Phytochemical, Haematinic and Antidiabetic Test of Pumpkin

Extract (Cucurbita moschata)

Nova Lusiana

, Linda Prasetyaning W

, Eva Agustina

, Risa Purnamasari

, Mei Lina Fitri

Kumalasari

, Estri Kusumawati

, Esti Novi Andyarini

, Irul Hidayati

, and Agustin Nur Wildati

Universitas Islam Negeri Sunan Ampel Surabaya, Indonesia

Keywords: Phytochemical, Haematinic, Antidiabetic, Cucurbita moschata

Abstract: Pumpkins contain natural phenolic compounds which are potential antioxidants and have a bioactivity as

medicines. These compounds can be found in stems, leaves, flowers, and fruits. Pumpkin (Cucurbita

moschata) is one of the antidiabetic and antihyperglycemic traditional food which can be utillized as

substances for enteral formula. The polysaccharide and pectin contents are claimed to reduce blood glucose

levels and control glycemic levels. The purpose of this study is to determine the effect of skin, mesocarp and

pumpkin seeds (Cucurbita moschata) on hemoglobin and glucose levels. The study is an experimental study

conducted at a laboratory with 24 male BALB/c mice which were given a methanol extract of mesocarp, seeds

and pumpkin skin for 14 days in the integration laboratory of Sunan Ampel State Islamic University. Data

analysis used the Anova One Way test. The results of the study indicate that there are differences in

hemoglobin levels in each test group (p-value = 0,015). The LSD follow-up test indicates that the mesocarp

and seed groups and the control and seed groups are two significantly different groups. However, there is no

difference in glucose levels in each test group (p-value = 0,171).

1 INTRODUCTION

Pumpkins contain alkaloid compounds useful for

stimulating the nervous system, increasing or

decreasing blood pressure, and controlling microbial

infections. Natural phenolic compounds of pumpkins

are potential antioxidants and bioactivity as medicine.

This compound can be found in stems, leaves,

flowers, and fruits. In human body, flavonoid

functions as an antioxidant, protection of cell

structures, increasing the anti-inflammatory activity

of vitamin C and antibiotics (Erfanur, 2014).

Pumpkins as accessible local food contain protein

for consumption, high level digestibility (99%), beta-

carotene (1,569 μg / 100g.), and Carotenoid

particularly β-carotene, β-cryptoxanthin, lutein, and

zeaxanthin (Rodriguez 2008). Carotene is known as a

potential absorber of singlet oxygen species (ROS).

The consumption of antioxidants for instance

carotenoids, polyphenols, and tocopherols can

prevent oxidative stress. (Tourniarire, 2009). The

feeding of pumpkin flour to diabetic rats for 4 weeks

significantly reduces insulin and blood glucose levels

compared to the control group (Sedigheh, 2011).

Cucurbita moschata seed or kuaci (in

Indonesian), is not only a tasty snack, but it also has

a prevention effect of benign prostate enlargement.

The seed also contains mineral elements: Zn (zinc)

and Mg (magnesium), which are very important for

the health of the reproductive organs, including the

prostate. 100 g of Cucurbita moschata seeds contain

6.5 mg of Zn minerals (Widowati et al., 2008). The

other contents in 100 g of pumpkin seeds are calories

515.00 cals, protein 30.60 g, fat 42.10 g,

carbohydrates 13.80 g, sugar 5.30 g, calcium 54.00

mg, phosphorus 312.00 mg, iron 6.20 mg, and water

5,90 g

Pumpkins contain alkaloid compounds which are

useful for stimulating the nervous system, increasing

or decreasing blood pressure, and controlling

microbial infections. It contains natural phenolic

compounds which are potential antioxidants and have

a bioactivity as medicines. The compound can be

found in stems, leaves, flowers, and fruits. Flavonoids

in the human body function as antioxidants,

protecting cell structures and increasing the activity

of anti-inflammatory vitamin C and antibiotics

(Erfanur, 2014).

Lusiana, N., Prasetyaning, L., Agustina, E., Purnamasari, R., Kumalasari, M., Kusumawati, E., Andriyani, E., Hidayati, I. and Wildati, A.

Phytochemical, Haematinic and Antidiabetic Test of Pumpkin Extract (Cucurbita moschata).

