Effect of Amoxicillin Antibiotic on Germination Rate and Active Components of Triticumaestivum L. (Wheat) Seeds

Abstract

According to the best germination percentage and its rate of TriticumAestivum L (Wheat) seeds treated with Amoxicillin 50mg/ml gave the highest germination percentage of 100% and the highest germination rate 0.6, also it gave highest scavenging free radicals of DPPH with a mean percentage of 82.07%. The GC-MAS tests for the control model and wheat plant treated with the antibiotic amoxicillin at concentrations of 25 and 50 mg. It was found that only two materials present in the three models under study: such as Phytol and gamma.-Sitosterol.

Full text

ISSN 2664-8075 (Print) & ISSN 2706-5774 (Online)
South Asian Research Journal of Applied Medical Sciences
Abbreviated Key Title: South Asian Res J App Med Sci
| Volume-6 | Issue-4 | Jul-Aug- 2024 | DOI: https://doi.org/10.36346/sarjams.2024.v06i04.004
Copyright © 2024 The Author(s): This is an open-access article distributed under the terms of the Creative Commons Attribution
4.0 International License (CC BY-NC 4.0) which permits unrestricted use, distribution, and reproduction in any medium for non-
commercial use provided the original author and source are credited.
Citation: Sundus Hameed Ahmed, Alyaa Mouhsen Yousif, Hashim Kadhim Mohammed, Rasha Sattam Hameed, Khaled
F. M. Salem (2024). Effect of Amoxicillin Antibiotic on Germination Rate and Active Components of Triticumaestivum
L. (Wheat) Seeds. South Asian Res J App Med Sci, 6(4), 98-105.
98
Original Research Article
Effect of Amoxicillin Antibiotic on Germination Rate and Active Components
of Triticumaestivum L. (Wheat) Seeds
Sundus Hameed Ahmed1*, Alyaa Mouhsen Yousif1, Hashim Kadhim Mohammed1, Rasha Sattam Hameed1, Khaled F. M. Salem2
1Biology Department, College of Science, Mustansiriyah University, Baghdad, Iraq
2Sadat University, Iraq
*Corresponding Author: Sundus Hameed Ahmed Email: drsundusahmed@uomustansiriyah.edu.iq
Biology Department, College of Science, Mustansiriyah University, Baghdad, Iraq
Article History
Received: 07.07.2024
Accepted: 16.08.2024
Published: 20.08.2024
Abstract: According to the best germination percentage and its rate of TriticumAestivum L (Wheat) seeds treated with
Amoxicillin 50mg/ml gave the highest germination percentage of 100% and the highest germination rate 0.6, also it gave
highest scavenging free radicals of DPPH with a mean percentage of 82.07%. The GC-MAS tests for the control model
and wheat plant treated with the antibiotic amoxicillin at concentrations of 25 and 50 mg. It was found that only two
materials present in the three models under study: such as Phytol and gamma.-Sitosterol.
Keywords: Amoxicillin, DPPH, GC-MAS, antibiotic.
INTRODUCTION
TriticumAestivum Lis the most widely grown cereal grain in the world and is essential to agriculture [1-4]. It is
cultivated in 70% of the world's cultivated regions and provides the primary nutrition for 36% of the world's population [5,
6]. Approximately 55% of the carbs and 21% of the calories consumed globally come from wheat [6, 7]. Being cultivated
in a wide range of climatic conditions and surpassing all other grain crops (such as rice, maize, etc.) in terms of productivity
and acreage, it is the most significant grain crop on the planet [8, 9]. Among the cereals, wheat is the most significant
primarily due to its grains, which contain unique protein properties both chemically and physically. It also has additional
beneficial elements, likes (Cu, Mg, Zn, Fe, and P), It is a good source of protein, carbs, and vitamins (riboflavin, thiamine,
niacin, and alpha-tocopherol) [10, 11]. Nevertheless, it has been discovered that essential amino acids like lysine and
threonine are absent from wheat proteins [12-14].
The introduction of new and improved varieties that can give higher yields and perform better across a range of
agro-climatic stresses and circumstances may be a means of improving wheat production and quality [15]. Everyone agrees
that one of the most important aspects of plant breeding is the diversity of germplasm in the breeding material [16, 17].
Antibiotics work by either directly eliminating bacteria or preventing their growth in order to treat illnesses in
both humans and animals [18]. Animals' stomachs do not absorb most antibiotics well, and up to 90% of them may be
eliminated [19]. Some antibiotics are very stable in manure and soil, with residues still detectable a year after application
[20]. These antibiotics may be discharged into the environment by grazing animals or dung. Certain antibiotics have the
potential to last for several years [21]. For instance, tetracycline and sulfadiazine were found at average soil concentrations
of 10–15 µg kg−1 and 32–198 µg kg−1, respectively, in agricultural landscapes with typical land use and manure
fertilization [22]. Antibiotics may then be carried from the farmlands to ditches, streams, and rivers by runoff [23], and to
groundwater by leaching (24) or may be consumed by organisms directly [25]. The potential negative impacts of antibiotics
ingested by agricultural plants on human health have been thoroughly studied [26] but considerably less research has been
done on the impact of antibiotics on plants, especially non-crop species. Antibiotics have been shown to significantly
impair plant growth and performance [27]; on the other hand, they may potentially stimulate allomet-ric responses.

