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.
references
1. gilani ah, janbaz kh, akhtar ms. selective
protective effect of an extract from fumaria
parviflora on paracetamol induced hepatotoxicity.
gen. pharmacol. 1996 27: 979-983
2. mohammed gj, kadhim mj, hameed ih.
proteus species: characterization and herbal
antibacterial: a review. international journal of
pharmacognosy and phytochemical research.
2016 8(11): 1844-1854.
3. shireen sk, hameed ih, hamza lf. acorus
calamus: parts used, insecticidal, anti-fungal,
antitumour and anti-inflammatory activity: a
review. international journal of pharmaceutical
quality assurance. 2017 8(3): 153-157.
4. huda ja, hameed ih, hamza lf. anethum
graveolens: physicochemical properties,
medicinal uses, antimicrobial effects, antioxidant
effect, anti-inflammatory and analgesic effects: a
review. international journal of pharmaceutical
quality assurance. 2017 8(3): 88-91.
5. altaee n, kadhim mj, hameed ih. detection of
volatile compounds produced by pseudomonas
aeruginosa isolated from uti patients by gas
chromatography-mass spectrometry. international
journal of toxicological and pharmacological
research. 2016 8(6): 462-470.
238 indian journal of public health research & development, march 2018, vol.9, no. 3
6. hussein hm, hameed ih, ubaid jm. analysis of
the secondary metabolite products of ammi majus
and evaluation anti-insect activity. international
journal of pharmacognosy and phytochemical
research. 2016 8(8): 1192-1189.
7. hussein hm, ubaid jm, hameed ih. inscticidal
activity of methanolic seeds extract of ricinus
communis on adult of callosobruchus maculatus
(coleopteran:brauchidae) and analysis of its
phytochemical composition. international journal
of pharmacognosy and phytochemical research.
2016 8(8): 1385-1397.
8. ubaid jm, hussein hm, hameed ih.
determination of bioactive chemical composition
of callosobruchus maculutus and investigation
of its anti-fungal activity. international journal
of pharmcognosy and phytochemical research.
2016 8(8): 1293-1299.
9. ibraheam ia, hussein hm, hameed ih.
cyclamen persicum: methanolic extract using
gas chromatography-mass spectrometry
(gc-ms) technique. international journal of
pharmaceutical quality assurance. 2017 8(4)
200-213.
10. ibraheam ia, hadi my, hameed ih. analysis
of bioactive compounds of methanolic
leaves extract of mentha pulegium using
gas chromatography-mass spectrometry
(gc-ms) technique. international journal of
pharmaceutical quality assurance. 2017 8(4)
174-182.
11. hadi my, hameed ih, ibraheam ia. ceratonia
siliqua: characterization, pharmaceutical
products and analysis of bioactive compounds:
a review. research journal of pharmacy and
technology. 2017 10(10): 3585-3589.
12. hadi my, hameed ih, ibraheam ia. mentha
pulegium: medicinal uses, anti-hepatic,
antibacterial, antioxidant effect and analysis
of bioactive natural compounds: a review.
research journal of pharmacy and technology.
2017 10(10): 3580-3584.
13. kadhim mj, sosa aa, hameed ih. evaluation
of anti-bacterial activity and bioactive
chemical analysis of ocimum basilicum using
fourier transform infrared (ft-ir) and gas
chromatography-mass spectrometry (gcms)
techniques. international journal of
pharmacognosy and phytochemical research.
2016 8(6): 127-146.
14. mohammed gj, kadhim mj, hussein hm.
characterization of bioactive chemical compounds
from aspergillus terreus and evaluation of
antibacterial and antifungal activity. international
journal of pharmacognosy and phytochemical
research. 2016 8(6): 889-905.
15. hameed ih, altameme hj, idan sa. artemisia
annua: biochemical products analysis of
methanolic aerial parts extract and anti-microbial
capacity. research journal of pharmaceutical,
biological and chemical sciences. 2016 7(2):
1843- 1868
16. mohammed gj, omran am, hussein hm.
antibacterial and phytochemical analysis of
piper nigrum using gas chromatography-
mass spectrum and fourier-transform
infrared spectroscopy. international journal of
pharmacognosy and phytochemical research.
2016 8(6): 977-996.
17. jasim h, hussein ao, hameed ih, kareem
ma. characterization of alkaloid constitution
and evaluation of antimicrobial activity of
solanum nigrum using gas chromatography mass
spectrometry (gc-ms). journal of pharmacognosy
and phytotherapy. 2015 7(4): 56-72.
18. hadi my, mohammed gj, hameed ih. analysis
of bioactive chemical compounds of nigella sativa
using gas chromatography-mass spectrometry.
journal of pharmacognosy and phytotherapy.
