Jurnal Akademika Kimia, 14(2): 83-89, May 2025
ISSN (online) 2477-5185 | ISSN (print) 2302-6030
http://jurnal.fkip.untad.ac.id/index.php/jak/

OPEN ACCESS

1

Protein Content and Organoleptic Test Central Sulawesi Pumpkin Seed
Tempeh (Cucurbita moschata) for Learning Resource
Raodatul Jannah, *Tri Santoso, Sri H. V. Pulukadang, & Dewi S. Ahmar

Chemistry Education Study Program/Faculty of Teacher Training and Education – Universitas Tadulako, Palu –
Indonesia 94119
Received 19 March 2025, Revised 31 March 2025, Accepted 08 April 2025
doi: 10.22487/j24775185.2025.v14.i2.pp83-89

Abstract
The yellow pumpkin (Cucurbita moschata) is a plant that readily adapts to diverse soil conditions; however, the utilization of its
seeds remains restricted. Despite its relatively high protein content, which is crucial for growth and energy, it can serve as an alternative
raw material for tempeh production. This study aims to generate yellow pumpkin seed tempeh in compliance with SNI 3144:2015,
assess organoleptic variations in the protein content of yellow pumpkin seed tempeh from three distinct regions in Central Sulawesi, and
produce a Student Worksheet as an alternative resource for chemistry education, validated by expert lecturers. This research employs a
quantitative methodology utilizing the Kjeldahl analysis technique to ascertain protein concentrations across three phases: destruction,
distillation, and titration. Furthermore, a sensory evaluation was conducted by 20 panelists to analyze the texture, color, flavor, and
scent of yellow pumpkin seed tempeh. The resultant tempeh has favorable organoleptic properties, achieving an average preference score of
4. A uniform white coloration, a distinctive tempeh aroma, and a dense texture characterize it. Protein content research revealed that
tempeh from Palu City had the highest protein concentration at 24.5%, followed by Palolo District at 23.91% and North Lore District
at 20.41%. This suggests the impact of environmental factors on the nutritional composition of the manufactured tempeh. According to
SNI 3144:2015, the visual characteristics of yellow pumpkin seed tempeh and its protein content conform to the established quality
criteria. The findings of this study were presented as Student worksheets, which achieved a validity rate of 85%, indicating their high
suitability as a learning resource. This research enhances the application of yellow pumpkin seeds as a nutritious food source and fosters
the creation of engaging and novel educational materials.

Keywords: Kjeldahl, organoleptic, protein, pumpkin seed tempeh, student worksheets

Furthermore, protein can be an alternative
energy source when carbohydrates and fats are
Pumpkin is an annual crop that is easy to insufficient for the body's needs. This process
grow and widely distributed in Indonesia. It has high involves breaking down proteins into components
production in Java, 270.000 tons per year. Sumatra
that can be used to produce energy, thus helping the
94.000 tons per year, and Bali 70.000 tons per year
body function properly in meeting energy
(BPS, 2021). Despite its potential, the community's deficiencies. Thus, pumpkin seeds, rich in protein,
use of it is still limited to fruit due to a lack of can be an ideal ingredient for making tempeh, which
knowledge about its nutritional content (Pabidang is not only delicious but also has high nutritional
et al., 2020). Along with the development of various
value and is part of Indonesian culinary culture
innovations, pumpkin seeds have gained attention (Indonesian National Standard). The protein
as a material with great potential for full utilization. content of pumpkin seeds makes them a potential
This potential comes from the bioactive alternative material for making tempeh, a typical
compounds' antioxidant properties inside pumpkin Indonesian fermented food. Through the
seeds. This compound stops the formation of free
fermentation process of soybeans using the
radicals from peroxide and prevents the oxidation Rhizopus oligosporus fungus, tempeh is created,
of unsaturated fatty acids in cell membranes. So it is which is nutritious and favored by the wider
suitable for health, especially in reducing community. Based on the Indonesian National
inflammation and protecting the body's cells from
Standard (2015), some of the criteria for tempeh
serious diseases. These benefits include the that need to be considered are having a compact
presence of amino acids, zinc, magnesium, various
texture, meaning that when cut, the tempeh remains
essential fatty acids, vitamin C, carotenoids, sterols,
intact and does not fall off easily, the color is evenly
and cryptoxanthin (Mumpuni & Khasanah, 2021).
white, and it has a distinctive aroma of tempeh
⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯

