Determine the cucrbita maxima (PUMPKIN SEEDS) having antioxidant and anti bacterial activity are not.

1. Presented by
Muni raja j
IV B- Pharmacy II Sem,2014 Admitted Batch
Under the guidance of
MR. PRANABESH SIKDAR
Department Of Pharmaceutical Chemistry
SEVEN HILLS COLLEGE OF PHARMACY
Accredited ‘A’ grade by NAAC
Tirupati 517561.
ANTIOXIDANT AND ANTIBACTERIALACTIVITY OF
HYDROALCOHOLIC EXTRACT OF SEEDS OF
Cucurbita maxima
1

2. SL.NO Title
1 Introduction
2 Aim and Objectives
3 Literature review
4 Plan of work
5 Materials and Methods
6 Result and Discussion
7 Conclusion
8 References
9 Appendix
Contents
2

3.  Cucurbita maxima (PUMPKIN) belongs to family Cucurbitaceae.
 It is widely cultivated by indigenous peoples throughout
Mexico, Central, and North America before the arrival of
Europeans.
 The color of pumpkins derives from orange carotenoid
pigments, including beta-cryptoxanthin, alpha and beta
carotene.
INTRODUCTION
3

4. Many farmers see pumpkins as a profitable opportunity.
Because of this, the acreage under pumpkin cultivation has
expanded greatly in recent years and competition in the
pumpkin market is increasing
 Pumpkins can be grown on most soil types, providing these
are well drained and are of good fertility
 grow best in soil with a pH between 5.5 and 6.5
 The average yield of pumpkin is between 20 to 30 tonnes
/ha.
4

5. To determine the Antioxidant and Antibacterial activity of the
hydroalcoholic extract of the seeds of Cucurbita maxima.
OBJECTIVES
Collection of the Cucurbita maxima seeds.
Standardization of the Cucurbita maxima powdered seeds.
Extraction of Cucurbita maxima seeds with hydroalcoholic
solvent.
Phytochemical screening.
In-vitro activities.
AIM
5

6. 1. Sengupta Rupa et. al. (2013) has under gone comparative studies on’’ anthelmintic potential of cucurbita maxima(pumpkin) seeds
and caricapapaya(papaya)seeds’’. The crude extracts of them were assayed against adult earth worms(Pheretima posthuma) for the
evaluation of antihelmintic activity.The method used to perform antihelmintic activity was ghosh method. Various concentrations of
these both cucurbita maxima(pumpkin) seeds and carica papaya(papaya)seeds extracts were tested and results were expressed in
terms of paralysis and death of. He concluded that both cucurbita maxima(pumpkin) seeds and carica papaya(papaya)seeds have
antihelmintic activity where carica papaya(papaya)seeds have more antihelmintic activity then cucurbita maxima(pumpkin).
2. A.B. Abd EI-Aziz et. al.(2011) has undergone studies on ‘’Antimicrobial proteins and oil seeds from pumpkin (Cucurbita
moschata)’’. They evaluated The nutritive value and biological activity of pumpkin (Cucurbita moschata) seeds cultivated in Egypt.
They conducted chemical analysis on fiber, protein, ash, carbohydrates, and fatty acids present in the non irradiated and irradiated
seeds. They found that seeds are rich in oil (44.45±2.83 %) and the oil consists of appreciable amount of unsaturated fatty acids
(71.10±4.32 %) and found to be a rich source of linoleic acid (52.64±0.90 %). Gamma irradiation of pumpkin increased significantly
(P<0.05) the yield of free fatty acid, acid value and peroxide value of extracts. Results showed decreases in the iodine value after
irradiation at doses up to 10kGy. some of the oils used on the basis of their reputed antimicrobial properties have well documented in
vitro activity . The seed of pumpkin has pharmacological activities such as anti-diabetic, antifungal, anti infllamation activities and
antioxidant effects.
3. Marimuthu amutha et. al. (1998) has undergone studies on “Determination of secondary metabolites, ld50 value and antioxidant
activity of seed extract of cucurbita pepo linn’’. They utilized HPLC and GC to analyze various photochemicals of methanolic seed
extract of C.pepo. Male Sprague Dawley rats were utilized for their studies . Male Sprague Dawley rats were administered with
MSCP at a dose of 200, 500, 1000, 2000, 2500 and 3000 mg/kg body weight as a single dose or in 3 equally divided doses within 8
hrs and observed for any abnormal symptoms of toxicity. MSCP is found to contain alkaloids (4.09 mg), phenols (0.988 mg),
flavonoids (1.223 mg) and terpenoids (2.144 mg) per gram of MSCP. The major flavonoid detected is quercetin. The MSCP is found
to possess significant radical scavenging activity against DPPH and superoxide anions (IC50 – 38.4 µg/ml) and the effect is
comparable to that of the corresponding reference compounds ascorbic acid and rutin in terms of IC50 value. By applying Karber’s
formula, 2975 mg/kg b.wt has been confirmed as the LD50value of MSCP. They found that medicinal property of C.pepo seeds may
be attributed to the presence of flavonoids and phenolic compounds which counteract the free radicals responsible for various health
complications. 6
LITERATURE REVIEW

