease of stripping, and extended industrial usability (Pimentel & Andrade, 2020). Therefore, selecting varieties adapted to the agro-industrial system is crucial for standardizing pot still cachaça production. However, scientific data on the impact of different sugarcane varieties on the physicochemical composition of cachaça remain limited.

Pot still cachaça farms are typically dominated by a single variety (RB867515) or sugarcane of unknown genetic origin. The repeated use of the same variety or genetically unknown materials across multiple crop seasons can reduce productivity and crop longevity (Pimentel & Andrade, 2020). These practices also increase susceptibility to biotic and abiotic stressors, further diminishing productive capacity (Cursi et al., 2022).

Existing literature on cachaça production primarily focuses on fermentation processes, particularly the juice quality (Bortoletto & Alcarde 2015; Ribeiro et al., 2017; Mutton et al., 2020) and the role of fermentative yeasts (Portugal et al., 2016; Moura et al., 2020; Stefenon et al., 2021). However, studies specifically addressing the influence of sugarcane varieties on the productivity and quality of pot still cachaça are limited (Cravo et al., 2019; Silva et al., 2020b).

Given this gap, it is essential to investigate the effects of different sugarcane varieties on the physicochemical composition of pot still cachaça under standardized cultivation, fermentation, and distillation conditions. Standardization of fermentation and distillation, combined with the use of varieties with known genetic backgrounds, is key to strengthening the cachaça production chain. This study aimed to evaluate four new sugarcane varieties (RB966928, RB036066, CTC9002, and CTC9003) and assess their impact on agronomic yield and the physicochemical properties of pot still cachaça.

Material and Methods

Field analyses

The field experiment was conducted during the 2022/23 crop year at Cachaçaria João Mendes (distillery) in Perdões, MG, Brazil (21^o03’18’’ S, 45^o00’15’’ W; altitude: 907 masl). According to Soil Taxonomy (Soil Survey Staff, 2014), the soil in the experimental area is classified as typical Hapludox. The regional climate is characterized as Cwa (monsoon-influenced humid subtropical) according to the Köppen classification, with a mean annual rainfall of 1,500 mm (Alvares et al., 2013).

Experimental design and execution

A randomized block experimental design was adopted, with five sugarcane varieties (RB867515, RB966928, RB036066, CTC9002, and CTC9003) and five replications. The variety RB867515 was used as a control due to its widespread cultivation in pot still cachaça production. Each plot consisted of four rows, each 5 m long and 1.4 m apart. Data was collected from the two center rows, excluding 0.5 m from each end. The acronyms RB and CTC represent the sugarcane genetic improvement programs in Brazil: Rede Interuniversitária para o Desenvolvimento do Setor Sucroenergético (Interuniversity Network for Sugarcane Development) and Centro de Tecnologia Canavieira (Sugarcane Technology Center), respectively.

Monthly climate data, including average precipitation (pp, mm), maximum temperatura (Tmáx, °C), and mínimum temperature (Tmín, °C), were recorded during the experimental period (planting: 29/10/2022; harvest: 29/09/2023) for the month of October/2022 (pp: 16.7 mm; Tmáx: 28.8 °C; Tmín: 18.6 °C), November/2022 (pp: 174 mm; Tmáx: 29.8 °C; Tmín: 18.6 °C), December/2022 (pp: 260 mm; Tmáx: 28.1 °C; Tmín: 17.9 °C), January/2023 (pp: 686 mm; Tmáx: 31.2 °C; Tmín: 19.5 °C), February/2023 (pp: 115 mm; Tmáx: 31.4 °C; Tmín: 19.6 °C), March/2023 (pp: 97 mm; Tmáx: 32.6 °C; Tmín: 18.2 °C), April/2023 (pp: 102 mm; Tmáx: 27.5 °C; Tmín: 17.9 °C), May/2023 (pp: 0.0 mm; Tmáx: 25.3 °C; Tmín: 11.5 °C), June/2023 (pp: 8 mm; Tmáx: 24.1 °C; Tmín: 11.5 °C), July/2023 (pp: 6.8 mm; Tmáx: 27.5 °C; Tmín: 15 °C), August/2023 (pp: 31 mm; Tmáx: 27.7 °C; Tmín: 16 °C), and September/2023 (pp: 38.9 mm; Tmáx: 34.2 °C; Tmín: 17.7 °C).

