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Evaluation of physical-chemical and microbiological

characteristics and sensory profile of

pilsner beers sold in Serra Gaúcha

Nayara Monique Lazzari Grolli1, Natália Inês Cavagnolli2, Cristian Roncada3, Diego Broch4, Deise Pretto5,

Indiara Ceriotti Bombana6, Patrícia Kelly Wilmsen Dalla Santa7, Daniel Alexandre González Massens8, Queli

Defaveri Varela9.

DOI: https://doi.org/10.35699/2447-6218.2024.45167

Abstract

Objective: To evaluate and compare physicochemical, microbiological, and sensory characteristics of Pilsen beers

marketed in Serra Gaúcha. Methods: In the first study, 11 samples, with the Pilsen denomination on the label, were

submitted to physicochemical, microbiological, and organoleptic tests. In the second comparative, five people were

identified and selected as appreciators (Group A) and five people were identified and selected as non-appreciators

(Group B). Results: The item flavor showed statistical significance between Group A and the reference beer. In the

physicochemical analyses, there was a statistical difference (p≤0.01) in color between the reference and Group B,

which also presented a higher than average alcohol content as compared to the others. As for the microbiological

analyses, one sample presented growth of Brettanomyces sp. It is therefore possible to verify that deficient standards

exist in the manufacturing process of the products where the inclusion of additives is allowed, regardless of origin,

which ultimately confuses and hinders the consumers’ perceptions of quality in the product. Conclusion: More ri-

gid legal parameters are suggested and new production techniques are needed to generate more differentiated and

higher-quality products. Finally, a clear legal deficiency was observed in the brewing sector.

Keywords: Beer, Pilsen, brewing industry.

Avaliação das características físico-químicas e microbiológicas e perfil sensorial de cervejas

pilsen vendidas na Serra Gaúchs

1Centro Universitário da Serra Gaúcha - FSG. Caxias do Sul, RS. Brasil.

Orcid: 0000-0001-9706-2184

2Universidade de Caxias do Sul - UCS. Caxias do Sul, RS. Brasil.

Orcid: 0000-0001-6920-320X

3Pontifica Universidade Católica do Rio Grande do Sul - PUCRS. Porto Alegre, RS. Brasil.

Orcid: 0000-0003-3782-4911

4Centro Universitário da Serra Gaúcha - FSG. Caxias do Sul, RS. Brasil.

Orcid: 0000-0002-3365-2529

5Centro Universitário Metodista - IPA. Porto Alegre, RS. Brasil.

Orcid: 0000-0001-9723-6375

6Universidade de Caxias do Sul - UCS. Caxias do Sul, RS. Brasil.

Orcid: 0000-0002-8488-8961

7Universidade de Caxias do Sul - UCS. Caxias do Sul, RS. Brasil.

Orcid: 0000-0002-9101-8534

8Centro Universitário da Serra Gaúcha - FSG. Caxias do Sul, RS. Brasil.

9Universidade de Caxias do Sul - UCS. Caxias do Sul, RS. Brasil.

Orcid: 0000-0002-8488-8961

*Autor para correspondência: nayaraintx@gmail.com

Recebido para publicação em 09 de março de 2023. Aceito para publicação 19 de setembro de 2023.

e-ISSN: 2447-6218

Grolli, N. M. L. et al.

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

Resumo

Objetivo: Avaliar e comparar as características físico-químicas, microbiológicas e sensoriais das cervejas Pilsen comer-

cializadas na Serra Gaúcha. Métodos: No primeiro estudo, 11 amostras, com a denominação Pilsen no rótulo, foram

submetidas a testes físico-químicos, microbiológicos e organolépticos. Na segunda comparação, cinco pessoas foram

identificadas e selecionadas como apreciadoras (Grupo A) e cinco pessoas foram identificadas e selecionadas como

não apreciadoras (Grupo B). Resultados: O item sabor mostrou significância estatística entre o Grupo A e a cerveja

de referência. Nas análises físico-químicas, houve diferença estatística (p≤0,01) na cor entre a referência e o Grupo

B, que também apresentou um teor alcoólico acima da média em comparação com os outros. Quanto às análises

microbiológicas, uma amostra apresentou crescimento de Brettanomyces sp. É possível verificar que existem padrões

deficientes no processo de fabricação dos produtos onde a inclusão de aditivos é permitida, independentemente

da origem, o que acaba confundindo e dificultando as percepções dos consumidores sobre a qualidade do produto.