Introduction

*Correspondence:
Tri Santoso
e-mail: trisantoso@untad.ac.id
© 2025 the Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons
Attribution-NonCommercial-ShareAlike 4.0 International, which permits unrestricted non-commercial use, distribution,
and reproduction in any medium, provided the original work is properly cited.

83

Raodatul Jannah et al.
without the smell of ammonia. Therefore,
researchers tried to utilize pumpkin seeds as a
substrate in making tempeh.
Previous research has shown that tempeh
from pumpkin seeds has a high protein content of
24.66 % and is suitable for use as a learning medium
(Hasrina & Nurdin, 2021). Adding pumpkin seed
powder has also been shown to affect the chemical
properties of tempeh (Mahfud et al., 2021). This
research aims to produce pumpkin seed tempeh in
accordance with the organoleptic standards of
tempeh based on the Indonesian National Standard
(2015), this is carried out based on consideration of
the appearance of important tempeh and the main
thing in public acceptance, especially in ensuring
that yellow pumpkin seed tempeh products have
good quality and acceptable, especially this study
researches tempeh with substrate innovation in the
environmental conditions of plant growth that are
influenced by nutrients that are absorbed by plants
to continue growing, where these nutrients come
from the soil related to plant nutrition so it needs to
be researched. In addition, the results of this
research are expected to be developed into student
worksheets as an alternative source for learning
chemistry. Student worksheets are a medium that
provides learning stimuli to students so that they
can think critically and be motivated in learning
(Hanik, 2020).
This research is important because the quality
of tempeh, both in terms of nutrition and
appearance, significantly affects public acceptance.
At the same time, the environmental conditions in
which plants grow also determine their nutritional
content. Based on the description above, this study
aims to produce yellow pumpkin seed tempeh in
accordance with the Indonesian National Standard
(2015), organoleptically test the differences in
protein content of yellow pumpkin seed tempeh
from three different regions in Central Sulawesi, and
create student worksheets as an alternative source
for chemistry learning.

protein content test materials, including tempeh
yellow pumpkin seeds, NaOH 30 %, Kjedhal
tablets, concentrated H2SO4, HCl 0.1 N, NaOH 0.1
N, methyl red, and aquadest.

Sample preparation

Yellow pumpkin seed samples were taken
from three different regions in Central Sulawesi:
Palu City (Tinggede Village), Sigi (Bahagia Village),
and North Lore (Watutau Village). Hasrina &
Nurdin (2021), the sample preparation process
began with separating the pumpkin seeds from the
fruit, then washing them thoroughly, boil for 10
minutes, drain and separate the yellow pumpkin
seeds from the skin, rewash the yellow pumpkin
seeds with clean water, and drain again until there is
no water, mix 0.1 grams of Raprima yeast of the
microbial type Rhizopus sp. with 100 grams of yellow
pumpkin seeds until evenly distributed, put the
mixture of Raprima yeast and yellow pumpkin seeds
in plastic, cover the tempeh packaging plastic with
fire to cover the plastic, punch a hole in the tempeh
packaging with a needle and let it sit for 48 hours.

Analysis of tempeh protein content

Nuryanti et al. (2024) analyzed the protein
content in pumpkin seed tempeh samples using a
modified Kjeldahl method. The destruction begins
by weighing 0.5 grams of the sample, placing it into
a 100 mL Kjeldahl flask, and then adding 1 Kjeldahl
tablet and 10 mL of concentrated H2SO4 solution.
The sample is then destroyed until the solution
becomes clear. After the solution has cooled, the
distillation stage is carried out by diluting the
solution resulting from the destruction process with
100 mL of aquades. Next, 5 mL of the sample is
taken and put into the distilled tube, then 10 mL of
30 % NaOH solution is added through the wall of
the distilled tube. The distillation process is
regulated by a condenser and an Erlenmeyer flask
containing 10 mL of 0.1 N HCl solution and 3 drops
of methyl red indicator. At the titration stage, the
distillate solution is titrated with NaOH 0.1 N until
the color changes to a clear yellow, indicating the
titration endpoint. Make a blank solution using a
similar procedure. However, the sample was
replaced with aquades.

