Analysis of secondary metabolites released by pseudomonas fluorescens using gc ms technique and determination of its anti-fungal activity

1. Analysis of Secondary Metabolites Released by Pseudomonas
fluorescens Using GC-MS Technique and Determination of Its
Anti-Fungal Activity
Rafid Hadi Hameed1
, Fatima Moeen Abbas2
, Imad Hadi Hameed3
1
Ministry of Public Health, Maysan Health Department, Mesan governorate, Iraq, 2
Department of Biology,
College of Science for Women, University of Babylon, Hillah City, Iraq, 3
Biomedical Science Department,
University of Babylon, College of Nursing, Hillah City, Iraq
ABSTRACT
Pseudomonas fluorescens is a common gram-negative, rod-shaped bacterium [1]. It belongs to the
Pseudomonas genus. The objectives of this study were analysis of the secondary metabolite products and
evaluation antimicrobial activity. Twenty four bioactive compounds were identified in the methanolic
extract of Pseudomonas fluorescens. GC-MS analysis of Pseudomonas fluorescens revealed the existence
of the cis-5,8,11,14,17-Eicosapentaenoic acid, 12,15-Octadecadiynoic acid, methyl ester, 7-epi-cis-
sesquisabinene hydrate, α-D-Glucopyranoside , O-α-D-glucopyranosyl, Acetamide, N-methyl-N-[4-
[2-acetoxymethyl-1- pyrrol, Acetamide, N-methyl-N-[4-[2-fluoromethyl-1- pyrrolid, Phen-1,4-diol ,
2,3-dimethyl-5-trifluoromethyl, Geranyl isovalerate, Quinazoline, 4-methyl, Pentetic acid, trans-13-
Octadecenoic acid, 9-Hexadecenoic acid, 13-Hexyloxacyclotridecan -2-one, 7-Methyl-Z-tetradecen-1- ol
acetate, cis-13-Eicosenoic acid, Didemin B, Hexadecanoic acid ,1-(hydroxymethyl) -1,2-ethaned, and Ethyl
iso –allocholate. Cassia angustifolia (Crude) was very highly active (6.007±0.20) mm against Pseudomonas
fluorescens. The results of anti-fungal activity produced by Pseudomonas fluorescens showed that the
volatile compounds were highly effective to suppress the growth of Trichoderma horzianum (5.019±0.18).
Pseudomonas fluorescens produce many important secondary metabolites with high biological activities.
Keywords: Anti-Microbial, Pseudomonas fluorescens, GC-MS, Secondary metabolites.
Corresponding author:
Imad Hadi Hameed
Biomedical Science Department, University of
Babylon, College of Nursing, Hillah city, Iraq;
Phone number: 009647716150716;
E-mail: imad_dna@yahoo.com
INTRODUCTION
P. fluorescens has multiple flagella. It has an
extremely versatile metabolism, and can be found in the
soil and in water. It is an obligate aerobe, but certain
strains are capable of using nitrate instead of oxygen as
a final electron acceptor during cellular respiration. P.
fluorescens is an unusual cause of disease in humans,
and usually affects patients with compromised immune
systems (e.g., patients on cancer treatment) 1-6
. P.
fluorescens isolates produce the secondary metabolite
2,4-diacetylphloroglucinol (2,4-DAPG), the compound
found to be responsible for antiphytopathogenic and
biocontrol properties in these strains. Several strains of P.
fluorescens, such as Pf-5 and JL3985, have developed a
natural resistance to ampicillin and streptomycin. These
antibiotics are regularly used in biological research as a
selective pressure tool to promote plasmid expression7-11
.
The aims of our research were analysis of the secondary
metabolite products of Pseudomonas fluorescens and
evaluation antimicrobial activity.
MATERIALS AND METHOD
Detection of secondary metabolites
Subcultures were obtained on the nutrient agar for 48
hrs. at 22Cº. Metabolites was separated from the liquid
cultureandevaporatedtodrynesswitharotaryevaporator
DOI Number: 10.5958/0976-5506.2018.00485.0