Conclusão: Parâmetros legais mais rígidos são sugeridos e novas técnicas de produção são necessárias para gerar

produtos mais diferenciados e de maior qualidade. Por fim, observou-se uma clara deficiência legal no setor cervejeiro.

Palavras-chave: Cerveja, Pilsen, indústria cervejeira.

Introduction

The beer market in Brazil has shown significant

growth, and the structures of the beer industry are highly

concentrated and characterized by the presence of leading

companies and brands. According to data obtained by

the Brazilian Craft Beer Association, Abracerva, in 10

years, Brazil has gone from 70 to 700 craft breweries

with regularized registration, as shown in the graph in

Figure 1 (Ramos, 2019).

Figure 1 – Number of Craft Breweries in Brazil

Source: MAPA (2018)

The macro-breweries, which serve the entire

national territory, are formed by the Medium and Lar-

ge Brewing Companies (MGE) and are responsible for

the largest share of beer consumption among the popu-

lation and the largest production in terms of volume.

The microbreweries, which are incorporated into the

Micro and Small Brewing Companies (MPE), respond

to preliminary local demand, and are characterized by

low production volumes and lower installed production

capacity. However, as published by MAPA in 2020, 204

new breweries were registered in Brazil and another 30

canceled their registrations, which represents an increase

of 174 breweries, 14.4% compared to the previous year

(Sgorla, A. F., 2023).

The current definition of beer production in Brazil

follows rules defined by legislation, which specifies that

beer originates from the alcoholic fermentation of wort,

derived from barley malt and drinking water and the

action of yeast intermingled with hops (SEBRAE, 2017).

Evaluation of physical-chemical and microbiological characteristics and sensory profile of pilsner beers sold in serra gaúcha

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

Any modification to this process and in the raw material

can result in beers with different physicochemical and

sensory characteristics. In mid-2014, a survey conducted

by the Barth Haas Group showed that Brazil is the third

largest beer producer worldwide behind only China and

the United States, having produced approximately 14.04

billion liters in 2014. Years later, Brazil is still the third

largest beer producer in the world (Ramos, 2019; Lima

J. D. M., 2023).

The amount of alcohol depends very much on the

fermentation process. The production and consumption of

the majority beer in Brazil comes from lager pilsener, which

is a low fermentation beer fermented by S. pastorianus.

This yeast is a hybrid of S. cerevisiae and S. eubayanus.

It is characterized by mild flavor, pH around 4.3, clear

and limpid (Pereira, 2020). During fermentation, the

sugars transform into alcohol, heat and CO2 , a combi-

nation that results in a carbonated alcoholic beverage

(De Arruda, Junior and Goulart, 2013). In Brazil, Pilsen

beer is less full-bodied, with reduced bitterness and low

alcohol content, thus being lighter and more refreshing.

Such measures are adopted as standard by major natio-

nal brands, thus performing a mixture of styles between

Pilsen, Lager and Lager Beer (Beltramelli, 2012; Scheffer

et al. , 2013).

Historians believe that craft beer has existed

for centuries when bread was fermented in water (Bel-

tramelli, 2012). Presently craft beers are judged to be

part of a class of products derived from superior quality

and higher added value. There are elaborate means of

formulation and processes distinct from those products

created on an industrial scale. Small-scale production

enables the development of differentiated products for the

most exquisite consumers, with emphasis on valuing the

selection of regional raw materials for product creation.

(CERVBrasil, 2019).

For the characterization and identification of the

considerable compounds for beer quality, physicoche-

mical and sensory methods are widely applied. Sensory

methods are grouped into three streams: affective, dis-

criminative, and descriptive tests. Affective tests aim to

objectify whether the product is accepted or preferred

by the consumer. In discriminative tests, researchers

analyze if the subjects can discern a perceptible difference

between the products. In descriptive tests, researchers

seek to determine the main relevant points between the

products studied and their intensities (Dutra, 2019).

Given the growth in consumption in the sector

and the large number of brewing companies, the study

is necessary to evaluate the physicochemical, microbio-

logical, and sensory characteristics, aiming to assess the

products available to consumers in Serra Gaucha.

Methods

This is a comparative analytical study, performed

in two stages. The first study was elaborated with 11

Pilsen beers: five of the craft type, five of the industrial

type, and one from the Czech Republic. The products were

purchased between November 1, 2016 and November

5, 2016 in local commerce. The main inclusion criterion

the characterization of Pilsen type beer on the respective

labels.