Method

Tools and materials
The tools used in this study are divided into
tempeh-making tools, including basins, pots, Rinnai
RI-201S gas stoves, scales, sieves, spoons, and
toothpicks. Tempeh protein testing equipment
includes Sonic Electronic analytical scales, distillates,
Pyrex burettes 50 mL, sticks and clamps, 5 mL and
10 mL Iwaki measuring cups, 100 mL Pyrex Iwaki
measuring cups, 50 mL and 100 mL Pyrex chemical
cups, 5 mL Pyrex volume pipettes, 10 mL Pyrex glass
funnels, 100 mL Kjedhal flasks Pyrex, 100 mL Iwaki
pyrex Erlenmeyer, drip pipettes. Moreover, the
ingredients in this study are divided into ingredients
for making tempeh, including yellow pumpkin
seeds, water, tempeh yeast, and plastic, as well as

Organoleptic testing

Organoleptic testing of yellow pumpkin seed
tempeh was conducted in three regions in Central
Sulawesi to assess differences in product preference
based on the panelists' reactions to the samples
provided (Purbasari & Putri, 2021). The
organoleptic test was carried out by 20 untrained
panelists who were students from the Chemistry
Education Study Program. Samples of yellow
pumpkin seed tempeh were given to the panelists.
Each sample was provided raw, and the panelists
were asked to fill out a questionnaire with a scale of
product preferences on five indicators: color,
aroma, texture, and taste, as shown in Table 1.

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Jurnal Akademika Kimia

Table 1. Scale and description of organoleptic test assessment
Rating
Scale
1
(very
much
dislike)
2
(dislike)
3
(just
likes)

Description of Assessment
Color

Smell

Texture

Taste

Gray/brownish
mycelium

Rotting/stinging/smelli
ng of ammonia

Soft/easily crumbles/not
fused

Very acidic/bitter

Not uniform
/partly yellowish

Not fresh/ sour smell

Less compact/easy to
separate

Slightly acidic/less
typical tempeh

Fairly white/less
even

Smell not strong/
slightly bland

A bit compact/a little
brittle

A little bland/still
acceptable

Compact/not easily
destroyed

Typical tempeh/not
sour/ bitter

Very compact/dense/
stays intact when cut

Very tasty/typical
tempeh/savory/not
sour/ bitter.

4
(likes)

Flat white/ no
black stains

5
(very
like)

Clean white/
uniform/no
stains

Typical aroma of
tempeh/ no pungent
smell
Fresh/distinctive/stron
g/ aroma of tempeh
without sour or
ammonia smell

Creation and validation of student worksheets

Information:
Vs
= Sample Titration Volume
Vb
= Volume of Blank
14
= Atomic Weight of Nitrogen
N.HCl = Normality HCl
6.25
= Conversion Factor
P
= Dilution

The initial phase involves creating the
Learner Worksheet as a resource for chemistry
education. Material and design specialists validate
the Student Worksheet to ascertain its suitability as
a chemistry learning resource.
The evaluation of the eligibility and validity
of the Student Worksheet employs quantitative
descriptive calculations utilizing the following
method.
𝑟𝑒𝑠𝑢𝑙𝑡 𝑠𝑐𝑜𝑟𝑒𝑠
Percentage = 𝑡𝑜𝑡𝑎𝑙 𝑣𝑎𝑙𝑖𝑑𝑎𝑡𝑖𝑜𝑛
× 100%
ℎ𝑖𝑔ℎ𝑒𝑠𝑡 𝑠𝑐𝑜𝑟𝑒

Analysis of organoleptic test data
The evaluation of organoleptic test results involved
determining the feasibility % of yellow pumpkin
seed tempeh products across various assessment
criteria, including color, aroma, texture, and flavor,
utilizing the specified formula.