2016 8(2): 8-24.
19. hameed ih, ibraheam ia, kadhim hj. gas
chromatography mass spectrum and fouriertransform
infrared spectroscopy analysis of
methanolic extract of rosmarinus oficinalis leaves.
journal of pharmacognosy and phytotherapy.
2015 7 (6): 90-106.
20. shareef hk, muhammed hj, hussein hm,
hameed ih. antibacterial effect of ginger
(zingiber officinale) roscoe and bioactive
chemical analysis using gas chromatography
mass spectrum. oriental journal of chemistry.
2016 32(2): 20-40.
21. al-jassaci mj, mohammed gj, hameed
ih. secondary metabolites analysis of
saccharomyces cerievisiae and evaluation of
antibacterial activity. international journal of
indian journal of public health research & development, march 2018, vol.9, no. 3 239
pharmaceutical and clinical research. 2016
8(5): 304-315.
22. mohammed gj, al-jassani mj, hameed ih.
anti-bacterial, antifungal activity and chemical
analysis of punica grantanum (pomegranate
peel) using gc-ms and ftir spectroscopy.
international journal of pharmacognosy and
phytochemical research. 2016 8(3): 480-494.
23. dhahir bm, hameed ih, jaber ar. prospective
and retrospective study of fractures according
to trauma mechanism and type of bone fracture.
research journal of pharmacy and technology.
2017 10(10):1827-1835.
24. hapeep ma, hameed ih, jasim aa. risk factors,
cause and site of firearm injuries: a prospective and
retrospective study. research journal of pharmacy
and technology. 2017 10(10): 3420-3425.
25. jasim aa, hameed ih, hapeep ma. traumatic
events in an urban and rural population of
children, adolescents and adults in babylon
governorate - iraq. research journal of pharmacy
and technology. 2017 10(10): 3429-3434.
26. altameme hj, hameed ih, abu-serag na.
analysis of bioactive phytochemical compounds
of two medicinal plants, equisetum arvense
and alchemila valgaris seed using gas
chromatography-mass spectrometry and fouriertransform
infrared spectroscopy. malays. appl.
biol. 2015 44(4): 47–58.
27. hussein hm, hameed ih, ibraheem oa.
antimicrobial activity and spectral chemical
analysis of methanolic leaves extract of
adiantum capillus-veneris using gc-ms and
ft-ir spectroscopy. international journal of
pharmacognosy and phytochemical research.
2016 8(3): 369-385.
28. kadhim mj, mohammed gj, hameed ih. in
vitro antibacterial, antifungal and phytochemical
analysis of methanolic fruit extract of cassia
fistula. oriental journal of chemistry. 2016
32(2): 10-30.
29. hussein hm. analysis of trace heavy metals
and volatile chemical compounds of lepidium
sativum using atomic absorption spectroscopy,
gas chromatography-mass spectrometric and
fourier-transform infrared spectroscopy. research
journal of pharmaceutical, biological and
chemical sciences. 2016 7(4): 2529 – 2555.
30. jaddoa hh, hameed ih, mohammed gj. analysis
of volatile metabolites released by staphylococcus
aureus using gas chromatography-mass
spectrometry and determination of its antifungal
activity. oriental journal of chemistry. 2016
32(4): 8-24.
31. hameed ih, salman hd, mohammed gj.
evaluation of antifungal and antibacterial
activity and analysis of bioactive phytochemical
compounds of cinnamomum zeylanicum
(cinnamon bark) using gas chromatography-mass
spectrometry. oriental journal of chemistry.
2016 32(4): 16-25.
32. kadhim mj, mohammed gj, hussein hm.
analysis of bioactive metabolites from candida
albicans using (gc-ms) and evaluation of
antibacterial activity. international journal of
pharmaceutical and clinical research. 2016
8(7): 655-670.
33. ubaid jm, hussein hm, hameed ih. analysis of
bioactive compounds of tribolium castaneum and
evaluation of anti-bacterial activity. international
journal of pharmaceutical and clinical research.
2016 8(7): 655-670.
34. hameed, i.h., al-rubaye a.f. and kadhim, m.j.
antimicrobial activity of medicinal plants and
urinary tract infections. international journal
of pharmaceutical and clinical research. 2017
8(11): 44-54.
35. kadhim wa, kadhim, m.j., hameed, i.h.
antibacterial activity of several plant extracts
against proteus species. international journal
of pharmaceutical and clinical research. 2017
8(11): 88-94.
36. ahmed md, hameed ih, abd-ali mq.
prospective and retrospective study of the acute
heart attack cases in marjan hospital-hillah
city-iraq. research journal of pharmacy and
technology. 2017 10(10): 3408-3416.