The sensory evaluation was performed through

a descriptive method, which establishes qualitative and

quantitative differences in each type of beer. Evaluations

are performed using attribute and proportion scales.

Odor and taste, texture, appearance, flavor and texture

properties are described. The relationship between the

duration and intensity of a stimulus is measured.

It was performed by ten individuals of both

sexes, aged between 22 and 38. Of the group members,

five had a university degree in oenology, with technical

knowledge of brewing. The remaining five, considered

self-proclaimed connoisseurs, had no formal training in

the area.

The sensory analysis was performed through tas-

ting in an appropriate room with all the necessary items

for the tests. The samples were uncharacterized, wrapped

in a black bag in order to make it impossible to identify

their commercial brand, and served one by one to each

participant, with approximately 100mL at a temperature

of 2°C in a glass suitable for tasting. After tasting each

beer, the participants filled out a beer evaluation form,

adapted from BJCP Beer Scoresheet, 2009, which was

collected at the end of the tastings.

In the physical-chemical analysis, the procedures

for pH, volatile acidity, turbidity, color, density, alcohol,

total dry extract and total acidity were performed. We used

methodologies from the Beverage and Vinegar Operatio-

nal Manual (IN 24/2005), from the alcoholic fermented

products section, being the methods two, three, four, six

and eight for density, alcohol, pH, volatile acidity and total

dry extract, respectively. For total acidity, turbidity and

color analysis, methods MA-AS3113-01 from the World

Organization of Vine and Wine, OUGH C.S, Methods

Analysis of Musts and Wines and Analytica EBC Method

9.6 were consulted, respectively.

The samples were also submitted to three mi-

crobiological tests: identification of aerobic mesophilic

microorganisms, by the Plate Count Agar technique, Com-

pedium of Methods for the Microbiological Examination

of Foods; mold and yeast, by the DRBC Agar technique

- APHA 4th Edition, Compedium of Methods for the Mi-

crobiological Examination of Foods; and Brettanomyces sp.

by the plating/selective medium technique - Millipore .

Grolli, N. M. L. et al.

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

In the second study, 10 beers were purchased, 5

craft Pilsen and 5 industrial Pilsen. Ten individuals were

invited to participate in the tasting, five of whom were

beer lovers, i.e., individuals who maintain the habit of

consuming beer with a certain frequency, and five others

who were not beer lovers. In an appropriate room, 10

samples of totally uncharacterized beers were placed

so that the individuals could identify, through tasting,

whether the beer was craft or industrial, through sensory

analysis of the product. The same descriptive method was

applied.

In order to ensure a reliable result, the analyses

were performed with the help of a laboratory accredited

by the CGCRE, with vast experience in beverage analysis.

To conclude the results, it was necessary to apply the

T-test for mean and standard deviation. The software

used for statistical analysis was SPSS 20.0 for Windows.

Results and Discussion

The results of the physicochemical analyses of

the beers are presented in tables 1 and

2. The evaluation of the sensory profile is pre-

sented in the form of a table, numbers 3 and 4. We can

observe that there were no significant results among the

factors analyzed (p<0.05). The analyses, provided by a

laboratory with INMETRO/Cgecre accreditation, were

made after calibrating the equipment, as well as reading

the control sample, with previously known values, and

the “blank” sample, in order to verify the accuracy and

precision of the results.

Table 1 – Physical-chemical analyses of beers from macro breweries (Group A), beers from micro breweries (Group B)

and beer from the Czech Republic (B6)

Samples pH Volatile Acidity

(meq/L) Turbidity

A1 4,62 0,276 00,60

A2 4,35 0,246 00,41

A3 4,32 0,246 00,86

A4 4,19 0,198 01,06

A5 4,37 0,294 00,69

B1 4,65 0,240 59,70

B2 4,47 0,366 03,60

B3 4,86 0,192 19,90

B4 4,79 0,186 13,20

B5 4,63 0,114 29,20

B6 4,75 0,190 01,80

Mean ± SD 4,55 ± 0,218 0,37 ± 0,125 11,91 ± 18,562

Values expressed as Mean ± Standard Deviation (SPSS 20.0 for Windows); * p<0.05.

In the samples analyzed, five beers, four industrial

and one handmade, showed pH values below 4.5, which

makes them unsuitable for consumption as pH values up

to this range are of fundamental importance, because they

inhibit the development of pathogenic microorganisms,

especially Clostridium botulinum, the bacterium that

causes botulism, and thus prevent further contamination.

