Determination of Antioxidant Compounds, Antibacterial Activity and Minerals Content of Broccoli - Open Journal Systems
←
→
Page content transcription
If your browser does not render page correctly, please read the page content below
Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 2799
Determination of Antioxidant Compounds, Antibacterial
Activity and Minerals Content of Broccoli
Dhuha M.A.AL-Altaie1, Zuhair Radhi Addai1
1
Scholar Researchers, Department of Biology, Faculty of Education for Pure Sciences, University of Thi-Qar, Iraq
Abstract
The aim of this study was to examine the antioxidant compounds, antibacterial activity and minerals content
of broccoli stem and floret. The antioxidant activity was determined by measuring total phenolic content
(TPC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). The antibacterial activity was determined using paper
disc method against two bacteria namely Klebsiella sp, Escherichia coli, Staphylococcus aureus and P.
aeruginosa. Minerals concentrations of broccoli stem and floret were determined using atomic absorption
spectrometry. The results showed that floret sample had significantly (P2800 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3
dietary fibre (2.60%) and carbohydrates (6.64%). It is a prepared using the same solvent employed to dissolve
rich source of minerals such as potassium, phosphorus, the broccoli extract. Zones of inhibition around the discs
calcium and sodium. The aim of research was to study were measured in mm. The experiment was repeated
the antioxidant activity and minerals content from florets in triplicate and the mean of diameter of the inhibition
and stem of broccoli. zones was calculated.
Materials and Methods Determination of minerals content
Sample collection and preparation of broccoli Extraction of minerals from broccoli stem and fruits
stem and fruits aqueous extract were performed by wet digestion according to the (13).
Dried samples were weighed in a conical flask with
The stem and fruits of broccoli were obtained from
HNO3: HCIO4 (2:1) for 3-4 hours on a sand bath at a
the market in Nasiriyah city, Iraq. The stem and fruits
temperature of 100 0C until all brown fumes had changed
of broccoli were cleaned and cut into small pieces, and
to white. Ten ml of HCL was added to the sample to
then oven dried at 60 0 C for 24 h. The dried sample was
solute the inorganic and oxide salts. Digested samples
then pulverized using a mechanical grinder and passed
were filtered with a 0.45 µm pore size cellulose nitrate
through a 250 μm mesh and then stored at 40C until use.
membrane filter paper and the volume was made up to 50
In the extraction process, approximately 1 g of Ficus
ml with deionized water. Minerals concentrations were
slurries was weighed in universal bottles and 10 ml of
determined by the Atomic absorption.
50% acetone as solvent was added.
Statistical Analysis
DPPH radical scavenging activity assay
Results are expressed as means with standard
The DPPH assay method of (11) was modified to deviation (SD) of three measurements. The significance
determine antioxidant activity using Trolox as the of differences between mean values was evaluated by
standard. For assays, 3 ml methanol DPPH solution (40 analysis of variance T-Test using SPSS vr.23 software.
mg/L) was mixed with 100 μl sample extract. Samples Differences were considered significant at a probability
were incubated in the dark at room temperature for 30 value of < 0.05.
min and then the absorbance of the solution at 517 nm
was measured. Results and Discussions
Determination of total phenolic content Antioxidant capacity assays
The total phenolic content was estimated using Antioxidant compounds react with Folin-Ciocalteu
the Folin-Ciocalteu method (11) using gallic acid as the reagent and the reaction can be performed to measure
standard. A 100-μL aliquot of plant extract was oxidized the concentration of phenolic groups (14). Therefore,
with diluted Folin-Ciocalteu reagent (500 μL). After deep blue coloration in milk samples indicates that high
5 min, the mixture was neutralized with 1 ml sodium phenolic concentrations are present, whereas light blue
carbonate (7.5%, w/v), and incubated for 120 min before coloration in milk samples indicates otherwise. The
reading absorbance at 765 nm. TPC was determined because of its strong correlation
with the antioxidant activity in various parts of broccoli
Antibacterial assay (stem and floret) (15). Based on DPPH estimations (Fig
Klebsiella sp, Escherichia coli, Staphylococcus 1 and Fig 2), floret sample had the highest percentage
aureus and P. aeruginosa were used in experiment. of antioxidant activity and stem sample had the lowest.