Before beginning the field experiment, soil samples were collected at depths ranging from 0 to 20 cm to determine their chemical and physical properties. The soil had a pH (H₂O) of 6.1 and contained 3.60 dag kg^-1 of organic matter (OM), 34.10 mg dm^-3 of phosphorus (P), and 208.0 mg dm^-3 of potassium (K). Exchangeable acidity (H + Al) was 2.60 cmolc dm^-3, with no detectable aluminum (Al). Calcium (Ca) and magnesium (Mg) contents were 4.70 and 0.90 cmolc dm^-3, respectively. The base saturation (BS) was 6.16 cmolc dm^-3, and the potential cation exchange capacity (CEC (T)) was 8.76 cmolc dm^-3, with zero percent aluminum saturation (m), indicating the absence of exchangeable aluminum in the soil. Micronutrient concentrations were as follows: zinc (Zn) at 1.40 mg dm^-3, iron (Fe) at 39.90 mg dm^-3, manganese (Mn) at 14.90 mg dm^-3, copper (Cu) at 0.80 mg dm^-3, boron (B) at 0.01 mg dm^-3, and sulfur (S) at 12.40 mg dm^-3. Soil texture analysis revealed a composition of 58 dag kg^-1 clay, 32 dag kg^-1 silt, and 10 dag kg^-1 sand.

The soil was prepared by disking, and nine-month-old stalk segments were planted on October 29, 2022. Fertilization at planting included 190 kg ha^-1 of monoammonium phosphate (MAP), along with 1.0 L ha^-1 of imidacloprid and 0.5 L ha^-1 of pyraclostrobin. The fertilizer and pesticides were applied at the base of the furrow before being covered with soil. Topdressing of 100 kg ha^-1 of nitrogen (urea) and 90 kg ha^-1 of K₂O (potassium chloride) was applied at 60 days after planting (DAP) (Cantarella et al., 2022). Manual weeding was performed throughout the crop cycle.

Assessment of sugarcane agronomic traits

The stalks were harvested on September 29, 2023, 330 days after planting (DAP). We collected two bundles of 10 stalks each (excluding the aerial parts) at random from the designated sample area. The maturity index (MI) and total soluble solids content (TSS, °Brix) were determined according to the methodology established by Consecana (2006).

Stalk yield (TSH, tons of stalks per hectare) was calculated using the formula (1):

$$\text{TSH = (W10S*NSM)/S}$$ (1)

Where W10S represents the weight of 10 industrializable stalks per plot, NSM denotes the number of stalks per meter, and S reflects the spacing between rows.

The expected cachaça yield (ECY) was calculated following the methodology of Pimentel et al. (2024), and liters of cachaça per ton of sugarcane (LCT) were obtained by dividing ECY by TSH.

Cachaça production

Ten stalks were randomly selected from each plot to form bundles of 50 stalks per variety. Milling, fermentation, and distillation were performed at Cachaçaria João Cassiano (distillery) in Candeias, MG, Brazil (20°45′13″ S, 45°16′53″ W; altitude: 979 masl).

The juice was extracted by grinding (10 x 14 inch mill at 4800 rpm) and filtered (60 mesh sieve) to remove coarse impurities (bagasse). Milling occurred within 16 hours after harvest. The must (diluted sugarcane juice) was adjusted to 15ºBrix, using potable, chlorine-free water, with a pH of 5.0-5.8 and temperature ranging from 28 to 32ºC. Each variety yielded 45 L of must.

Fermentation was carried out using CA-11 yeast (LNF - Leveduras Nativas de Lavras), following the manufacturer’s recommendations. The process was carried out in five 50-liter conical stainless-steel tanks, with each tank assigned to a different sugarcane must variety. Yeast separation from the must occurred through sedimentation. The must was added in five feeding stages: an initial 5.0 L, followed by four additional feedings of 10 L each at 15 °Brix, with 30-minute intervals between each addition. Fermentation was considered complete when the wine (fermented must) reached a value of ≤ 1 °Brix or when the maximum fermentation time of 24 h was reached. Upon completion, 25 L of must from each tank was diverted through a side opening for distillation using a 50-L copper still.

Only the “head” and “heart” fractions of the distillate were collected based on the initial alcoholic strength. Dias and Machado (2020) recommend collecting 1-5% of the fermented must volume as “head” if the initial distillate strength exceeds 65% vol., and 5-10% if the initial distillate strength is less than that. In this investigation, because the initial alcohol concentration was 70% vol., 1% of the total must volume was collected as the “head”. The “heart” fraction represented 12% of the total fermented must volume. Distillation was carried out in a “hot head column” 50 L still, heated by direct fire at a constant temperature of 90 ºC.

Evaluation of the must

Samples for each variety were collected one hour after the fermentation tanks were filled with 45 L of must. The evaluated parameters included total soluble solids content (TSS, °Brix), measured using a Brix saccharimeter, and pH, determined by direct measurement (Centro de Tecnologia Canavieira - CTC, 2005).