Beer contamination can occur due to several factors, and

in addition to the performance of laboratory tests, in

many breweries sensory tests are also performed (Nunes,

2021).

The acidity in the product is due to the formation

of acids during fermentation; the main organic acids

formed are: acetic acid, formic acid, pyruvate, D-lactate,

L-lactate, among others. Thus, it is of utmost importan-

ce to perform a strict quality control to guarantee the

standardization of the beer and avoid losses and waste

during the production process (Nunes, 2021). Regarding

microbiological analysis, only one microbrewery sample

showed growth of Brettanomyces sp. with 1 CFU/mL.

In the second test, composed of a group of lovers

and a group of non lovers, we can see that there was also

no significant difference between them, as shown in table

Evaluation of physical-chemical and microbiological characteristics and sensory profile of pilsner beers sold in serra gaúcha

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

Table 2 – Physical-chemical analyses of beers from macro breweries (Group A), beers from micro breweries (Group B)

and beer from the Czech Republic (B6)

Samples Color (EBC

Units) Density at 20°C Alcohol

(% vol.)

Dry Extract

(g/L)

Total Acidity

(meq/L)

A1 7 1,00830 4,56 38,3 22,10

A2 5 1,01000 4,71 43,4 19,50

A3 6 1,00950 4,70 42,1 21,20

A4 5 1,00950 4,40 41,1 17,00

A5 7 1,00768 3,85 34,5 17,40

B1 7 1,01243 4,80 49,6 18,90

B2 10 1,01287 5,13 51,9 24,40

B3 12 1,00840 4,74 39,4 16,20

B4 7 1,01040 4,83 44,7 16,40

B5 12 1,01076 4,62 44,7 21,10

B6 10 1,00890 4,82 40,1 16,40

Mean ± SD 7,75 ± 2,541 1,01 ± 0,002 4,65 ± 0,322 42,71 ± 4,980 19,14 ± 0,276

Values expressed as Mean ± Standard Deviation (SPSS 20.0 for Windows); * p<0.05.

Table 3 – Comparison of sensory analyses of winemakers between beers produced in macro breweries (Group A), beers

produced in micro breweries (Group B) and beer from the Czech Republic (B6)

Criterion

Group A Group B Group B6

Average Standar d

Deviatio

Average Standar d

Deviatio

Average Standar d

Deviatio

Appearance 2,92* 0,76 3,84* 0,75 3,20 1,79 <0,01*

Aroma 4,40* 2,25 6,68* 1,55 5,60 3,21 <0,01*

Flavor 4,92* 2,22 6,24 2,33 7,20* 1,48 0,04*

Palate 3,08 1,50 3,80 1,41 3,40 2,07 0,25

Values expressed as Mean ± Standard Deviation (SPSS 20.0 for Windows); * p<0.05.

Table 4 – Comparison of sensory analyses of non-enologists between beers produced in macro breweries (Group A),

beers produced from micro breweries (Group B) and beer from the Czech Republic (B6)

Criterion

Group A Group B Group B6

Average Standar d

Deviatio n Average Standar d

Deviatio n P

Appearanc e 3,56 0,87 3,80 0,96 4,00 0,71 0,49

Aroma 7,26 1,79 7,04 2,92 6,20 3,56 0,69

Flavor 7,10 1,19 7,16 2,54 8,20 1,30 0,50

Palate 3,48 1,19 3,20 1,55 3,20 1,92 0,77

Values expressed as Mean ± Standard Deviation (SPSS 20.0 for Windows); * p<0.05.

Grolli, N. M. L. et al.

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

Table 5 – Differentiation of industrial craft beers into connoisseurs (Group A) and non-connoisseurs (Group B)

Group A (Appreciators) Group B

(Non-Appreciators)

Hits Errors Hits Errors

Total 35 15 31 19

Values expressed as Mean ± Standard Deviation (SPSS 20.0 for Windows); * p<0.05.

Thus, the groups had difficulties in differentia-

ting craft beer from industrial beer. In view of this, it is

possible to see a lack of standard in the manufacturing

of products, where the use of various additives is allowed

or not, stages such as mashing, fermentation, maturation

and the use of different ingredients such as wheat, corn,

rye, rice, honey, fruit and others that can change the taste

of the product. Regardless of the origin, which ends up

masking the organoleptic characteristics, confusing and

damaging the consumers’ taste.

