matricaria chamonbmilla: bioactive compounds of methanolic
fruit extract using gc-ms and ftir techniques and
determination of its antimicrobial properties
rafid hadi hameed1, ghaidaa jihadi mohammed2, imad hadi hameed3
1ministry of health, mesan health office, mesan 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
matricaria chamomilla is the most popular source of the herbal product chamomile, although other species
are also used as chamomile. the objectives of our study were analysis of the secondary metabolite products.
nineteen bioactive compounds were identified in the methanolic extract of matricaria chamomilla. the
identification of bioactive chemical compounds is based on the peak area, retention time molecular weight
and molecular formula. gc-ms analysis of matricaria chamomilla revealed the existence of the 4-amino-
1,5-pentandioic acid , (r)-lavandulyl acetate , 1,3-diazacyclooctane-2-thione , glucosamine , n-acetyl-nbenzoyl-
, trans – traumatic acid , dodecanoic acid , 3-hydroxy- , cyclopentanone , 2-cyclopentylidene- , (e)-
?-famesene , 1-naphthalenol , 2-methyl- , pregn-4-ene,3,20-dione ,17,21-dihydroxy-,bis(o-methyloxime) ,
9,12,15-octadecatrienoic acid ,2,3-dihydroxypropyl ester , (z) , pyrazole[4,5-b]imidazole,1-formyl-3-ethyl-
6-?-d-ribofuranosyl- , 1,6-dioxaspiro[4. 4]non-3-ene,2-(2,4-hexadiynylidene)- , 9,12-octadecadienoic acid
(z,z)- , 9-octadecenamide,(z)- , 1,2-benzenedicarboxylic acid , bis(8-methylnonyl) ester , carda-4,20(22)-
dienolide , 3-[(6-deoxy-3-o-methyl-?-d-allopyra and campesterol. maximum zone formation was against
proteus mirabilis 6.01±0.23. matricaria chamomilla was very highly active against aspergillus terreus
5.89±0.24.
keywords: matricaria chamomilla, bioactive compounds, antimicrobial, methanol, fruit, gc-ms.
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
matricaria chamomilla has a branched, erect and
smooth stem, which grows to a height of 15–60 cm
(6–23.5 in)1. the long and narrow leaves are bipinnate
or tripinnate. the flowers are borne in paniculate flower
heads (capitula). the white ray florets are furnished with
a ligule, while the disc florets are yellow 2-7. the hollow
receptacle is swollen and lacks scales. it can be taken as
an herbal tea, two teaspoons of dried flower per cup of
tea, which should be steeped for 10 to 15 minutes while
covered to avoid evaporation of the volatile oils. for a sore
stomach, some recommend taking a cup every morning
without food for two to three months8-13. m. chamomilla
can be found near populated areas all over europe and
temperate asia, and it has been widely introduced in
temperate north america and australia8-31. it often grows
near roads, around landfills, and in cultivated fields as
a weed, because the seeds require open soil to survive.
one of the active ingredients of its essential oil is the
terpene bisabolol 14-27. other active ingredients include
farnesene, chamazulene, flavonoids (including apigenin,
quercetin, patuletin and luteolin) and coumarin28-37. dried
chamomile has a reputation (among herbalists) for being
incorrectly prepared because it is dried at a temperature
above the boiling point of the volatile components of
the plant 38-39. the aims of this study were analysis of
the secondary metabolite products and evaluation of its
antibacterial and antifungal activities.
doi number: 10.5958/0976-5506.2018.00213.9
224 indian journal of public health research & development, march 2018, vol.9, no. 3
material and method
gas chromatography–mass spectrum: matricaria
chamomilla gc–ms analysis were carried out in a
gc system (agilent 7890a series, usa). the column
temperature was kept at 40?c for 1 min followed by
linear programming to raise the temperature from 40?to
120?c (at 4 c? min?1with 2 min hold time), 120 c? to
170 c? (at 6 c? min?1with 1 min hold time) and 170 c?
to 200 c? (at10?c min?1with 1 min hold time)40.
fourier transform infrared spectrophotometer
(ftir): the powdered matricaria chamomilla was
treated for fourier transform infrared spectrophotometer
(shimadzu, ir affinity 1, japan). the sample was run at
infrared region between 400 nm and 4000 nm41.
determination of antibacterial and antifungal
activity of crude bioactive compounds of matricaria
chamomilla: the test bacterial pathogens were swabbed
in müller-hinton agar plates. sixteen ml of matricaria
chamomilla extract was loaded on the bored wells. the
wells were bored in 0.5 cm in diameter 42. the plates
were incubated at 37?c for 24 h and examined. after
the incubation the diameter of inhibition zones around
the discs was measured. five-millimeter diameter wells
were cut from the agar using a sterile cork-borer, and
fifty ?l of the samples solutions matricaria chamomilla
was delivered into the wells. antimicrobial activity was
evaluated by measuring the zone of inhibition against
the test microorganisms. methanol was used as solvent
control. 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 incubation43-46.
statistical analysis: 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.
table 1: major phytochemical compounds identified in methanolic extract of matricaria chamomilla
pharmacological
exact mass actions molecular
phytochemical compound rt (min) weight s.
no.