Serial dilutions (10^-1 to 10^-5 CFU/mL) were prepared and plated in triplicate to count the microbial populations at the start of fermentation. Yeast counts were determined using YEPG culture medium, composed of [(g/L): yeast extract (10), glucose (20), peptone (20), and agar (20), all from HiMedia except glucose (Dinamica), with the pH adjusted to 3.5. For bacterial counts, Nutrient Agar culture medium supplemented with 0.4% nystatin (Prati Donaduzzi) at a concentration of 28 g/L was used. The inoculated Petri dishes were incubated for 48 hours at 28 °C for yeasts and 37 °C for bacteria. After incubation, colony-forming units were counted and expressed as CFU/mL, following the methodology described by Martins et al. (2019).

Evaluation of the cachaça

The physicochemical analyses of the cachaça samples were conducted in accordance with Ordinance No. 24, dated September 8, 2005, issued by the Brazilian Ministry of Agriculture, Livestock and Supply (MAPA) (Brasil, 2005). These studies were conducted at the Quality Analysis Laboratory of Spirits in the Federal University of Lavras (UFLA). Parameters evaluated included alcohol content, volatile acidity, esters and aldehyde concentrations, higher alcohols, furfural, methanol, sec-butyl and n-butyl alcohols, ethyl carbamate, acrolein, copper content, and organoleptic attributes (Brasil, 2005).

The quantification of higher alcohols and methanol was performed using gas chromatography with a flame ionization detector (FID), following the methodologies described by Vilela et al. (2007) and Barbosa et al. (2022). We only used analytical-grade reagents and chromatography-compatible standards. Calibration curves were prepared using a 4 g/L standard solution in 40% ethanol. The chromatographic conditions included an initial column temperature of 35 °C, which was progressively increased to 140 °C. The injector and detector temperatures were set at 150 °C and 170 °C, respectively, with a carrier gas flow of 1.4 mL/min, operating in split mode with a 1:10 ratio. Ethyl carbamate was analyzed following the methodology proposed by Anjos et al. (2011).

Statistical analysis

The data on agronomic performance were first analyzed for normality using the Shapiro-Wilk test. Analysis of variance (ANOVA) was then conducted using the F-test (p < 0.05). Based on the results, Tukey’s test was applied at a 5% significance level using the Sisvar® program (Ferreira, 2019). Descriptive statistics were used to analyze the data from the fermented must (mean values) and the physicochemical characteristics of the cachaça (mean and standard deviation), based on two replicates.

Results and Discussion

Agronomic parameters of sugarcane varieties

Among the evaluated sugarcane varieties, CTC9002, RB036066, and RB966928 showed the highest total soluble solids content (TSS, °Brix). However, the TSS of RB966928 did not differ significantly from that of RB867515 and CTC9003 (Table 1).

Table 1:
Agronomic and production parameters of sugarcane varieties: total soluble solids (TSS, ºBrix), maturity index (MI), tons of stalks per hectare (TSH), expected cachaça yield (ECY), and liters of cachaça per ton of stalks (LCT).

Varieties	TSS (ºBrix)	MI	TSH (Mg ha-1)	ECY (thousand L ha-1)	LCT (L Mg-1)
RB867515	19.40 b	0.87 b	105.40 bc	10.67 ab	94.27 b
CTC9003	19.50 b	0.91 b	91.16 c	9.37 b	93.60 b
RB966928	20.20 ab	0.97 a	91.96 c	9.15 b	99.25 b
CTC9002	20.70 a	0.91 b	127.17 a	14.38 a	113.71 ab
RB036066	21.00 a	0.92 ab	113.75 ab	14.45 a	131.68 a
C.V. (%)	2.43	2.92	10.33	17.65	13.14
Overall mean	20.16	0.92	105.88	11.61	106.50

• Mean values followed by the same letter in a column do not differ significantly according to Tukey’s test (p ≤ 0.05). C.V. (%): Coefficient of variation. ECY refers to the total distillate volume and does not account for fraction separation (“head”, “heart”, and “tail”), which is relevant to marketable cachaça quality.

According to Pimentel and Andrade (2020), sugarcane varieties intended for cachaça production should have a TSS content above 18 °Brix at harvest. All varieties evaluated in this study met this requirement, indicating their suitability for cachaça production. The high °Brix values ​​observed may be attributed to favorable climatic conditions, such as milder temperatures and a period of water deficit, both of which promote sucrose accumulation (Pimentel & Andrade, 2020).

The highest maturity index (MI) values were recorded for RB966928 and RB036066, with RB036066 statistically equivalent to RB867515, CTC9003, and CTC9002 (Table 1). All varieties showed maturity indices above 0.85, which are considered adequate based on Rossetto’s (2022) scale, with values below 0.6 indicating green sugarcane and above 1.0 suggesting sugarcane in maturity decline. Sugarcane maturation is a physiological process involving the synthesis of carbohydrates in the leaves and their translocation and storage as sucrose in the stalk, specifically in the parenchyma tissues (Wang et al., 2013).