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© South Asian Research Publication, Bangladesh Journal Homepage: http://sarpublication.com 99
The study aims to investigate the following: the impact of the antibiotic amoxicillin on wheat seed germination;
the active ingredients in wheat models treated with the antibiotic and their comparison to the control; and the antibacterial
and antioxidant efficacy of germinated seed extracts.
MATERIALS AND METHODS
The purpose of the study was to ascertain how the antibiotic amoxicillin affected the germination of seeds. Seed
germination prior to germination, the seeds were steeped in 70% ethanol for around two minutes. They were then rinsed
numerous times in sterile water to remove any last traces of alcohol (ethanol), and finally submerged in sterile water
together with amoxicillin (25–50 mg/ml).
The germination rate was calculated according to:
Where GN denotes the number of wheat seeds that germinated and SN is the number of tested wheat seeds. The
equation that follows: Ten seeds each were allowed to germinate for five days at 25 °C in a dark atmosphere on a Petri
plate. The length of the shoot and roots was then measured in order to assess the germination rate.
Extract Preparing
Following a 4-day soak in 400 mL of water (v/v) at room temperature (28 C), all of the germinated seeds from
the treated plant and the control were dried in an oven at 40 C and ground into powder using a grinder. Cheesecloth and
Whatman filter paper were used to filter out the dry leaves. A rotary evaporator was then used to further concentrate the
filtrate. Glass Petri dishes were used to hold all of the extracts. After being dried, the plant extracts were redissolved in
water to create a solution that had 3 mg/mL of extract, which was subsequently utilized for tests.
Activity of Antioxidants (DPPH test)
All extracts were evaluated for their antioxidant activity (free radical scavenging activity) using the 1,1-diphenyl-
2-picrylhydrazyl (DPPH) test. First, 5 mL of a 0.004% (w/v) DPPH in methanol solution was mixed with 50 µL of 3.0
mg/mL extracts (70% methanol as blank). Using the dark box, this was incubated in the dark for 30 minutes. Every sample
that was computed using the following formula underwent the same process:
RESULT AND DISCUSSION
Table 1A presents the percentage of germination, was calculated according to Equation 1 as well as the actual
measurements Based on Table 1A, it is noticed that the best solutions for wheat seed germination are Amoxicillin 50mg/ml.
According to the best germination percentage and its rate, both of them had the highest germination percentage of 100%
and the highest germination rate of 0.6.
Table 1A: Germination percentage and germination rate
No.
Treatment (GP %) GR
Treatment (GP %)
GR
1
Control
84.32
0.4
2
Amoxcilline 50mg/ml
100
0.6
3
Amoxcilline 25mg/ml
58.54
0.34
Figure 1, indicates a significant difference of mean percentage scavenging between all the tested extracts (Treated
wheat plant with 50mg/ml, treated wheat plant with 25mg/ml, 3: Control, 4: Ascorbic Acid. The results showed that extract
of treated wheat plant with 500mg/ml exhibited the greatest free radical scavenging activity among the plant extracts with
a mean percentage of 82.07%. On the other hand, the scavenging activity of treated wheat plant with 250mg/ml and not
treated wheat plant extracts (control) was 52.31%, control 72,18% and Ascorbic Acid 92.86% (standard), respectively. It
should be emphasized that toxic effect of the antioxidant and antimicrobial agent on the host cells must be considered, as
a substance may exhibit antioxidant and antimicrobial activity by virtue of its toxic effect on the cells [22].