(1)

(Sugiyono, 2016)

Average =

The eligibility percentage for Student Worksheets is
analyzed according to the categories outlined in
Table 2.

Source: Pattimura et al. (2020)

Protein content

The findings of the protein content tests of
the yellow pumpkin are shown in Table 3.

Interpretasi
Very valid
Valid
Quite valid
Invalid
Very invalid

Table 3. Protein yield data tempeh yellow pumpkin
seeds
No.
Sample
Protein Content (%)
Average
Tempeh
(%)
Yellow
I
II
III
Pumpkin
Seeds
1.
Palu City
24.5 26.25 22.75
24.5

Analysis of protein content
The assessment of protein levels was conducted by
Hasrina & Nurdin (2021), utilizing the subsequent
formula.

2.
3.

𝑉𝑠−𝑉𝑏 × 𝑁.𝐻𝐶𝑙 × 14 × 𝑃
%𝑁 =
× 100%
𝑚𝑔 𝑠𝑎𝑚𝑝𝑒𝑙

(2)

%Protein = % N × 6.25

(3)

(4)

Results and Discussion

Table 2. Percentage of student worksheet
eligibility
Percentage Product
Validity (%)
80 ≥ Score ≤ 100
60 ≥ Score ≤ 80
40 ≥ Score ≤ 60
20 ≥ Score ≤ 40
0 ≥ Score ≤ 20

𝑡𝑜𝑡𝑎𝑙 𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑎𝑚𝑜𝑢𝑛𝑡
𝑡𝑜𝑡𝑎𝑙 𝑝𝑎𝑛𝑒𝑙𝑖𝑠

Lore
Utara

10.5

24.5

26.25

20.41

Palolo

21

24.5

26.25

23.91

The protein content of yellow pumpkin seed
tempeh was determined using the Kjeldahl analysis
method. The Kjeldahl method is a chemical analysis
85

Raodatul Jannah et al.
technique used to ascertain the nitrogen content of
a sample. This method relies on converting nitrogen
in proteins into ammonia through a destructive
process involving sulfuric acid and a catalyst. The
resulting ammonia is then quantitatively analyzed
using sample titration. The Kjeldahl method was
chosen to analyze the protein content in tempeh
made from yellow pumpkin seeds (Cucurbita
moschata) because it is a precise, standardized, and
widely used method in food analysis. Hasrina &
Nurdin (2021) showed that the protein content in
pumpkin seed tempeh samples was lower than
24.66%. Mahfud et al. (2021) recorded a protein
content of 23.07 % in tempeh with the addition of
yellow pumpkin seed powder. The ability of this
method to measure total nitrogen in fermented
foods makes it very reliable, even if the samples are
complex or non-homogeneous. In addition, this
method can be applied to a small number of
samples, making it efficient for laboratory research
purposes and the preparation of learning media
based on laboratory test results.
The protein content of yellow pumpkin seed
tempeh, as indicated in Table 3, varies across
different regions in Central Sulawesi. The sample
from Palu City exhibited the most significant
protein level at 24.5 %. Hasrina & Nurdin (2021)
indicated that the protein level in the yellow
pumpkin seed tempeh sample was inferior to 24.66
%. Lathifah & Hermawati (2019) showed that the
protein content of soybean tempeh is 31.85%,
which exceeds that of pumpkin seed tempeh. Based
on SNI 2015, the three tempeh samples have
satisfied the established protein quality standards,
which require a minimum of 15 % protein content.
The variation in yield is influenced by other
factors, including the volume of titrant required to
reach the point of equality, which is indicated by the
change in color from purple to yellowish (Yurida et
al., 2013). The varied growth location of yellow
pumpkin fruits is another factor; Although this
plant can thrive in diverse conditions, different
environmental conditions in three regions in
Central Sulawesi affect soil nutrition, thus affecting
the nutritional value of yellow pumpkin fruit. The
nutritional composition of yellow squash is
influenced by its variety, harvest maturity, and
environmental conditions (Tamer et al., 2010).
Optimal environmental conditions promote
plant growth, including soil quality, water
availability, altitude, and light intensity. Three
research locations in the Central Sulawesi region
were observed at varying altitudes: Palu City at an
altitude of 0 - 700 meters above sea level (BPS,
2025), North Lore Regency at about 1000 meters
above sea level (Widyatmoko et al., 2024), and
Palolo Regency at an altitude of 500 - 900 meters
above sea level (BPS, 2020). This shows the North
Lore and Palolo District are highland areas,
resulting in lower air temperatures and light
intensity. As a lowland, Palu City experiences high
air temperatures and light intensity. In Indonesia,
temperature variations are determined by altitude.