Crop maturation is determined by a complex combination of climatic variables, genetic potential, and crop management practices (Pimentel & Andrade, 2020). Stalk yield (TSH) differed significantly across varieties, with the CTC9002 variety showing the highest mean value at 127.17 Mg ha^-1, followed by RB036066 with 113.75 Mg ha^-1 (Table 1). The RB867515 and RB036066 varieties were statistically equivalent. All varieties demonstrated productivity above the average reported for the Southeast region (85.04 Mg ha^-1) and the national average (81.12 Mg ha^-1) projected for the 2023/24 crop year in Brazil (Companhia Nacional de Abastecimento - Conab, 2024). Despite these positive results, a productive and economically viable sugarcane field should meet the “three-digit” yield criterion, defined as an average yield of ≥ 100 Mg ha^-1 over five consecutive cycles. According to Landell (2017), this benchmark is typically achieved with first-cycle yields ranging from 120 to 140 Mg ha^-1, followed by a gradual decline of 5-10% in subsequent cycles. Based on this criterion, only the CTC9002 variety met the desired “three-digit” yield.

The RB036066 variety yielded the highest value for expected cachaça yield (ECY) at 14.45 thousand liters, which was statistically equivalent to CTC9002 (14.38 thousand liters) and RB867515 (10.67 thousand liters) (Table 1). However, compared to RB867515, commonly cultivated for pot still cachaça production, the ECY of RB036066 and CTC9002 was 35.4% and 34.7% higher, respectively. Moreover, the ECY of RB867515 was not significantly different from that of CTC9003 and RB966928 (Table 1). Overall, the ECY results closely mirrored the trends observed in TSS values, confirming a direct proportional relationship in which higher TSS content enables greater juice dilution during fermentation standardization (typically adjusted to 14-16 °Brix), thereby increasing ECY (Schwan & Dias, 2020; Pimentel et al., 2024). This resulted in a mean total yield of 14.45 and 14.38 thousand liters per hectare of cachaça for the varieties RB036066 and CTC9002, respectively.

Variations in TSH also affected the ECY outcomes. The varieties RB966928 and CTC9003, which had the lowest ECY values, were also among those with the lowest stalk yields. This underscores the hypothesis that high stalk yield, together with high soluble solids content, regulates both the volume of juice available for dilution and the volume of must for fermentation, factors that directly affect ECY (Medeiros et al., 2017).

Regarding liters of cachaça per ton of stalks (LCT), the highest values were observed for RB036066 (127.80 L Mg^-1) and CTC9002 (113.71 L Mg^-1), with the latter statistically similar to all other varieties. This parameter offers a general assessment of cachaça yield in liters per unit area. Although it does not specifically quantify the premium fraction known as the “heart”, it serves as an effective indicator for growers to monitor the overall productivity of their sugarcane fields. Thus, the results indicate that even with lower TSH, the total cachaça yield per hectare can be comparable to that of varieties with higher TSH, as observed in the performance of RB966928 relative to RB867515. This outcome can be attributed to factors such as higher TSS content, which allows for greater juice dilution, the variety’s vegetative growth potential, the efficiency of the agro-industrial process, and other contributing elements.

In summary, the RB036066 variety showed the highest mean values for LCT, while CTC9002 led in TSH. Both varieties also achieved the highest ECY values, reinforcing their potential for efficient and high-yield cachaça production.

Fermentation parameters in sugarcane must

Fermentation of sugarcane juice, a crucial stage in cachaça production, significantly impacts the beverage’s final quality. As shown in Figure 1, total soluble solids (TSS) content in the must varied among the evaluated sugarcane varieties. The highest TSS was observed in the variety CTC9003 (8.5 °Brix), and the lowest in RB036066 (5.0 °Brix), with an overall average of 6.30 °Brix. The pH values ranged from 4.00 to 4.33 across all varieties (Figure 1). Although this range is slightly below the optimal pH of 4.5 to 5.0 for maximum yeast activity (Agu & Oduola, 2021), it remains within the tolerance range of S. cerevisiae, which can grow at pH levels between 4.0 and 6.0 (Narendranath & Power, 2005). Maintaining slightly acidic conditions helps suppress undesirable bacterial growth, thereby minimizing deterioration during the fermentation process.

Figure 1:
pH and total soluble solids (°Brix) in the must during the first hour of fermentation, by sugarcane variety. Data were collected at Alambique João Cassiano (distillery), Candeias, Minas Gerais, Brazil. The horizontal solid line indicates the overall average pH (μpH) across all varieties, while the dashed line represents the average total soluble solids (μBrix).