Mueller Hinton agar was used in antibacterial assay. There was significant difference (pIndian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 2801
the samples likewise the discrepant antioxidant values degree of discoloration indicates the scavenging potential
between the present and previous studies may be caused of an antioxidant compound. As shown in the Fig 2, the
by the differences in the vitamin C content and TPC. highest activity was in floret sample (72.08%), followed
Such differences in the results of TPC compared with by moderate activities in stem sample (53.34%). A
other researchers may be linked to different varieties comparison of the antioxidant activity values in the
of cultivars and the varying antioxidant extraction present investigation with literature data was problematic
methods used. Moreover, factors such as fruit maturity, due to the large variability and lack of standardization
agro climate and post-harvest storage conditions are of the assay methods. Therefore, the antioxidant
known to affect the content of polyphenols in fruits (16). capacities were ranked instead. The highest antioxidant
DPPH assays are often used to determine the capacity of activity was observed in different types of plant part is
primary antioxidants in samples, in which these primary an important dietary source of vitamin C, minerals and
antioxidants react to scavenge free radicals from DPPH amino acids and also contains phenolic compounds and
solution. Hence, the formation of the initiation chain of tannins. Hence, the high antioxidant activity observed
free radicals is inhibited and the propagation chain is for broccoli stem and florets in the present investigation
destroyed through the donation of a hydrogen atom or may be due to the high concentrations of vitamin C and
an electron. The bleaching of DPPH absorption by a test other antioxidant compounds. (17) have observed the
compound is representative of its capacity to scavenge highest activity using DPPH, although their antioxidant
free radicals generated independently of any enzymatic activity values are lower than those in the present study.
or transition metal-based system. This method is widely The most possible reason may be the lower quality of the
used to evaluate antioxidant activities within a relatively stem and florets used and the different maturity stages
short time compared with other methods. Antioxidants of the fruit samples. The majority of the antioxidant
react with DPPH, a stable free radical and convert it to activity of fruits is known contributed by polyphenols,
1, 1-diphenyl-2-(2, 4, 6-trinitrophenyl) hydrazine. The vitamin C, vitamin E, β-carotene and lycopene (18-19).
a–b Different letters indicate significant difference (P< 0.05)
Figure 1. Mean (n=3) Total phenolic content of broccoli stem and floret2802 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3
a–b Different
letters indicate significant difference (P< 0.05)
Figure 2. Mean (n=3) DPPH of broccoli stem and floret
Antibacterial activity broccoli samples. Based on result, the acetone extract
of (broccoli stem and floret) at a concentration of 150
The antibacterial activity of crude acetone extract and 250 μg/disc showed antibacterial activity against
of broccoli stem and floret were evaluated by using Klebsiella sp, Escherichia coli, Staphylococcus aureus
paper disc diffusion method against four types of and P. aeruginosa. Therefore, a higher concentration
bacteria namely as, Klebsiella sp, Escherichia coli, of antibacterial agent was essential to inhibit some of
Staphylococcus aureus and P. aeruginosa. Table 2 the Gram-negative bacteria. Further phenolic compound
shows the results of antibacterial activities of three types involved in adhesion binding, protein and cell wall
of broccoli samples. This work found that, antibacterial binding, enzyme inactivation, and intercalation into the
had significant different (pIndian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 2803
Minerals content the stem and floret of broccoli. However, Sodium,
Calcium and Magnesium are the most abundant elements
Since minerals are the big concerns for the public
in both broccoli samples. In present study broccoli floret
health today, so this study carried out to measure the
samples have been found to contain the highest value of
contents of minerals in broccoli stem and floret by
K, Ca, Mg, Mn, Cu, Fe and Zn (2.19, 1245.43, 364.67,
using Atomic Absorption Spectrometry. Fourteen
20.09, 2.18, 20.72 and 6.63 mg/100g DW respectively),
minerals were analyzed in this study namely, Sodium
whereas the lowest amount was found in broccoli stem
(Na), Potassium (K), Calcium (Ca), Magnesium (Mg),
samples (1.41, 468.33, 350.00, 10.41, 1.29, 10,49
Manganese (Mn), Copper (Cu), Iron (Fe), Zinc (Zn).
and 4.95 mg/100g DW respectively). The results are
Table 2. illustrates minerals content of two samples of
consistent with those obtained from the study conducted
broccoli plant. Based on the results, it was observed that,
by (21-22) who evaluated the quantity of Calcium (Ca),
Sodium (Na), Potassium (K), Calcium (Ca), Magnesium
Sodium (Na), Magnesium (Mg), Potassium (K), Iron
(Mg), Manganese (Mn), Copper (Cu), Iron (Fe), Zinc
(Fe) and Copper (Cu) of broccoli samples from different
(Zn). Showed significant difference (p2804 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3
3- Grover JK, Yadav S, and Vats V. Medicinal plants In Press, http://dx.doi.org/10.1016/j. food chem.
of India with anti-diabetic potential. 2002. 2014.03.039.
