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detection of bioactive compounds of raphanus sativus using
gc-ms and ft-ir technical analysis and determination of
its anti-bacterial and anti-fungal activity
nebras m. sahi1, ghaidaa jihadi mohammed2, imad hadi hameed3
1department of biology, college of science for women, university of babylon, hillah city, iraq
2department of biology, college of science, university of al-qadisiyah, hillah city, iraq
3biomedical science department, university of babylon, college of nursing, hillah city, iraq
abstract
raphanus raphanistrum is an edible root vegetable of the brassicaceae family. the objective of this study
were analysis of the secondary metabolite products. twenty bioactive compounds were identified in the
methanolic extract of raphanus sativus. gc-ms analysis of raphanus sativus revealed the existence
of the raphanus sativus leaves proved the presence of alkenes, alkanes, and alkyl halides which shows
major peaks at 844.82, 937.40, 1012.63, 1022.27, 1066.64, 1143.79, 1217.08, 1234.44, 1361.74, 2330.01,
2341.58, 2358.94, and 2854.65. in the current study, the anti-microbial activity of raphanus sativus
methanolic extract was evaluated by determining the zone of inhibition against nine bacteria and eight fungi
and yeast. maximum zone formation was against staphylococcus aureus (6.75±0.25). raphanus sativus
was very highly active against aspergillus terreus (6.99±0.26).
keywords: ft-ir, gc-ms analysis, raphanus sativus, anti-bacterial, anti-fungal activity
corresponding author:
imad hadi hameed
biomedical science department,
university of babylon, college of nursing,
hillah city, iraq
phone: 009647716150716
e-mail: imad_dna@yahoo.com
introduction
radishes are grown and consumed throughout
the world, being mostly eaten raw as a crunchy salad
vegetable. radishes have long been grown as a food
crop, but they also have various medicinal actions. the
plant is used in the treatment of intestinal parasites,
though the part of the plant used is not specified 1-12. they
have numerous varieties, varying in size, flavor, color,
and length of time they take to mature. radishes owe
their sharp flavor to the various chemical compounds
produced by the plants, including glucosinolate,
myrosinase, and isothiocyanate. they are sometimes
grown as companion plants and suffer from few pests and
diseases. it is crushed and used as a poultice for burns,
bruises and smelly feet. radishes are also an excellent
food remedy for stone, gravel and scorbutic conditions
13-27. the plant contains raphanin, which is antibacterial
and antifungal. it inhibits the growth of staphylococcus\
aureus, e. coli, streptococci, pneumococci etc.radish
preparations are useful in liver and gall bladder troubles.
it is reported that the methonolic extract of leaves of
raphanus sativus active against paracetamol induced
hepato toxicity. paracetamol increced the activity of
hepatic enzymes like sgot, sgpt, serum ldh, serum
ap 28-46. an antioxidant is a molecule capable of slowing
or preventing the oxidation of other molecules. the aim
of this study were analysis of the secondary metabolite
products. the aims of our study were analysis of the
secondary metabolite products and evaluation of its
antibacterial and antifungal activity.
material and method
gas chromatography–mass spectrum analysis:
raphanus sativus gc–ms analysis were carried out
in a gc system (agilent 7890a series, usa). the flow
doi number: 10.5958/0976-5506.2018.00215.2
236 indian journal of public health research & development, march 2018, vol.9, no. 3
rate of the carrier gas, helium (he) was set to beat 1 ml
min?1, split ratio was 1:50. the injector temperature
was adjusted at 250?c, while the detector temperature
was fixed to280?c.
fourier transform infrared spectrophotometer
(ftir): the powdered sample of raphanus sativus
was treated for ftir spectroscopy (shimadzu, ir
affinity, japan). the sample was run at infrared region
between 400 nm and 4000 nm47-50.
determination of antimicrobial activity of crude
bioactive compounds of raphanus sativus: the test
pathogens were swabbed in müller-hinton agar plates.
sixty ml of plant extract was loaded on the bored wells.
antifungal activity was evaluated by measuring the zone
of inhibition against the test microorganisms. methanol
was used as solvent control51-55. amphotericin b and
fluconazole were used as reference antifungal agent.
the tests were carried out in triplicate. the antifungal
activity was evaluated by measuring the inhibition-zone
diameter observed after 48 h of incubation.
table 1: major phytochemical compounds identified in methanolic extract of raphanus sativus
molecular exact mass pharmacological actions
weight
rt
phytochemical compound (min) s.
no.