7. 1. Rosa Martha Perez Gutierrez et. al. (2016) has under gone studies on’’ Cucurbita pepo(Pumpkin)
its Phytochemistry and Pharmacology’’. The objective of present study was to evaluate in-vitro
antioxidant and α-amylase inhibitory activity of methanolic extract of leaves. ,she undergone research
on various parts of cucurbita pepo such as fruits ,seeds etc…and studied about pharmacological
chemical constituents and clinical uses of cucurbita pepo. She identified that the extract from seed
and fruit of cucurbita pepo posses various pharmacological activity such as anti inflammatory
activity ,anti microbial activity and anti cancer/ tumour effects. The survey is mainly known for its
improvement in prostatic hyperplasia,urinary disfunction and cytotoxic properties and also been used
extensively as hypoglycemic agents.
2. Bhagyalakshmi Nair et.al .(2017) has undergone the ‘’Phytochemical screening on the seed extract
of cucurbita pepo ‘’. Which revealed the presence of Carbohydrates, Steroids, Proteins, amino acids
alkaloids, flavonoids, glycosides and tannins. They used the solutions of alcoholic and hydroalcoholic
solution and bacteria such as , Bacillus subtilis, and Escherichia coli was performed by cylindrical
plate method.Their observations reveald that Cucurbita pepo shows significant Anti microbial activity
against Escherichia coli and Bacillus subtilis activities but out of the two solutions, the hydro
alcoholic solution was giving more responses of inhibition compared to the alcoholic solution and
thus we can claim that the hydro alcoholic solution is more effective than the alcoholic solution.
7

8.  Collection of seeds of Cucurbita maxima.
 Drying and grinding of the Cucurbita maxima seeds.
 Determination of the total Ash value.
 Extraction by Soxhlet method using Hydroalcohol
solvent.
 Phytochemical screening of Hydro Alcoholic extract.
 Antioxidant activity of the extract by DPPH model.
 Antibacterial activity by Disc diffusion method.
PLAN OF WORK
8

9. The fruit of Cucurbita maxima was collected from the
tirupati market.
Seeds were obtained from the Cucurbita maxima
washed in flowing water and dried in shade .
COLLECTION OF SEEDS
9

10. MATERIALS AND METHODS
Determination of total ash value:-
5gm of Cucurbita maxima seeds powder was taken in the silica crucible
The silica crucible was placed inside the muffle furnace .
The furnace heated to 550°C and cooled .
The process was repeated till constant weight was obtained
% Ash =
W1 = Weight of empty crucible
W2=Weight of crucible with
sample
10

11. Soxhlet Method
11

12. Sl.no Constituents Test Reagents colour
1 Alkaloids Mayer’s test Mayer’s reagent Yellow colour ppt
2 Carbohydrates Molisch’s test α-naphthol solution Violet ring
3 Tanins Gelatin test 1% Gelatin solution White colour ppt
4 Glycosides Legal’s test sodium nitropruside in
pyridine and sodium
hydroxide
pink to blood red colour
5 flavonaoids lead- acetate test lead- acetate solution Yellow colour ppt
Phytochemical Screening of Hydroalcoholic Extract
12
TABLE NO 1

13. DPPH radicals scavenging activity
13
1mM solution of DPPH in ethanol.
↓
1mg/ml extract solution in ethanol was prepared.
↓
1.5ml of this solution was added to 1.5 ml of DPPH.
↓
The absorbance was measured at 517 nm against the corresponding blank solution.
↓
blank solution is prepared by taking 3ml ethanol as a control.
↓
Absorbance was determined.
X 100
Abs con - is the absorbance of the control reaction.
Abs test - is the absorbance in the presence of the sample of the extracts.