Sundus Hameed Ahmed et al, South Asian Res J App Med Sci; Vol-6, Iss-4 (Jul-Aug, 2024): 98-105
© South Asian Research Publication, Bangladesh Journal Homepage: http://sarpublication.com 100
Figure 1: Scavenging effect (%) of plant extracts and known, 1: treated wheat plant with 50mg/ml, 2: treated
wheat plant with 25mg/ml, 3: Control, 4: Ascorbic Acid
To evaluate the scavenging effect of the extracts in this study, DPPH reduction was investigated against positive
controls (Ascorbic acid). The more antioxidants occurred in the extract, the more DPPH reduction will occur. High
reduction of DPPH is related to the high scavenging activity performed by particular sample treated with 50mg/ml
amoxicillinin compare with other treatment [20]. Tables (1 and 2) showed the results of the GC-MAS tests for the control
model of the wheat plant treated with the antibiotic amoxicillin at concentrations of 25 and 50 mg. It was found that new
medicinal substances appeared in treated plant differed from the active ingredients originally present in the control wheat
plant, the highest peaks appeared in GC-MAS tests as shown: Phytol, 35.40% (anticancer, anti-inflammatory,
antimicrobial, diuretic), Methyl 9,12,15-octadecatrienoate, 15.36% (Anti-microbial), Methyl palmitate 9.32% (antioxidant,
hypocholesterolemic, antiandrogenic, Hemolytic [21].
Seed extract treated with amoxicillin 50mg/ml, Phytol, 30.185% (anticancer, anti-inflammatory, antimicrobial,
diuretic), Methyl linolenate, 14.76% Antiinflammatory, Hypocholesterolemic, Cancer preventive Hepatoprotective,
Nematicide, Insectifuge Antihistaminic, AntieczemicAntiacne [22], 5-Alpha reductase inhibitor Antiandrogenic,
AntiarthriticAnticoronary, (9-Octadecenamide, 9.10% (Moisturiser for skin, nails, and hair Hepatoprotective,
antihistaminic, hypocholesterolemic, anti-eczemic [23] (Amoxicilene) seed extract treated with amoxicilene 25mg/ml.
Phytol 32.87%, 9-Octadecenamide, (Z)-, 17.77% (Anti inflammatory, Antiandrogenic, Cancer preventive, Dermatitigenic,
Hypocholesterolemic [24], 5- Alpha reductase inhibitor, Anemiagenic, Insectifuge).
Methyl9,12,15-octadecatrienoate), 11.34% (Antiinflammatory, Hypocholesterolemic, Cancer preventive,
Hepatoprotective, Nematicide, Insectifuge Antihistaminic, Antiarthritic, Anticoronary, AntieczemicAntiacne [25], 5-
Alpha reductase inhibitor Antiandrogenic, It was found that there are two materials present in the three models under study:
such as Phytol and gamma.-Sitosterol [26].
Table 1: GC- Mass of control germinated seed extract
No
RT
(min)
Area
%
Name
Quality
Bioactivity
1
14.189
4.51
2-Propenoic acid, 2-methyl-,
1,2-ethanediyl ester
40
Antimicrobial and antioxidant properties Mujeeb
et al., (2014)
2
26.247
9.32
Methyl palmitate
98
Antioxidant, Hypocholesterolemic,
Antiandrogenic, HemolyticGc15
3
28.8
4.22
9,12-Octadecadienoic acid
(Z, Z)-, methyl ester
99
Moisturiser for skin, nails, and hair3
4
28.857
15.36
Methyl 9,12,15-
octadecatrienoate
98
Antibacterial, anticancer, anticandidal, anti-
inflammatory, hypocholesterolemic,
hepatoprotective,3
5
28.94
0.80
Methyl trans-8-
octadecenoate
96
Anti-microbial 5
6
29.023
0.38
Dihydro-.beta.-agarofuran
43