DOI: 10.5220/0008906800002481

In Proceedings of the Built Environment, Science and Technology International Conference (BEST ICON 2018), pages 103-107

ISBN: 978-989-758-414-5

103

The purpose of this study is to determine the

phytochemical content of pumpkin and the effect of

skin, mesocarp and pumpkin seeds (Cucurbita

moschata) on hemoglobin and glucose levels.

2 METHOD

This research is experimental laboratory research

employing quantitative method that fulfils the entire

requirements to test the cause and effect relationship.

This study controls the relevant conditions of the

situation studied. The following step is observation of

the effect of manipulated conditions. In this study,

manipulated variables are extracts of some parts of

the pumpkin fruits (Cucurbita moschata), for

instance the skin of fruits, mesocarp, and seeds of the

fruit. The next step is evaluation of the most

significant part of pumpkin fruits on hemoglobin and

glucose level as a dependent variable.

The sample of this study is the extracted pumpkin

fruit (Curcubita moschata) which was extracted by

the metanol maceration process. The experimental

animals used were male mice (Mus musculus). The

extracting process of fruit pulp, skin, and seeds parts

having constant weight was macerated by methanol-

water (4:1) at temperature (25-28°C) for 10 hours.

The maceration results were then filtered with 41

whatman paper and the filtrate was centrifuged for 10

minutes. In addition, the solvent evaporated using a

rotary evaporator at a temperature of 65°C results

condensed liquid of skin of fruits, mesocarp, and

seeds of the fruit extracts

Extracted liquid per oral is provide by needle

sonde at a 2 g / kg BW and a volume of 0,2 ml for 14

days. The adding of pumpkin extract to male mice

(Mus musculus) is carried out as follows: the mice are

grouped into four groups. Group I is a control group

which is received aquades, Group II is the group

which received the pumpkin bark extract, Group III is

a group , which provided mesocarp extract pumpkin,

Group IV is a group which provided an extract of

pumpkin seeds. The research was conducted at the

integrated laboratory of UIN SunanAmpel Surabaya.

3 RESULT AND DISCUSSION

In this study,extracts of skin, mesocarp, and seeds

were tested by phytochemical tests. The tests are as

follows:

3.1 Salkowski Test

The Salkowski test was conducted to determine the

presence of sterols and triterpenoids. The initial step

of this test was scaling a 0.5 gram sample. After

scaling, the sample was added to a test tube and then

added 1 ml of chloroform; it was then homogenized

and filtered them. After the filtrate was obtained, the

3-5 drops of H2SO4 was added on its filtrate. The end

of process was observation of sterol on filtrate. If

filtrate’s color is red, it indicates the existence of

sterol. Contrarily, golden yellow on filtrate indicates

the presence of triterpenoids.

Due to the colour conversion to golden yellow, the

results of observation indicate that the mesocarp

positively contained triterpenoid. Meanwhile, skin of

fruits and seeds of the fruit did not contain

triterpenoid and sterol.

3.1 Saponin Test

Saponin test was conducted by adding 5 ml aquades

to 0.5 g the sample. If there is a foam, the sample

contains positively saponin. The observation process

indicate that the foam was formed. This foam

indicates that the mesocarp and skin test contains

saponin while the seeds do not contain saponin.

3.2 Wagner Test

In the Wagner test, 10 ml of HCl 2M was added to the

sample before the heat process for 5 minutes.

Subsequent cooling treatment on room temperature

and filter process were then conducted successively.

Filtrate as a result of filter process was then added

with 5 ml of HCl 2M and 3-5 of wagner liquid.

Sedimentation on the filter process indicates the

contents of alkonoid compounds on the extract. The

wagner test shows that pumpkin skins, seeds,

mesocarp does not contain alkanoid.

3.3 Flavonoid Test

Flavanoid test began with the weighting of 0.5 g of

pumpkin extract and filling it into a test tube.

Sufficient methanol liquid and 3-5 FeCl3 then were

added into the similar tube before conducting

observation of the colour change. If the color of the

mixture of pumpkin extracts with the liquids turns

into green, red, purple, blue and black, these

conditions indicate the flavanoid on the extract. The

observation result of the skin, the mesocarp and

pumpkin seeds find the color change to black

indicates positive flavanoid on the extract.