anti-proliferative
1. 4-amino-1,5-pentandioic acid 4.185 175 175.084458 activity
2. (r)-lavandulyl acetate 4.472 196 196.14633 antioxidant activity
3. 1,3-diazacyclooctane-2-thione 5.576 144 144.07212 unknown
4. glucosamine , n-acetyl-n-benzoyl- 5.851 325 325.116152 unknown
5. trans – traumatic acid 6.182 228 228.136159 anti-hyperalgesic
6. dodecanoic acid , 3-hydroxy- 7.338 216 216.1725445 anti-hiv activity
anti-inflammatory
7. cyclopentanone , 2-cyclopentylidene- 7.951 150 150.1044655 activity
8. (e)-?-famesene 8.374 204 204.1878 anti-infective
9. 1-naphthalenol , 2-methyl- 8.443 158 158.073165 anti-inflammatory
anti-inflammatory
10.153 404 404.267508 activity
pregn-4-ene,3,20-dione
,17,21-dihydroxy-,bis(omethyloxime)
10.
antiviral and anti10.972
352 352.26136 obesity properties 9,12,15-octadecatrienoic acid
11. ,2,3-dihydroxypropyl ester , (z)
pyrazole[4,5-b]imidazole,1-formyl-3- 12.002 296 296.11207 anti-pain compound
12. ethyl-6-?-d-ribofuranosylanti-
inflammatory,
12.980 200 200.08373 analgesic, antipyretic 1,6-dioxaspiro[4.4]non-3-ene,2-(2,4-
13. hexadiynylidene)-
14. 9,12-octadecadienoic acid (z,z)- 15.406 280 280.24023 anti-inflammatory
anti-inflammatory
15. 9-octadecenamide,(z)- 17.317 281 281.271864 antibacterial activity
indian journal of public health research & development, march 2018, vol.9, no. 3 225
contd…
1,2-benzenedicarboxylic acid , bis(8- 21.391 446 446.33961 anti-inflammatory
16. methylnonyl) ester
carda-4,20(22)-dienolide , 22.158 548 548.298534 antibacterial activity
17. 3-[(6-deoxy-3-o-methyl-?-d-allopyra
anti-inflammatory
18. campesterol 23.531 400 400.370516 activity
result s and discussion
identification of biochemical compounds: analysis
of compounds was carried out in methanolic extract
of matricaria chamomilla, shown in table 1.
chromatogram gc-ms analysis of the methanol extract
of matricaria chamomilla 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 4-amino-1,5-pentandioic acid
, (r)-lavandulyl acetate , 1,3-diazacyclooctane-2-
thione , glucosamine , n-acetyl-n-benzoyl- , trans
– traumatic acid , dodecanoic acid , 3-hydroxy- ,
cyclopentanone , 2-cyclopentylidene- , (e)-?-
famesene , 1-naphthalenol , 2-methyl- , pregn-4-
ene,3,20-dione ,17,21-dihydroxy-,bis(o-methyloxime) ,
9,12,15-octadecatrienoic acid ,2,3-dihydroxypropyl ester
, (z) , pyrazole[4,5-b]imidazole,1-formyl-3-ethyl-6-?-
d-ribofuranosyl-, 1,6-dioxaspiro[4.4]non-3-ene,2-(2,4-
hexadiynylidene)-, 9,12-octadecadienoic acid (z,z)- ,
9-octadecenamide,(z)- , 1,2-benzenedicarboxylic acid,
bis(8-methylnonyl) ester , carda-4,20(22)-dienolide,
3-[(6-deoxy-3-o-methyl-?-d-allopyra and campesterol.
the ftir analysis of matricaria chamomilla leaves
proved the presence of alkyl halides, alkenes, amide,
alkanes, and aldyhides which shows major peaks at
667.37, 756.10, 869.90, 889.18, 947.05, 1018.41,
1091.71, 1190.08, 1199.72, 1317.38, 1608.63, 1721.65,
1732.08, 2330.01, 2358.94, 2461.17, 2820.28, 2904.80
and 2929.87. in our research, the anti-microbial activity
of matricaria chamomilla extract was determinate by
determining the zone of inhibition against eight bacteria
and eight fungi and yeast. clinical pathogens were
selected for antibacterial activity namely, streptococcus
pneumonia, pseudomonas eurogenosa, salmonella
typhi, staphylococcus epidermidis, bacillus subtilis,
escherichia coli, proteus mirabilis, and streptococcus
pyogenes. maximum zone formation was against proteus
mirabilis 6.01±0.23. methanolic extraction of matricaria
chamomilla showed notable antifungal activities
against trichoderma horzianum, candida albicans,
aspergillus niger, aspergillus terreus, saccharomyces
cerevisiae, microsporum canis, trichoderma viride,
and penicillium expansum. matricaria chamomilla was
very highly active against aspergillus terreus 5.89±0.24.
in comparison to the antibiotics used in this study, the
plants extracts were far more active against the test
bacterial strains47.
conclusion
medicinal property of matricaria chamomilla
methanolic extract is due to presence of secondary
metabolites. eleven bioactive chemical compounds
were identified by gc-ms analysis. this matricaria
chamomilla 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: all our protocols were approved
under the department of biology, college of science
for women, university of babylon, hillah city, iraq and
all experiments were carried out in accordance with
approved guidelines.
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