Several studies highlight the efficiency of S. cerevisiae in response to pH variations. Liu et al. (2015) evaluated the effect of pH on fermentation by S. cerevisiae and described strains capable of producing ethanol across a pH range of 2.50 to 4.50. Lin et al. (2012) investigated ethanol production from sugarcane juice using S. cerevisiae BY4742 and observed that pH values below 4.0 led to the formation of acetic acid (Cardoso et al., 2022).

Maintaining an appropriate pH range is essential for successful fermentation. These conditions help prevent an extended lag phase and yeast inhibition (optimal pH 4.5-5.0). Additionally, it limits the growth of spoilage bacteria such as acetic acid bacteria, which thrive at pH ≤ 3.0, and lactic acid bacteria, active between pH 4.00 and 4.50. These bacteria can slow fermentation and deteriorate the must (Schwan et al., 2001; Liu et al., 2015; Agu & Oduola, 2021). Nevertheless, the TSS (°Brix) and pH values observed during the first hour of fermentation indicate that alcoholic fermentation, or the conversion of sugars into ethanol and carbon dioxide, began under suitable conditions. However, fermentation also depends on other factors, such as temperature, oxygen availability, raw material quality, and yeast strain (Schwan & Dias, 2020).

The mean microbial population in the must was 10⁷ CFU/mL for yeast and 10³ CFU/mL for bacteria (Figure 2). Variety CTC9002 showed the highest mean yeast count (6 × 10⁷ CFU/mL), while the highest bacterial population (5 × 10³ CFU/mL) was observed in RB867515. These yeast levels are consistent with those reported by Schwan et al. (2001), who reported populations ranging from 10⁷ to 10⁸ CFU/mL during the first hour of fermentation. Similarly, total bacterial counts in this study (≤ 10⁵ CFU/mL) align with acceptable ranges for fermenting must (Brexó & Sant’Ana, 2018).

Figure 2:
Yeast and bacterial populations in the must during the first hour of fermentation, by sugarcane variety. Data were collected at Alambique João Cassiano (distillery), Candeias, Minas Gerais, Brazil. The horizontal dashed line represents the mean yeast population (μYeast) across all varieties, while the solid line indicates the mean bacterial population (μBac).

Portugal et al. (2016) investigated microbial dynamics during sugarcane juice fermentation and observed that increasing ethanol concentrations to significant levels resulted in a decline in the populations of non-Saccharomyces species. Their findings highlight the crucial role of tolerance to abiotic stress conditions, especially ethanol, in the dominance of S. cerevisiae over non-Saccharomyces species, as further supported by Comitini et al. (2021).

In this context, careful control of both abiotic and biotic factors is essential for an efficient fermentation process. The diversity of microorganisms, along with levels of soluble solids (°Brix) and pH in the fermenting must, can significantly affect the cachaça production system in pot still distilleries.

Physicochemical parameters of cachaça quality

Physicochemical analyses of cachaça quality were carried out for each sugarcane variety (Table 2). The organoleptic assessments showed normal results. All other parameters were also within the limits established by Brazilian legislation (Brasil, 2022), except for the copper concentration in the CTC9003 variety.

Table 2:
Physicochemical analyses of cachaças by sugarcane variety, cultivated during the 2022/23 crop year. Data were collected at Alambique João Cassiano (distillery), Candeias, Minas Gerais, Brazil.

Parameters analyzed
Variety	Alcohol content (% v/v)	Organo. exam.	Secondary compounds (mg/100 mL of anhydrous ethanol)x
			Volatile aciditya	Higher alcoholsb	Aldehydesc	Estersd	Furfurale	Total congeners (a, b, c, d, e)
RB867515	45.75 ± 0.00	N	27.34 ± 2.27	315.93 ± 5.24	23.04 ± 0.00	33.96 ± 0.00	ND	400.27
RB966928	45.02 ± 0.01	N	32.68 ± 0.00	318.34 ± 4.86	22.73 ± 0.33	36.43 ± 0.00	ND	410.18
RB036066	46.88 ± 0.04	N	21.97 ± 0.02	302.75 ± 2.72	22.71 ± 0.02	35.91 ± 1.27	ND	383.34
CTC9002	46.55 ± 0.02	N	25.29 ± 0.01	308.83 ± 9.98	23.76 ± 0.29	30.60 ± 1.28	ND	388.48
CTC9003	40.15 ± 0.04	N	31.16 ± 2.63	270.02 ± 5.67	15.16 ± 0.38	19.35 ± 0.28	ND	335.69
Limits*	48.0 - 38.0	-	150.0	360.0	30.0	200.0	5.0	650.0 - 200.0
Overall mean	44.87	-	27.68	303.17	21.48	31.25	ND	383.59

• ¹mean ± standard deviation; Organo. exam. = organoleptic examination; N = normal; ^xresults corrected to mg/100 mL of anhydrous ethanol; ^aas acetic acid; ^cas acetaldehyde; ^das ethyl acetate; ^*according to Brasil (2022).