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3.4 Carbohydrate Test

The samples were diluted with methanol, 2 ml of the

sample, then were mixed with 2 drops of molisch

reagent until the color became brick red. Then,

H2SO4 was added and the forming of purple ring was

observed. Due to the absence of purple ring

formation, carbohydrate compounds were absent of

the skin, the mesocarp and pumpkin seeds.

3.5 Protein Test

The samples were diluted with methanol; 2 ml of the

sample was then mixed with 1 drop of NaOH and 1

drop of CuSO 4 successively. Positive results indicate

the formation of a purple ring or reddish

discoloration. From the results of the observation,

there is no color change in brick red on skin,

mesocarp or seeds.

Figure 1: figure of pythochemicals test on the skin

Figure 2: figure of pythochemicals test on seeds

Figure 3: figure of pythochemicals test on the mesocarp

Based on qualitative phytochemical tests, the

pumpkin skin methanol extract contained several

chemical compounds such as saponins and

flavonoids. In addition, the methanol extract of

pumpkin mesocarp contains chemical compounds

such as triterpenoid, saponin, and flavonoids.

Moreover, pumpkin seed extract has flavanoid

compound. This result is consistent with the research

of El Adawi TA and Taha KM (2001), which

indicates that the pulp of the pumpkin fruit or

mesocarp has bioactive components such as

polysaccharides, proteins, peptides, para-

aminobenzoic acid. A component of phenol and

sterols does not contain saponins, steroid and

triterpenoid tannins. However the component

contains secondary metabolites of flavonoids. The

results of this study show that pulp of the pumpkin

fruit or mesocarp contains triterpenoids and saponins.

Based on Pabesak et al’s study (2013), flavonoid

compounds in natural phenolic compounds have

bioactivity as a medicine (drug) and can potentially

be used as an excellent antioxidant for cancer

prevention. The total phenolic content in the pumpkin

was 93 μg GAE / g.

Based on Raharjo et al’s study (2011), pumpkin

seeds contain nutrients such as amino acids, Zn

(zinc), Mg (magnesium), essential fatty acids, vitamin

E (tocopherol), carotenoids, steroids, cryptoxanthin,

monocyclic sesquiterpenoid and trypsin inhibitors.

Trypsin inhibitors could effectively inhibit peroxide,

turning into free radical and oxidize unsaturated fatty

acids in cell membranes. This evidence supports our

results that methanol extract of pumpkin seeds

contains flavonoid compounds which have a potential

as an antioxidant. Several essential nutrients

contained in various types of pumpkin extract which

have been studied are antioxidant, antidiabetic, anti-

obesity, antyhiperkolestrolemik, anti bacterial, and

anti-prostate cancer.

The content of pumpkin polysaccharides

macromolecules consists of galactose, glucose,

arabinose, xylose and glucuronic acid which has

chemical and pharmacological contents to support

significantly pumpkin nutrition. Pumpkin fruit

polysaccharides have shown high levels of insulin in

the blood serum, reduced glucose levels in the blood,

glucose tolerance improvise. Therefore, it can be

developed as an oral antidiabetic agent (Chang et al

2014). Pumpkin polysaccharides can also inhibit the

action of preoksida hydrogen, and as the result, it

decreases cell viability. The appearance of lactate

dehydrogenase and the formation of

malondialdehyde indicate that the polysaccharide

from the pumpkin has some benefits as a

cytoprotective and antioxidant. This is consistent with

the results of our study which showed no significant

difference between skin, mesocarp and pumpkin

seeds (Cucurbita moschata) group in increasing

blood glucose levels in male mice.

The average results of measurement of the

hemoglobin level in the mice fed by mesocarp

extracts, skin and pumpkin seeds can be seen in

Figure 4.

Phytochemical, Haematinic and Antidiabetic Test of Pumpkin Extract (Cucurbita moschata)

105

Figure 4 : Average of hemoglobin level

From Figure 4, the lowest mean hemoglobin (Hb)

level in the pumpkin seed treatment group is 12.7 and

the highest mean Hb level is the treatment group

pumpkin mesocarp extract at 19

The average or mean of glucose levels in the

control group such as mesocarp extracts, skin and

pumpkin seeds are illustrated in Figure 5.