2. 446 Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5
at 45Cº. The residue was dissolved in 1 ml methanol,
filtered through a 0.2 μm syringe filter, and stored at 4Cº
for 24 h before being used for gas chromatography mass
spectrometry 12-23
. The identification of the components
was based on comparison of their mass spectra with those
of NIST mass spectral library as well as on comparison
of their retention indices either with those of authentic
compounds or with literature values24-35
.
Spectral analysis of bioactive natural chemical
compounds of Pseudomonas fluorescens using (GC/
MS)
Analysis was conducted using GC-MS (Agilent 789
A) equipped with a DB-5MS column (30 m×0.25 mm
i.d., 0.25 um film thickness, J&W Scientific, Folsom,
CA). The oven temperature was programmed as for the
previous analysis36-39
. Helium was used as the carrier gas
at the rate of 1.0 mL/min. Effluent of the GC column
was introduced directly into the source of the MS via a
transfer line (250 Cº)40-42
.
Determination of antibacterial and antifungal
activity
Five-millimeter diameter wells were cut from the
agar using a sterile cork-borer, and 25 μl of the plant
samples solutions were delivered into the wells. The
plates were incubated for 48 h at room temperature.
Antimicrobial activity was evaluated by measuring the
zone of inhibition against the test microorganisms43-47
.
The studied fungi, Microsporum canis, Aspergillus
flavus, Candida albicans, S. cerevisiae, Trichoderma
viride, Trichoderma horzianum, and Aspergillus terreus
were isolated and maintained in potato dextrose agar
slants. Spores were grown in a liquid culture of potato
dextrose broth (PDB) and incubated at 25ºC in a shaker
for 16 days at 130 rpm. The extraction was performed
by adding 25 ml methanol to 100 ml liquid culture in
an Erlenmeyer flask after the infiltration of the culture.
Methanol was used as solvent control. Amphotericin
B and fluconazole were used as reference antifungal
agent 48-50
. Results of the study were based on analysis
of variance (ANOVA) using Statistica Software. A
significance level of 0.05 was used for all statistical tests.
RESULTS AND DISCUSSION
Gas chromatography and mass spectroscopy
analysis of compounds was carried out in methanolic
extract of Pseudomonas fluorescens, shown in Table 1.
GC-MS analysis of Pseudomonas fluorescens revealed
the existence of the cis-5,8,11,14,17-Eicosapentaenoic
acid, 12,15-Octadecadiynoic acid , methyl ester, 7-epi-
cis-sesquisabinene hydrate, α-D-Glucopyranoside ,
O-α-D-glucopyranosyl, Acetamide , N-methyl-N-[4-[2-
acetoxymethyl-1-pyrrol,Acetamide, N-methyl-N-[4-[2-
fluoromethyl-1- pyrrolid, Phen-1,4-diol , 2,3-dimethyl-
5-trifluoromethyl, Geranyl isovalerate, Quinazoline ,
4-methyl, Pentetic acid, trans-13-Octadecenoic acid,
9-Hexadecenoic acid, 13-Hexyloxacyclotridecan
-2-one, 7-Methyl-Z-tetradecen-1- ol acetate, cis-
13-Eicosenoic acid, Didemin B, Hexadecanoic acid
,1-(hydroxymethyl) -1,2-ethaned, and Ethyl iso –
allocholate. The results of anti-fungal activity produced
by Pseudomonas fluorescens showed that the volatile
compounds were highly effective to suppress the
growth of Trichoderma horzianum (5.019±0.18)
Table 2. Pseudomonas fluorescens produce many
important secondary metabolites with high biological
activities. Based on the significance of employing
bioactive compounds in pharmacy to produce drugs
for the treatment of many diseases, the purification of
compounds produced by Pseudomonas fluorescens can
be useful. In agar well diffusion method the selected
medicinal plants Nerium olender (Alkaloids), Ricinus
communis (Alkaloids), Datura stramonium(Alkaloids),
Linum usitatissimum (Crude), Anastatica hierochuntica
(Crude), Cassia angustifolia (Crude), Althaea rosea
(Crude), Coriandrum sativum (Crude), Origanum
vulgare (Crude), Urtica dioica (Crude), Foeniculum
vulgare (Crude), and Ocimum basilicum (Crude)
were effective against Staphylococcus aureus, Table
3. Cassia angustifolia (Crude) was very highly active
(6.007±0.20) mm against Pseudomonas fluorescens.
Pseudomonas fluorescens was found to be sensitive to
all test medicinal plants and mostly comparable to the
standard reference antifungal drug Amphotericin B and
fluconazole to some extent.

3. Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5 447
Table 1. Major chemical compounds identified in methanolic extract of Pseudomonas fluorescens.
Molecular WeightRT (min)Phytochemical compoundSerial No.
302.224583.184cis-5,8,11,14,17-Eicosapentaenoic acid1.
290.224583.29312,15-Octadecadiynoic acid , methyl ester2.
222.1983653.6597-epi-cis-sesquisabinene hydrate3.
504.1690354.283α-D-Glucopyranoside , O-α-D-glucopyranosyl4.
266.1630424.609Acetamide , N-methyl-N-[4-[2-acetoxymethyl-1- pyrrol5.
226.1481424.672Acetamide , N-methyl-N-[4-[2-fluoromethyl-1- pyrrolid6.
206.0554645.295Phen-1,4-diol , 2,3-dimethyl-5-trifluoromethyl-7.
238.193286.016Geranyl isovalerate8.
144.0687487.899Quinazoline , 4-methyl-9.
393.1383448.322Pentetic acid10.
282.255888.551trans-13-Octadecenoic acid11.
254.2245810.7489-Hexadecenoic acid12.
282.2558812.22513-Hexyloxacyclotridecan -2-one13.
268.2402313.3187-Methyl-Z-tetradecen-1- ol acetate14.
310.2871813.375cis-13-Eicosenoic acid15.
308.1987613.6781,9-Dioxacyclohexadeca - 4,13-diene-2-10-dione , 716.
322.32356613.701Z-5-Methyl-6-heneicosen-11-one17.
1111.6416614.525Didemin B18.
568.50667614.719Hexadecanoic acid ,1-(hydroxymethyl) -1,2-ethaned19.
404.26750818.130Pregn-4-ene-3,20-dione , 17,21-dihydroxy-, bis(O-me20.
286.1932818.679Androst -5,7-dien-3-ol-17-one21.
488.24101819.743(22S)-21-Acetoxy-6α , 11β-dihydroxy-16α 17α-propy22.
436.31887421.843Ethyl iso -allocholate23.
422.19406722.7594H-Cyclopropa[5’,6’]bens[1’,2’:7,8]azuleno[5,6-b]ox24.
Table 2. Antifungal activity of Pseudomonas fluorescens metabolite products.
Microorganism Pseudomonas fluorescens products Fluconazol
Microsporum canis 3.116±0.16 ª 3.110±0.15
Aspergillus flavus 5.000±0.19 4.309±0.17
Candida albicans 4.702±0.17 2.873.±0.12
S. cerevisiae 3.005±0.16 2.000±0.11
Trichoderma viride 4.957±0.17 1.704±0.10
Trichoderma horzianum 5.019±0.18 4.005±0.19
Aspergillus terreus 4.951±0.16 3.251±0.17
ª The values ( average of triplicate) are diameter of zone of inhibition at 100 mg/mL crude extract and 30 μg/mL
of (Amphotericin B; Fluconazol and Miconazole nitrate).

4. 448 Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5
Table 3. Zone of inhibition (mm) of test different
bioactive compounds and standard antibiotics of
medicinal plants to Pseudomonas fluorescens.
S.
No.
Plant
Zone of
inhibition
(mm)
1. Nerium olender (Alkaloids) 3.779±0.17
2. Ricinus communis (Alkaloids) 2.639±0.15
3. Datura stramonium(Alkaloids) 3.990±0.16
4. Linum usitatissimum (Crude) 4.815±0.18
5. Anastatica hierochuntica (Crude) 5.941±0.19
6. Cassia angustifolia (Crude) 6.007±0.20
7. Althaea rosea (Crude) 5.000±0.18
8. Coriandrum sativum (Crude) 5.943±0.19
9. Origanum vulgare (Crude) 5.799±0.18
10. Urtica dioica (Crude) 4.228±0.16
11. Foeniculum vulgare (Crude) 3.000±0.14
12. Ocimum basilicum (Crude) 5.716±0.17
13. Control 0.00
CONCLUSION
Twenty four bioactive chemical constituents
have been identified from methanolic extract of the
Pseudomonas fluorescens by GC-MS. In vitro antifungal
and antibacterial evaluation of secondary metabolite
products of Pseudomonas fluorescens forms a primary
platform for further phytochemical and pharmacological
investigation for the development of new potential
antimicrobial compounds.
Financial disclosure: There is no financial
disclosure.
Conflict of Interest: None to declare.
Ethical Clearance: All experimental protocols
were approved under the Department of Biology, College
of Science, Hillah city, Iraq and all experiments were
carried out in accordance with approved guidelines.
REFERENCES
1. 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.
2. 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.
3. 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.
4. 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.
5. Ubaid JM, Hussein HM, Hameed IH. Determination
ofbioactivechemicalcompositionofCallosobruchus
maculutus and investigation of its anti-fungal
activity. International journal of pharmcognosy and
phytochemical research. 2016; 8(8): 1293-1299.
6. 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.
7. 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.
8. Gilani AH, Janbaz KH, Akhtar MS. Selective
protective effect of an extract from fumaria
parviflora on paracetamol induced hepato-toxicity.
Gen. pharmacol. 1996; 27: 979-983
9. 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.
10. 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.

5. Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5 449
11. 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.
12. Kadhim MJ, Sosa AA, Hameed IH. Evaluation of
anti-bacterialactivityandbioactivechemicalanalysis
of Ocimum basilicum using Fourier transform
infrared (FT-IR) and gas chromatography-mass
spectrometry (GC-MS) techniques. International
Journal of Pharmacognosy and Phytochemical
Research. 2016; 8(6): 127-146.
13. 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.
14. 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
15. 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.
16. 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.
17. 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.
18. Al-Jassaci MJ, Mohammed GJ, Hameed IH.
Secondary Metabolites Analysis of Saccharomyces
cerievisiae and Evaluation of Antibacterial Activity.
International Journal of Pharmaceutical and Clinical
Research. 2016; 8(5): 304-315.
19. Mohammed GJ, Al-Jassani MJ, Hameed IH. Anti-
bacterial,AntifungalActivity 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.
20. 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.
21. 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.
22. 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.
23. 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 fourier-transform infrared
spectroscopy. Malays. Appl. Biol. 2015; 44(4):
47–58.
24. Hussein HM, Hameed IH, Ibraheem OA.
AntimicrobialActivityandspectralchemicalanalysis
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.
25. 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.
26. 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.
27. Hameed IH, SalmanHD, 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

6. 450 Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5
Journal of Chemistry. 2016; 32(4): 16-25.
28. 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.
29. 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.
30. 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.
31. 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.
32. 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.
33. 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.
34. MekhlefAK, 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.
35. 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.
36. 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.
37. 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.
38. 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.
39. Ubaid JM, Hussein HM, Hameed IH. Determination
ofbioactivechemicalcompositionofCallosobruchus
maculutus and investigation of its anti-fungal
activity. International journal of pharmcognosy and
phytochemical research. 2016; 8(8): 1293-1299.
40. 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.
41. HadiMY,HameedIH.UsesofGasChromatography-
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.
42. 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.
43. 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.
44. 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.
45. KamalSA,HamzaLF,IbraheamIA.Characterization
of Antifungal Metabolites Produced by Aeromonas
hydrophila andAnalysis of its Chemical Compounds
Using GC-MS. Research Journal of Pharmacy and
Technology. 2017; 10 (11): 3845-3851.
46. Sahi NM, Mohammed GJ, Hameed IH. Detection

7. Indian Journal of Public Health Research & Development, May 2018, Vol. 9, No. 5 451
of Bioactive Compounds of Raphanus sativus
Using GC-MS and FT-IR Technical Analysis and
Determination of its Anti-Bacterial and Anti-Fungal
Activity. Indian Journal of Public Health Research
and Development. 2018; 9(3): 235-240.
47. Hameed RH, Mohammed GJ, Hameed IH.
Characterization of Antimicrobial Metabolites
Produced by Salvadora persica and Analysis of Its
Chemical Compounds Using GC-MS and FTIR.
Indian Journal of Public Health Research and
Development. 2018; 9(3): 241-246.
48. Mohammed GJ, Hameed IH, Kamal SA.
Determination of Bioactive chemical Compounds
of Aspergillus flavus Using GC/MS and FTIR and
Evaluation of Its Anti-Microbial activity. Indian
Journal of Public Health Research and Development.
2018; 9(3): 247-253.
49. Kamal SA, Mohammed GJ, Hameed IH.
Antimicrobial, Anti-inflammatory, Analgesic
Potential and Cytotoxic Activity of Salvadora
persica: A review. Indian Journal of Public Health
Research and Development. 2018; 9(3): 393-398.
50. Hamza LF, Sahi NM, Hameed IH. Analysis of
Methanolic extract of Secondary Metabolites
Released by Candida glabratus Using GC-MS and
Evaluation of its Antimicrobial Activity. Indian
Journal of Public Health Research and Development.
2018; 9(3): 345-351.