All varieties exhibited normal organoleptic characteristics (color, aroma, and flavor) (Table 2), indicating quality assurance during the distillation and fermentation processes (Vicente et al., 2006; Schwan & Dias, 2020). Effective control of the microbial community during fermentation is essential for the formation of intermediate and secondary compounds, which significantly contribute to sensory attributes (Gomes et al., 2009). In the present study, we used yeast strain CA-11 for fermentation, which is likely responsible for the typical organoleptic characteristics observed. Ribeiro et al. (2017) previously reported that CA-11 improves chemical characteristics compared to native yeasts, while Mutton et al. (2020) concluded that it produces cachaça of superior quality.

The alcohol content was highest in the RB036066 (46.88% v/v) and CTC9002 (46.55% v/v) varieties (Table 2), yet all samples remained within the standard range (38.0-48.0% v/v) established by legislation for cachaça (Brasil, 2022). Ethanol, the primary alcohol produced during alcoholic fermentation, has a high perception threshold (100 ppm) and a mild aroma, contributing little to the beverage’s overall aroma profile (Cardoso, 2020; Silva et al., 2023).

The RB036066 variety showed the lowest volatile acidity (21.97 mg/100 mL), but values across all varieties remained well within the standard limit of 150 mg/100 mL of anhydrous ethanol (Brasil, 2022). The overall mean volatile acidity in our study was 27.68 mg/100 mL of anhydrous ethanol (Table 2). Earlier, Cravo et al. (2019) reported a slightly higher mean of 31.65 mg/100 mL across different agronomic and production conditions. Acidity levels in cachaças can vary considerably due to factors such as poor hygiene, the inclusion of citrus fruit in the must during fermentation, the presence of acetic bacteria, oxygen availability, and the type of yeast used. All these factors can affect the conversion of sugar into acetic acid (Cardoso, 2020). Therefore, the results indicate that the hygiene measures adopted in this study were effective. Agronomic management also plays a role in volatile acidity. Silva et al. (2020a) evaluated pot still cachaças produced from three sugarcane varieties (RB867515, RB966928, and RB855453) cultivated under organic, conventional, and no-fertilization regimes. They reported volatile acidity values ranging from 13.58 to 23.05 mg/100 mL of anhydrous ethanol, with no significant differences among varieties or treatments. All values remained within legal limits. The five varieties examined in our study exhibited slightly higher volatile acidity, which may be due to differences in fertilization and production environments. Other factors influencing acidity include post-harvest storage duration and the type of yeast used (Cravo et al., 2019; Schwan & Dias, 2020). Therefore, standardizing practices from cultivation through fermentation and distillation may help reduce variability in acidity levels in Brazilian cachaça.

Higher alcohols are defined as the sum of n-propanol, isobutanol, and isoamyl alcohols per 100 mL of anhydrous ethanol. In our study, the RB966928 and RB036066 varieties exhibited the highest and lowest concentrations, respectively (Table 2). On average, the varieties showed values 18.74% below the maximum permissible limit of 360 mg/100 mL established by Brazilian legislation (Brasil, 2022). The formation of higher alcohols is promoted by factors such as sugarcane storage before milling, elevated fermentation temperatures, and an acidic must pH (Cardoso, 2020; Schwan & Dias, 2020). Similar levels were reported by Silva et al. (2020a) in cachaças produced from RB867515 (262.06 mg/100 mL) and RB966928 (202.77 mg/100 mL), regardless of the agronomic practices applied. Fermentation conditions, including yeast cell density, temperature, and final alcohol content, also significantly influence the concentration of higher alcohols (Ratkovich et al., 2023). In the present study, this parameter showed elevated values, though still within the maximum limit permitted for the beverage. This was primarily due to the proper separation of distillation fractions (“head”, “heart”, and “tail”), each of which contains different concentrations of alcohols (Cardoso, 2020). Therefore, producers should be aware that regulatory non-compliance most often involves excess levels of higher alcohols and volatile acidity (Lima et al., 2022).