Figure 5: Average of glucose level

Based on the figure 5, the average of the lowest

glucose levels is the treatment group which are given

pumpkin seed extracts in the amount of 107,7 while

the higher glucose level in the group receiving the

pumpkin skin extract. Pumpkin has been utilized as

anti-diabetic substance and components of bioactive

compounds concentrated in the fruit (Behera, et al.,

2012). Polysaccharide extracts of cucurbita moschata

which were tested in the enzyme reaction lead to

significant differences in non-competitive inhibition

of alpha-glucosidase at a concentration of 0.7-0.9

mg/ml. It has also been shown that the powder of the

species Cucurbita moshata duch has hypoglycemic

compounds for patients with type 2 diabetes. Chen et

al, (1994) reported that because of the polysaccharide

content in humans. (Xiong and Cao, 2001, Norfezah,

et al., 2011). According to Li et al’s research, 2005,

the mechanism of anti-diabetic pumpkin is derived

from the activities of protein-bound polysaccharides

which has been proven to reduce blood glucose

concentrations, increase blood levels of serum insulin

and improve tolerance to glucose in the alloxan-

induced rat which destroys cell β and therefore,

induce diabetes. It is believed that this effect may be

due to the antioxidant properties of polysaccharides

that protect cells ß of the pancreas.

Data on hemoglobin and glucose levels are

normally distributed. The significance of hemoglobin

is 0.950, which is greater than α (0.05). Similarly, the

significance of glucose is 0.637, which is greater than

α (0.05). Therefore, testing with Anova One can be

continued because data needs must be normally

distributed. Data on hemoglobin and glucose levels

are homogeneous. The significance of hemoglobin is

0.300, which is greater than α (0.05). Similarly, the

significance of glucose is 0.126, which is greater than

α (0.05). Therefore, testing with Anova One can be

continued because requirement of homogenous data

are fulfilled

Based on the Anova One Way test, there were

significant differences in glucose levels in the Mus

Muculus group, with the mesocarp, seed and pumpkin

skin extract in the control group. This is reinforced by

the p-value (Sig.) The glucose level is 0.171, which is

greater than α (0.05). However, there were significant

differences in hemoglobin levels in the MusMuculus

group in which mesocarp, seeds, and pumpkin extract

were given to the control group. This is reinforced by

the value of p (Sig). The hemoglobin level is 0.015,

which is less than α (0.05).

The calculation of the multiple comparisons leads

to the conclusion that there are at least 2 pairs of

different groups. At the glucose levels, skin and seed

groups show a significant difference (p-value =

0.042). Meanwhile, at the hemoglobin levels, the

groups of mesocarp and seeds show a significant

difference (value of p = 0,003). These results are

similar to the control group and the seeds which also

have a significant difference (value of p = 0.012).

The different parts of the plant have also been

used as medicine in developed and developing

countries. The leaves are used as antianemic,

analgesic, and in some cases, have been used

externally for the treatment of burns and other related

diseases (Fu et al., 2006). It supports our findings that

there is a slight improvement in the hemoglobin

extract of the skin treatment group, pumpkin flesh

(mesocarp), and pumpkin seeds than those of the

contol group. The increase of hemoglobin levels

shows that pumpkin extract has haemanitic activity

on experimental animals. Research testing noodles

enriched with inorganic , organic iron only, or

combination with steamed yellow pumpkin as a

fodder for mice has been conducted to evaluate the

significant effects of yellow pumpkin to increase

hemoglobin levels. The results show that enriched

noodles with a combination of inorganic iron and

steamed pumpkin have a significant role as a food

vector and intervenes the iron deficiency that causes

anemia in mice. In the research of Widowati et al.,

(2003), pumpkin fruit is found to contain 0.071 mg of

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vitamin B6, riboflavin 0.128 mg, 18.792 mg of folic

acid and 0.147 mg of copper (Cu), per 100 g.

Pumpkin also contains carotenoids activity of 1569 g

equivalent to 261.5 Vitamin A in the form of a dry

powder containing glycine amino acid from 19.97 to

27.12 mg/100 g. Thus, in this study, it is possible that

the micronutrient-micronutrient mechanisms help

utilization of iron in the group provided by the

pumpkin extract in hemoglobin synthesis. Thus, the

level of hemoglobin in the group increases.

4 CONCLUSIONS

Pumpkins contain trypenoids, saponins, and

flavonoids. There were differences in hemoglobin

levels in each test group (p-value = 0,015). The LSD

follow-up test indicates that the mesocarp and seed

groups, as well as the control and seed groups, are two

different groups. However, there was no difference in

glucose levels in each test group (p value = 0.171).

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