1. urea , n,n’-bis(2-hydroxyethyl)- 3.219 148 148.084792 new chemical compound
2. 2,4-pentadienenitrile 3.379 79 79.042199 antimicrobial
3. furfural 3.585 96 96.021129 anti-inflammatory
4. dimethyl trisulfide 4.306 125 125.9631634 anti-inflammation
5. propylparaben 4.781 180 180.078644 antimicrobial activity
6. 1,4-dihydrothujopsene-(l 1) 5.038 206 206.203451 new chemical compound
4-(2,5-dihydro-3-methoxyphenyl) 5.484 181 181.146665 anti-ulcer activity
7. butylamine
1-butene , 4-isothiocyanato-1- 6.137 159 159.017641 new chemical compound
8. (methylthio)-
2(3h)-naphthalenone , 4,4a,5,6,7,8- 6.211 180 180.115029 anti-inflammatory
9. hexahydro-1-methoxyantiinflammatory
10. 5-hydroxymethylfurfural 6.720 126 126.031694 properties
11. dodecanoic acid , 3-hydroxy- 7.670 216 216.1725445 antimicrobial
12. 3,4-dimethyl-5-phenyloxazolidine 9.370 177 177.115364 new chemical compound
1,7-dioxa-10-thia-4,13- 9.084 276 276.077993 new chemical compound
13. diazacyclopentadeca-5,9,12-trione
2,15-heptadecadiene , 11.269 294 294.292265 anti-termitic activity
14. 9-(ethoxymethyl)-
1,1’-(4-methyl-1,3-phenylene) 12.786 556 556.146362 anti hiv-1 activity
15. bis[3-(5-benzyl-1,3,4-thiadiazol-2
16. dexchlorpheniramine 13.020 274 274.123676 anti-allergic activity
17. olean-12-ene-3,28-diol,(3?)- 13.261 442 442.38108 anti-inflammatory
propanal , (2,4-dinitrophenyl) 14.033 238 238.070205 new chemical compound
18. hydrazone
glycine , n-[(3?,5?,12?)-3,12- 23.142 449 449.314123 new chemical compound
19. dihydroxy-24-oxocholan-24-yl]-
20. oleic acid 15.521 282 282.25588 anti-inflammatory
indian journal of public health research & development, march 2018, vol.9, no. 3 237
result s and discussion
identification of biochemical compounds: analysis
of compounds was carried out in methanolic extract of
raphanus sativus, shown in table 1. chromatogram
gc-ms analysis of the methanol extract of raphanus
sativus showed the presence of thirty one major
peaks and the components corresponding to the
peaks were determined as follows. all peaks were
determined to be urea, n,n’-bis(2-hydroxyethyl)-,
2,4-pentadienenitrile, furfural, dimethyl
trisulfide, propylparaben, 1,4-dihydrothujopsene-
(l 1), 4-(2,5-dihydro-3-methoxyphenyl)butylamine,
1-butene, 4-isothiocyanato-1-(methylthio)-,
2(3h)-naphthalenone, 4,4a,5,6,7,8-hexahydro-1-
methoxy-, 5-hydroxymethylfurfural, dodecanoic
acid, 3-hydroxy-, 3,4-dimethyl-5-phenyloxazolidine,
1,7-dioxa-10-thia-4,13-diazacyclopentadeca-5,9,12-
trione, 2,15-heptadecadiene, 9-(ethoxymethyl)-,
1,1’-(4-methyl-1,3-phenylene)bis[3-(5-benzyl-1,3,4-
thiadiazol-2, dexchlorpheniramine, olean-12-ene-3,28-
diol,(3?)-, propanal, (2,4-dinitrophenyl)hydrazine ,
dextromethorphan, oleic acid , normorphine, codeine
and glycine, n-[(3?,5?,12?)-3,12-dihydroxy-24-
oxocholan-24-yl]. the ftir analysis of raphanus
sativus leaves proved the presence of alkenes, alkanes,
and alkyl halides which shows major peaks at 844.82,
937.40, 1012.63, 1022.27, 1066.64, 1143.79, 1217.08,
1234.44, 1361.74, 2330.01, 2341.58, 2358.94, and
2854.65. in the current study, the anti-microbial
activity of raphanus sativus methanolic extract
was evaluated by determining the zone of inhibition
against nine bacteria and eight fungi and yeast. clinical
pathogens were selected for antibacterial activity
namely, staphylococcus aureus, streptococcus faecalis,
klebsiella pneumonia, salmonella typhi, salmonella
typhi, bacillus subtilis, escherichia coli, proteus
mirabilis, and streptococcus pyogenes. maximum
zone formation was against staphylococcus aureus
(6.75±0.25). methanolic extraction of raphanus
sativus showed notable antifungal activities against
penicillium expansum, aspergillus niger, aspergillus
terreus, candida albicans, saccharomyces cerevisiae,
microsporum canis, trichoderma viride, and
trichoderma horzianum. raphanus sativus was very
highly active against aspergillus terreus (6.99±0.26).
in comparison to the antibiotics used in this study, the
plants extracts were far more active against the test
bacterial strains.
conclusion
medicinal property of raphanus sativus methanolic
extract is due to presence of secondary metabolites.
twenty phytoconstituents were identified by (gc-ms)
analysis. this plant derived bioactive compounds used
as source of antibiotic properties and pharmaceutical
industries used for drug formulation.
financial disclosure: there is no financial disclosure.
conflict of interest: none to declare.
ethical clearance: in our research, all protocols were
approved under the department of biology, college of
science for women, university of babylon, hillah city,
iraq and all methods were carried out in accordance with
approved guidelines.
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  • وصف الــ Tags لهذا الموضوع
  • FT-IR, GC-MS analysis, Raphanus sativus, Anti-Bacterial, Anti-Fungal Activity

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