The variety CTC9003 had the lowest total aldehyde concentration (Table 2), and all varieties remained within legislative limits (Brasil, 2022). Aldehydes such as acetaldehyde, furfural, and hydroxymethylfurfural contribute unpleasant odors and are associated with adverse sensory effects; therefore, low concentrations of these compounds are desirable in cachaça (Silva et al., 2020c). The overall mean aldehyde concentration was 21.48 mg/100 mL of anhydrous ethanol, 39.66% below the maximum limit. Silva et al. (2020a) studied various sugarcane varieties and reported an overall mean value of 24.30 mg per 100 mL of anhydrous ethanol. They attributed the higher concentrations to the type of fertilization used in sugarcane agronomic management.

Interestingly, we did not detect furfural in any of the evaluated samples (Table 1). This compound is known to impart a pungent odor when present. Santiago et al. (2015) reported a 3.5% non-conformity rate for furfural in cachaça produced in Minas Gerais. Similarly, Bortoletto and Alcarde (2015) found that 2.10% of sugarcane spirit and cachaça samples exceeded legal limits for the combined content of furfural and hydroxymethylfurfural. The absence of furfural in this study likely reflects sound production practices, such as refraining from pre-harvest burning, maintaining distillation temperatures at or below 90 °C, and minimizing solids in the fermented wine (Lima et al., 2022). Additionally, the separation of cachaça distillation fractions plays a critical role in controlling furfural concentration, as the “tail” fraction, characterized by a higher boiling point and a greater concentration of water-soluble volatile compounds, tends to accumulate more furfural (Ratkovich et al., 2023).

Our results indicate a tendency for total aldehyde accumulation in the RB867515 variety, which, along with CTC9002, exhibited the highest values. Silva et al. (2020a) also reported elevated aldehyde levels in RB867515, with 28.21 mg/100 mL of anhydrous ethanol, corresponding to 94.03% of the limit established by Brazilian legislation. Similarly, Cravo et al. (2019) found the highest concentration in this variety (26.48 mg/100 mL) under conditions comparable to those in the present study.

All varieties showed ester concentrations within the limits established by legislation (Brasil, 2022), with the lowest concentration observed in CTC9003 (Table 2). Esters are key contributors to the flavor and aroma of alcoholic beverages, with ethyl acetate and ethyl lactate being the main esters found in distilled alcoholic beverages (Gao, Fan, & Xu, 2014). The sugarcane variety has a limited impact on ester content, as its formation depends primarily on distillation and aging processes. Cachaças produced in copper stills, as in this study, tend to have higher levels of ethyl acetate and ethyl lactate compared to those distilled in stainless steel column stills (Nascimento, Cardoso, & Franco, 2008; Ratkovich et al., 2023). Aging in wooden barrels promotes the formation of aromatic esters, contributing to the development of a more complex bouquet in cachaça (Chaves & Póvoa, 1992).

In summary, analysis of secondary compounds across varieties demonstrated that RB966928 and RB867515 had the highest concentrations, primarily due to elevated levels of higher alcohols. Nevertheless, all cachaças complied with legal standards (Brasil, 2022). The varieties exhibited an overall mean concentration of 383.59 mg/100 mL of anhydrous ethanol, representing 52.13% of the minimum and 69.45% of the maximum concentration limits permitted for cachaça.

Physicochemical parameters of contaminants in cachaça

Contaminant analyses were conducted for each sugarcane variety (Table 3), with all mean values complying with the limits established by Brazilian legislation (Brasil, 2022).

Table 3:
Contaminant levels in cachaças by sugarcane variety cultivated during the 2022/23 crop year. Data were collected at Alambique João Cassiano (distillery), Candeias, Minas Gerais, Brazil.

Variety	Contaminants
	Coppera	Methyl alcoholb	Sec-butylic alcoholb	N-butylic alcoholb	Ethyl carbamatec	Acroleinb
RB867515	2.35 ± 0.32¹	<0.19* ± 0.00	<0.48 ± 0.00	<1.22 ± 0.00	8.04 ± 0.04	<0.48 ± 0.00
RB966928	3.63 ± 0.25	<0.19 ± 0.00	<0.48 ± 0.00	<1.22 ± 0.00	8.81 ± 0.17	<0.48 ± 0.00
RB036066	2.70 ± 0.08	<0.19 ± 0.00	<0.48 ± 0.00	<1.22 ± 0.00	9.05 ± 1.44	0.50 ± 0.00
CTC9002	2.93 ± 0.02	<0.19 ± 0.00	<0.48 ± 0.00	<1.22 ± 0.00	8.77 ± 0.64	<0.48 ± 0.00
CTC9003	7.22 ± 0.00	<0.19 ± 0.00	<0.48 ± 0.00	<1.22 ± 0.00	9.99 ± 0.65	<0.48 ± 0.00
Upper limit**	5.0	20.0	10.0	3.0	210.0	5.0
Overall mean	3.77	0.19	0.48	1.22	8.93	0.48

• ¹mean ± standard deviation; ^aconcentration in mg/L; ^bresults corrected to mg/100 mL of anhydrous ethanol; ^cconcentration in μg/L; *DL = detection limit; **Brasil (2022).