14. Nutrient agar was poured into Petri dishes and was allowed to solidify.
↓
Then bacterial culture Escherichia coli (E. coli), Staphylococcus aureus (S.aureus) is spread on the plate.
↓
Allowed it settle for 10 minutes
↓
bore or wells were prepared in each plate.
↓
1ml of dilutions of concentration 20, 60, 100μg/ml (Hydro alcoholic solutions) was poured into each plates respectively.
↓
Allow it to stand for 15 minutes.
↓
Then incubate it in incubator for 48 hours at 37 °C
.
↓
Measure the zone of inhibitions.
↓
Ciprofloxacin was taken as standard.
Antibacterial Activity
14

15. ASH VALUE
Ash value was determine with the help of the formula
and was 3.5 ,which was in the limits.
RESULTS
15

16.  Phytochemical screening was performed and the results was shown below the table
Phytochemical screening
Sl. No Constituents HAE
1 Alkaloids +
2 Carbohydrates +
3 Reducing Sugars -
4 Glycosides -
5 Saponins -
6 Triterpenes -
7 Phytosterols -
8 Phenols/Phenolic compounds. +
9 Tannins -
10 Amino Acid -
11 Diterpenes +
12 Proteins +
13 Flavanoids +
16
TABLE NO 2
Positive
Negative
+
-

17. Sl.No Constituents IC 50(µM/ml)
1 Ascorbic acid 33.9
2 H.A.E 42.4
Anti-oxidant activity
Antioxidant test was perform and the results was given below
0
5
10
15
20
25
30
35
40
45
Ascorbic acid H.A.E
IC50
Constituents
Anti Oxidant Activity Of H.A.E Extract of Seeds of Cucurbita maxima
17
TABLE NO 3
GRAPH NO 1

18. Antibacterial activity Anti-bacterial activity
of E.Coli. of S.aereus
ANTI BACTERIAL ACTIVITY
18

19. Sl.No Constituents Zone of inhibition (mm)
1. Ciprofloxacin 18
2 Control(Hydroalcohol) 6.2
3 HAE (20 µg/ml) 7.8
4 HAE (60 µg/ml) 9.4
5 HAE (100 µg/ml) 12.6
Antibacterial activity of E.Coli
0
2
4
6
8
10
12
14
16
18
20
Zoneofinhibition(mm)
Concentration
Anti Bacterial Activity of Hydro Alcoholic Extract For E.coli
19
TABLE NO 4
GRAPH NO 2

20. Sl.No Constituents Zone of inhibition (mm)
1. Ciprofloxacin 22
2 Control(Hydroalcohol) 7.2
3 HAE (20 µg/ml) 8.6
4 HAE (60 µg/ml) 12.6
5 HAE (100 µg/ml) 14.5
Antibacterial activity of HAE of seeds of Cucurbita maxima against S.aereus
0
5
10
15
20
25
Zoneofinhibition(mm)
Concentration
Anti Bacterial Activity of Hydro Alcoholic Extract For Staphylococcus
aureus
20
TABLE NO 5
GRAPH NO 3

21.  .
 All the parts of this plant has different secondary metabolites here we are collecting the
seeds of cucurbita maxima and we have extracted the constituents from dried seed powder
using hydroalcohol.
 The total ash value was determine and found with in the limits.
 The extract have shown the presence of ‘’alkaloid, flavonoids, carbohydrates, phenolic
compounds, Proteins’’.
 The anti oxidant activity using DPPH model has shown the presence of anti oxidants
property in the hydroalcoholic extract.
 The antibacterial activity against S.aureus, E.coli was carried out using cup and plate
method. The 100µg/ml concentration have shown the highest activity compared to the rest
too concentrations (20 µg/ml, 60 µg/ml).
 The standard used in the antibacterial activity was ciprofloxacin.
CONCLUSION
21

22.  SengupthaRupa, BanikJayanta,2013. Comparative Studies On
Anthelmintic Potential Of Cucurbita Maxima Seeds and Carica
Papaya Seed.Int. J. Res. Ayurveda Pharma.4(4),530-531.
 B. Abd EI-Aziz and H.H. Abd EI-Kalek,2011. Antimicrobial
proteins and oil seeds from pumpkin (Cucurbitamoschata). 9,1.
 N.Muruganantham, S Solomon, MM Senthamilselvi (2016).
Antimicrobial activity of Cucurbita maxima flowers(Pumpkin),
Journal of Pharmacognosy and Phytochemistry, 5(1).15-18.
REFERENCES
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APPENDIX

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