37. fakhir df, hameed ih, flayyih ss. burns
injuries: a prospective statistical study of 112
patients. research journal of pharmacy and
technology. 2017 10(10): 3401-3407.
38. mekhlef ak, hameed ih, khudhair me.
prevalence of physical injuries on the head, neck
and entire body in, hilla, iraq. research journal
of pharmacy and technology. 2017 10(10):
3276-3282.
240 indian journal of public health research & development, march 2018, vol.9, no. 3
39. hameed ih, al-rubaye af, kadhim mj.
antimicrobial activity of medicinal plants and
urinary tract infections. international journal
of pharmaceutical and clinical research. 2017
9(1): 44-50.
40. al-rubaye af, hameed ih, kadhim mj. a
review: uses of gas chromatography-mass
spectrometry (gc-ms) technique for analysis
of bioactive natural compounds of some plants.
international journal of toxicological and
pharmacological research. 2017 9(1) 81-85.
41. kadhim mj, kaizal af, hameed ih. medicinal
plants used for treatment of rheumatoid
arthritis: a review. international journal of
pharmaceutical and clinical research. 2016
8(12): 1685-1694.
42. ubaid jm, kadhim mj, hameed ih. study of
bioactive methanolic extract of camponotus fellah
using gas chromatography – mass spectrum.
international journal of toxicological and
pharmacological research. 2016 8(6): 434-439.
43. hussein hm, hameed ih, ubaid jm. analysis of
the secondary metabolite products of ammi majus
and evaluation anti-insect activity. international
journal of pharmacognosy and phytochemical
research. 2016 8(8): 1192-1189.
44. hussein hm, ubaid jm, hameed ih. inscticidal
activity of methanolic seeds extract of ricinus
communis on adult of callosobruchus maculatus
(coleopteran:brauchidae) and analysis of its
phytochemical composition. international journal
of pharmacognosy and phytochemical research.
2016 8(8): 1385-1397.
45. ubaid jm, hussein hm, hameed ih.
determination of bioactive chemical composition
of callosobruchus maculutus and investigation
of its anti-fungal activity. international journal
of pharmcognosy and phytochemical research.
2016 8(8): 1293-1299.
46. hussein jh, hameed ih, hadi my. using gas
chromatography-mass spectrometry (gc-ms)
technique for analysis of bioactive compounds
of methanolic leaves extract of lepidium sativum.
research journal of pharmacy and technology.
2017 10 (11): 3981-3989.
47. hadi my, hameed ih. uses of gas
chromatography-mass spectrometry (gc-ms)
technique for analysis of bioactive chemical
compounds of lepidium sativum: a review.
research journal of pharmacy and technology.
2017 10 (11): 4039-4042.
48. ubaid jm, hadi my, hameed ih. bioactive
chemical compounds identified in methanolic
extract of trogoderma granarium. research
journal of pharmacy and technology. 2017 10
(11): 3997-4004.
49. hameed ih, calixto mr, hadi my. antimicrobial,
antioxidant, hemolytic, anti-anxiety, and
antihypertensive activity of passiflora species.
research journal of pharmacy and technology.
2017 10 (11): 4079-4084.
50. hameed ih, calixto mr, hadi my. a review:
solanum nigrum l. antimicrobial, antioxidant
properties, hepatoprotective effects and analysis
of bioactive natural compounds. research
journal of pharmacy and technology. 2017 10
(11): 4063-4068.
51. hussein jh, hameed ih, hadi my. a review:
anti-microbial, anti-inflammatory effect and
cardiovascular effects of garlic: allium sativum.
research journal of pharmacy and technology.
2017 10 (11): 4069-4078.
52. flayyih ss, hameed ih, fakhir fd. road traffic
accident coming to hillah teaching hospital:
prospective and retrospective study. research
journal of pharmacy and technology. 2017 10
(11): 3819-3825.
53. fakhir df, hameed ih, flayyih ss. retrospective
study: burn injury from 2010 to 2015 in a burn
unit-hillah teaching hospital-iraq. research
journal of pharmacy and technology. 2017 10
(11): 3831-3838.
54. khudhair me, hameed ih, mekhlef ak. a
prospective and retrospective study of acute
bronchitis in hillah city-iraq. research journal
of pharmacy and technology. 2017 10 (11):
3839-3844.
55. kamal sa, hamza lf, ibraheam ia.
characterization of antifungal metabolites
produced by aeromonas hydropinghila and analysis
of its chemical compounds using gc-ms.
research journal of pharmacy and technology.
2017 10 (11): 3845-3851.