Altitudes below 700 meters above sea level usually
experience high temperatures, and altitudes above
700 meters generally experience cooler
temperatures. The height of a site above sea level
affects the air temperature and light exposure
received by plants (Istiawan & Kastono, 2019).
Tedianto's (2012) research underlines that
environmental conditions affect the nutrient
absorption of pumpkin plants, suggesting that an
increase in the growth area of yellow pumpkin
correlates with a decrease in its biochemical content.
Altitude affects temperature, leading to reduced
sunlight and increased CO2 concentration, which
inhibits photosynthesis and affects the protein
composition of plants. Research shows that high
soil pH correlates with an increase in the protein
content of yellow squash. Nevertheless, increased
soil moisture correlates with a reduction in protein
content. Hermita et al. (2017) revealed similar
findings, indicating that taro leaves at 800 meters
above sea level had lower protein levels compared
to those at 400 meters above sea level. This is due
to the height. The intensity of the received light
decreases, and the humidity significantly increases at
higher altitudes.
The soil's pH value is a crucial determinant
of its appropriateness for plant growth, as it
influences nutrient availability. An inadequate soil
pH can precipitate numerous issues, including
nutrient deficits, diminished microbial activity,
reduced yields, and impaired soil health. The soil pH
in Palu City ranges from 7.45 to 8.05, indicating that
it is neutral to slightly alkaline (Bakri et al., 2016).
The pH level of the soil typically varies from 3.5 to
10. Regions with considerable precipitation exhibit
a natural soil pH between 5 and 7, whereas arid
regions display a pH range of 6.5 to 9 (Taisa et al.,
2021).

Organoleptik test

The results of the research obtained
regarding the organoleptic testing of yellow
pumpkin seed tempeh include the criteria of color,
aroma, texture and taste of tempeh for the panelists,
the protein content of yellow pumpkin seed tempeh
for three regions in Central Sulawesi, including in
Palu City, North Lore District, and Palolo District
were determined using the Kjedhal method in 0.5
grams of tempeh samples and its use as a source of
chemistry learning in the form of Student
Worksheets. The following are the results of the
organoleptic tempeh submission in Table 4.
The findings of organoleptic tests indicate
that yellow pumpkin seed tempeh possesses
favorable sensory attributes and can potentially be a
delectable and high-quality food product. The
panelists' evaluation indicates a strong preference
for the yellow pumpkin seed tempeh, as its
completely white surface signifies the effective
growth of the mycelium fungus Rhizopus
oligosporus, resulting in high-quality goods.
Wihandini et al. (2012) indicated in their
study that the characteristic aroma of tempeh is
generated by mold enzymes, which decompose soy

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protein and fat into a unique scent during
fermentation. This aroma received favorable
evaluations from panelists, thereby demonstrating
the efficacy of the fermentation process.
Table 4. Results of the organoleptic test of yellow
pumpkin seed tempeh
Sample
Palu
City
North
Lore
District
Palolo
District

Test
criteria
Color
Smell
Texture
Taste
Color
Smell
Texture
Taste
Color
Smell
Texture
Taste