The highest copper concentration was detected in the CTC9003 variety, likely due to inefficient cleaning of the copper still, as distillation began with this batch (Table 3). Proper sanitation practices, such as thoroughly cleaning the still and coils and filling them with water to minimize copper oxidation, play a crucial role in lowering copper contamination in cachaça (Cardoso, 2020). Therefore, copper content in cachaça is related to still sanitation rather than the sugarcane variety itself.

Methanol (methyl alcohol) was below detection limits in all samples (Table 3), highlighting the effectiveness of proper sugarcane juice treatment. It forms during fermentation through the enzymatic breakdown of pectin-rich sugarcane fragments (bagasse) (Cardoso, 2020; Ratkovich et al., 2023). Methanol ingestion above legal limits can cause severe acidosis, respiratory complications, coma, or even death (Maia & Campelo, 2006). Silva et al. (2020a) reported methanol levels of 3.42 and 3.30 mg/100 mL anhydrous ethanol in the RB867515 and RB966928 varieties, respectively, indicating limited varietal influence on this contaminant.

Levels of n-butyl and sec-butyl alcohols were also below detection limits (Brasil, 2022). These alcohols are typically produced due to contamination by the bacterium Clostridium acetobutylicum during fermentation (Santiago et al., 2020). Silva et al. (2020a) reported n-butanol concentrations ranging from 0.96 to 1.20 mg/100 mL anhydrous ethanol in the RB867515, RB966928, and RB865453 varieties, regardless of management practices. Low levels of n-butyl and sec-butyl alcohols can be maintained by minimizing bacterial contamination. This includes avoiding sugarcane storage near animal stables or milking areas and refraining from using stored sugarcane (Maia & Campelo, 2006; Cardoso, 2020).

Ethyl carbamate (EC) was within the legal limits for all varieties (Brasil, 2022), with an overall mean value of 8.92 μg/L, corresponding to just 4.25% of the maximum permissible value. This potentially carcinogenic compound, commonly found in distilled beverages, is primarily formed from urea, which may be naturally present in sugarcane juice or improperly added to the must. Its formation can also involve cyanide ions released through the enzymatic degradation of cyanogenic glycosides in sugarcane (Silva et al., 2020b; Lima et al., 2022). D’Avila et al. (2016) reported variability in EC levels among cachaças from RB867515 under different fermentation systems, production environments, and agronomic management practices, with values ranging from 12.29 to 31.37 μg/L across Minas Gerais municipalities. Cravo et al. (2019) also observed a wide concentration range (4.47-49.48 μg/L) of EC in cachaça under varying agronomic and production practices. These findings underscore the need to standardize agro-industrial processes, from cultivation through distillation, to minimize EC variability.

Acrolein was below detection thresholds for all varieties except RB036066 (Brasil, 2022). This aldehyde can form during cachaça distillation through glycerol dehydration or via the action of lactobacilli in the must, which convert glycerol into β-hydroxypropionaldehyde (Ratkovich et al., 2023). Masson et al. (2012) evaluated seventy-one samples of sugarcane spirits from small- to medium-sized stills and reported that 9.85% of the samples had acrolein levels exceeding the limits established by Brazilian legislation. In a study by Ribeiro et al. (2017) comparing the effects of natural yeast and selected yeast (CA-11) on the composition and quality of distillates from the SP83-2847 variety, acrolein was detected only in beverages fermented with natural yeast, though still within legal limits. Similarly, in the present study using the same selected yeast (CA-11), acrolein was detected in small amounts only in the RB036066 variety, supporting the conclusion that process standardization contributes to higher-quality cachaça.

Overall, all varieties met the legal physicochemical contaminant standards, confirming that proper production and sanitation practices were effectively implemented.

Conclusions

Among the evaluated sugarcane varieties, RB036066 and CTC9002 demonstrated superior agronomic performance. Notably, RB867515 continues to play an essential role in the cachaça agroindustry due to its stable performance and adaptability. All cachaças produced from the tested varieties complied with the physicochemical standards set by Brazilian legislation. These findings provide scientifically validated alternatives to the predominant use of RB867515 or unidentified genotypes, contributing to a more diversified and resilient base for cachaça production in Brazil.

Acknowledgments

The authors thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) - Funding Code 001 and the Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG) for support in carrying out the present study.

Data Availability Statement

Data available upon request to authors.

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• Editor de seção:
  Renato Paiva

Publication Dates

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

History

• Received
  07 May 2025
• Accepted
  18 Aug 2025