The old German Beer Purity Law (Reinheitsge-

tbot), dating back to 1516, states that beer should be

produced without any adjuncts, exclusively with malt,

hops and water. Brewer’s yeast should not be considered

as a raw material, being necessary only as a biochemical

agent in the transformation of sugars into alcohol by the

fermentation process. This law established the quality of

the ingredients, not the manner in which they should be

processed. (Dias, 2018).

The physicochemical criteria are important ins-

truments used by the legislation as a way to control and

evaluate the production processes as seen in Tables 1

and 2. Since it influences many factors, such as microbial

growth, color intensity, enzyme activity, oxy-reduction

potential, and also flavor, it should be kept below 4.5

(Dutra, 2019).

The alcohol content was higher in the microbre-

wery groups, with an average of 4.82%, and in the beer

of Czech origin, with 4.65%. Regarding color, according

to Article 38 of Decree No. 6871 of June 4, 2009 (Dutra,

2019), beer can be classified as light beer, which has a

color corresponding to less than twenty EBC units (Eu-

ropean Brewery Convention). All analyzed samples were

classified as light, characteristic of Pilsen beers. A study

that analyzed the information on the labels of Pilsen beers

found that all the brands studied were in accordance with

current labeling standards (Almeida, 2015).

Since beer is defined as the favorite among Brazi

lians (IBOPE, 2013), it would be expected that beer lovers

would identify different characteristics when compared

to products from micro or macro breweries, as well as

when compared to a beer from the Czech Republic, the

birthplace of Pilsen beer. The lack of identification of

differentiated sensory characteristics among beer lovers

demonstrates the need for improvements in the perceptive

capacity of this group, which can often end up consuming

a product through advertising trends (Della Lucia et al. ,

2013), price, or even the maturity of sensory evaluation.

Through the evaluation of the sensory profile by

the groups, one can see that higher education in the area

is important in the differentiation of parameters such as

appearance and aroma, mainly. A similar study observed

that, when comparing two Pilsen type beers produced

in micro breweries with two market-leading Pilsen type

beers, the micro breweries’ beers have greater intensity in

the sensory attributes, differentiating themselves in color,

yeast aroma, sweet taste and bitter taste. However, there

was no statistical difference between these evaluators,

contrary to what was expected, because micro breweries

develop a beer with more artisanal characteristics and

that should provide greater attraction to the consumer

(Minim e Da Silva, 2016).

For further testing, regarding technical matching

for differentiating samples appropriately about consumer

perception, the Check-All-That-Apply (CATA) method

can be used. It involves the use of agreeability scales as

well as the characteristics attributes to differentiate a

product. The supplied scale to consumers can be used to

design an internal preference map, in which it is possible

to visualize consumer response towards products. Using

narrative attributes provided by a trained panel, along

with the consumer’s preference map, can be used to create

an external preference map, which can help understand

which attributes are important in relation to consumer

preferences (Lee, 2023).

Conclusion

Regarding the microbiological evaluation, of the

eleven samples used, ten were not contaminated, while

the missing sample showed the presence of Brettanomyces

sp. with 1 CFU/mL; probably the presence of this yeast

in the sample was not able to influence the sensory cha-

racteristics of the samples, however, its presence should

be monitored because this microorganism is the main

responsible for causing turbidity, odors, and undesirable

flavors in the final product.

Even though the microbrewery market is on

the rise and their products are reaching a larger portion

of the beer consumption in Brazil, they must explore

characteristics of the production process that aim to

positively surprise the consumers’ sensory evaluation.

As proof of this, during the two tests performed with the

Evaluation of physical-chemical and microbiological characteristics and sensory profile of pilsner beers sold in serra gaúcha

Cad. Ciênc. Agrá., v. 15, p. 01–07, DOI: https://doi.org/10.35699/2447-6218.2024.45167

group without technical knowledge (which represents the

largest part of the beer consuming population) they were

unable to identify the differences between a craft product,

characterized by the use of higher quality raw materials,

and an industrial product, commonly known for using

other raw materials in order to lower production costs.

Thus, these data provide us with the understanding that

the choice of product occurs by factors that go beyond the

organoleptic experience and are closer to an economic,

cultural, visual issue, by indication of friends or family,

and mainly by the influence of the media. During the

development of this research, the legal deficiency in the

brewing industry was also identified, since there is little

legislation regarding manufacturing and quality control,

causing difficulty in production standardization and ge-

nerating gaps in the effectiveness of product surveillance.

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