4- Aires, A., Fernandes, C., Carvalho, R., Bennett, 13- AOAC, American Society of Analytical Chemistry
R.N., Saavedra, M.J., and Rosa, E.A.S. Seasonal and Preparation Method. 2000. 999:10.
effects on bioactive compounds and antioxidant 14- Nurliyana, R., I. Syed Zahir, K. Mustapha
capacity of six economically important Brassica Suleiman, M.R. Aisyah and K. Kamarul Rahim,
vegetables. Molecules 2011. 16:6816-6832. Antioxidant study of pulps and peels of dragon
5- Bhandari, S.R., and Kwak, J.H. Chemical fruits: a comparative study. Int. Food Res. J., 2010.
composition and antioxidant activity in different 17: 367-375.
tissues of Brassica vegetables. Molecules 2015. 15- Gorinstein, S., Z. Zachwieja, E. Katrich, E.
20:1228-1243. Pawelzik, R. Haruenkit, S. Trakhtenberg and O.
6- Jo, J.S., Bhandari, S.R., Kang, G.H., and Lee, J.G. Martin-Belloso, Comparison of the contents of the
Comparative analysis of individual glucosinolates, main antioxidant compounds and the antioxidant
phytochemicals, and antioxidant activities in activity of white grapefruit and his new hybrid.
broccoli breeding lines. Horticulture Environment LWT-Food Sci. Technol., 2004. 37(3): 337-343.
and Biotechnology 2016. 57(4):392-403. 16- Mahattanatawee, K., J.A. Manthey, G. Luzio,
7- Razis, A.F.A., and Noor, N.M. Cruciferous S.T. Talcott, K. Goodner and E.A. Baldwin, Total
vegetables: dietary phytochemicals for cancer antioxidant activity and fiber content of select
prevention. Asian Pacific Journal of Cancer Florida-grown tropical fruits. J. Agric. Food Chem.,
Prevention 2013. 14 (3):1565-1570. 2006. 54(19): 7355-7363.
8- Bachiega, P., Salgado, J.M., De Carvalho, J.E., 17- Leong, L.P. and G. Shui, An investigation of
Ruiz, A.L.T.G., Schwarz, K., Tezotto, T., et al. antioxidant capacity of fruits in Singapore markets.
Antioxidant and antiproliferative activities in Food Chem., 2002. 76(1): 69-75.
different maturation stages of broccoli (Brassica 18- Gancel, A.L., P. Alter, C. Dhuique-Mayer, J.
oleracea Italica) biofortified with selenium. Food Ruales and F. Vaillant, Identifying carotenoids
Chemistry 2016. 190:771-776. and phenolic compounds in naranjilla (Solanum
9- Podsedek, A. Natural antioxidants and antioxidant quitoense Lam. var. Puyo hybrid), an andean
capacity of Brassica vegetables: a review. LWT- fruit. J. Agric. Food Chem., 56(24): 11890-11899.
Food Science and Technology 2007. 40:1-11. Madhu and Anita Kochhar. 2014. Proximate
10- Olga N, Campas-Baypoli, Dalia I Sa´Nchez- composition, available carbohydrates, dietary fibre
Machado, Carolina Bueno-Solano, Jose A, Nu´ N˜ and anti-nutritional factors of Broccoli (Brassica
Ez-Gaste´Lum, Cuauhte´Moc Reyes-Moreno and oleracea L. Var. Italica Plenck) leaf and floret
Jaime Lo´ Pez-Cervantes, Biochemical composition powder. Bioscience Discovery, 2008. 5(1):45-49.
and physicochemical properties of broccoli flours. 19- Beatriz de la Fuente , Gabriel López-García, Vicent
Int. J. Food Sci. and Nutr, 2009. 60(S4): 163-173. Máñez, Amparo Alegría, Reyes Barberá and
11- Musa, K. H., Abdullah, A., Jusoh, K. & Antonio Cilla. Evaluation of the Bioaccessibility
Subramaniam, V. Antioxidant activity of pink-flesh of Antioxidant Bioactive Compounds and Minerals
guava (Psidium guajava L.): effect of extraction of Four Genotypes of Brassicaceae Microgreens.
techniques and solvents. Food Analytical Methods Foods, 2019. 8, 250: 1-16.
2011. 4: 100-107. 20- Pereira J.A, Oliveira I, Sousa A, Valento P, Andrade
12- Kumar V.N, Murthy P.S, Manjunatha J.R. and P.B, FerreiraI.C.F.R, Ferreres F, Bento A, Seabra
Bettadaiah B.K, Synthesis and quorum sensing R. and Estevinho L. Walnut (Juglansregia L.)
inhibitory activity of key phenolic compounds of leaves: phenolic compounds, antimicrobial activity
ginger and their derivatives, Food Chem., 2014, and antioxidant potential of different cultivars,
Food Chem. 2007. 45: 2287–2295.You can also read