Average
panelist
rating
4.2
4.4
4.3
4.3
4.5
4.4
4.5
4.5
4.35
4.4
4.3
4.35

Overall
average

Rating
scale

4.3

Likes

4.5

Likes

4.3

Student worksheets are assessed by expert lecturers
who serve as validators for both content and design
elements. The validation results for the material
element indicated that the prepared student
worksheets were valid, achieving an average validity
score of 80%. In contrast, the design aspect received
an average validity score of 90%, categorized as very
valid. The validation outcomes by materials and
design specialists are displayed in Table 5.
Table 5. Validation assessment by experts
Assessment aspects
Rating scale (%)
Material Expert
80
Design Expert
90
Sum
170
Average
85

The validator's evaluation indicates that the
student worksheets require improvement based on
the
recommendations
provided.
The
recommendations and contributions offered serve
as a basis for modifying student worksheets. The
recommendation for validating the material aspect
is to modify the sampling location to align with the
background of the student worksheets.
Recommendations
and
enhancements
are
illustrated in Figure 1.

Likes

The flat and dense surface of tempeh is
caused by molds that produce hyphae and
mycelium, thus forming a compact structure
(Cempaka et al., 2020). The density of tempeh
increases with fermentation time due to loss of
water content and tissue formation in soybeans,
resulting in favorable evaluations from panelists
(Kinanti, 2024). The assessors ultimately
appreciated the yellow pumpkin seed tempeh flavor,
which resulted from the enzymatic breakdown of
the substrate's macromolecules, leading to a unique
and palatable taste (Zhao et al., 2018). The
outcomes of organoleptic evaluations indicate that
yellow pumpkin seed tempeh possesses the
potential to be a palatable and high-quality food
product, serving as an alternative for individuals
seeking healthy and nutritious dietary options. The
judges favored yellow pumpkin seed tempeh
products in the order of samples from the North
Lore District, Palolo District, and Palu City.
Nonetheless, the judges' evaluation indicated a
preference for yellow pumpkin seed tempeh. The
Indonesian National Standard (2015) evaluation for
yellow pumpkin seed tempeh indicates that it meets
the established organoleptic criteria. The tempeh
exhibits a uniform white surface, a characteristic
aroma devoid of ammonia, a compact texture that
resists disintegration, and the distinctive flavor
associated with tempeh.

a
b
Figure 1. Suggestions and improvements to the validity
of material aspects
Description: a). suggestion by material aspect validator;
b). improvement of student worksheets

The average evaluation by professionals
indicates that these student worksheets are highly
suitable as a chemistry learning resource, achieving
a rating of 85%. Student worksheets are considered
legitimate if the percentage score is greater than or
equal to 61 %. The appropriateness of the material
in the feasibility component is evaluated based on
the alignment between the presented challenge and
the learning objectives. The validation results
indicate that the arrangement of students in
addressing difficulties in worksheets is highly
effective. Issues can be resolved by extracting
information from many sources and performing
analysis. The chemistry study is intrinsically linked
to solving chemistry problems using scientific

Student worksheets

The Student Worksheet is an educational
resource that facilitates independent information
comprehension and encourages students to actively
solve problems. It is anticipated that these
worksheets will enhance students' skills. This can be
accomplished by supplying high-quality student
workbooks. One method is to validate student
worksheets (Desmiwati et al., 2017).
The validation of student worksheets
assesses their overall practicality (Sari, 2023).

87

Raodatul Jannah et al.
methodology. Consequently, educational tools
emphasizing problem-solving are essential for
cultivating students' capacity to generate new
knowledge and facilitate learning (Wahyuni &
Miterianifa, 2019).

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quality criteria of low calorie pumpkin
dessert. Notulae Botanicae Horti Agrobotanici
Cluj-Napoca, 38(1), 76-80.

Conclusions

This research shows that yellow pumpkin
seed tempeh complies with national standards for
its appearance in the three regions of Central
Sulawesi, with a panelists' preference rating of 4 in
the likes category. Protein content samples from
Palu City exhibited the highest protein content at
24.5%, followed by Palolo District at 23.91%, and
the lowest protein levels were found in North Lore
District at 20.41%. The research findings, validated
by materials and design specialists, are viable as a
chemistry learning resource, achieving an 85 %
approval rate. This research shows the potential of
yellow pumpkin seeds as a source of protein. It
produces valid chemistry learning resources to
increase awareness of the importance of using local
resources.

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