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© South Asian Research Publication, Bangladesh Journal Homepage: http://sarpublication.com 101
No
RT
(min)
Area
%
Name
Quality
Bioactivity
7
29.215
35.40
Phytol
93
Anticancer, anti-inflammatory, antimicrobial,
diuretic effects Bharathy et al., (2012) 2
Antimicrobial, Anti-inflammatory, Anticancer,
Diuretic, Antifungal against S. typhi, resistant
gonorrhea, joint dislocation, headache, hernia,
stimulant and antimalarial 4
8
29.387
3.21
Octadecanoic acid, methyl
ester
97
antimicrobial activity Kannan & Kannan
(2019) 2Hepatoprotective, antihistaminicGc19
9
34.331
9.79
alpha.-Monopalmitin
43
10
34.419
4.76
1-Heneicosyl formate
81
11
37.444
5.60
Diisooctyl phthalate
16
Antimicrobial, Antifouling4Anti-carcinogenic,
Used in skin treatments and Cosmetic products 5
12
44.558
6.66
gamma.-Sitosterol
91
Anticancer, antimicrobial, antidiabetic,
antifertility, antioxidant 5
GC-Mass for Control germinated seed extract
Table 2: GC- Mass of control germinated seed extract T 50 mg/ml
No
RT
(min)
Area
%
Name
Quality
1
14.173
2.59
2-Methoxy-4-vinylphenol
96
Antimicrobial, antioxidant, anti-inflammatory,
analgesic, anti-germination
2
14.796
2.54
4-Methylproline methyl ester
64
3
16.373
4.63
D-Fructose, 1-O-methyl-
22
4
26.247
7.78
Methyl palmitate
98
Antioxidant, Hypocholesterolemic,
Antiandrogenic, Hemolytic
5
28.8
4.98
9,12-Octadecadienoic acid (Z,
Z)-, methyl ester
99
Moisturiser for skin, nails, and hair
Hepatoprotective, antihistaminic,
hypocholesterolemic, anti-eczemic
6
28.857
14.76
Methyl linolenate
99
Cosmetic, Coloring agent. Antiinflammatory,
Hypocholesterolemic, Cancer preventive
HepatoprotectiveNematicide, Insectifuge
Antihistaminic, AntieczemicAntiacne, 5-Alpha
reductase inhibitor Antiandrogenic,
Antiarthritic, Anticoronary, Insectifuge
7
28.935
0.73
Methyl oleate
87
Antiinflammatory, Antiandrogenic Cancer
preventive, Dermatitigenic

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No
RT
(min)
Area
%
Name
Quality
Hypocholesterolemic, 5-Alpha reductase
inhibitor, Anemiagenic Insectifuge, Flavor
8
29.034
0.25
trans-.delta.(sup 9)-Octadecenoic
acid
91
Emulsifying agent Anti-cancer
9
29.091
0.45
2,5-Diaminobenzophenone
42
10
29.215
30.18
Phytol
90
anticancer, anti-inflammatory, antimicrobial,
diuretic effects
11
29.381
2.24
Octadecanoic acid, methyl ester
98
Anti-microbial 5 Anti-cancer5 Cosmetic,
Flavour, Hypocholesterolemic, Lubricant,
Perfumery, Propecic, Suppository. Gc25
12
32.292
9.10
9-Octadecenamide, (Z)-
87
Anti inflammatory, Antiandrogenic, Cancer
preventive, Dermatitigenic,
Hypocholesterolemic, 5-Alpha
reductaseinhibitor, Anemiagenic, Insectifuge 9
13
36.91
4.14
1,3-dimethyl-4-azaphenanthrene
38
14
38.145
4.31
Bis(2-ethylhexyl) sebacate
50
15
41.606
2.47
Cyclotrisiloxane, hexamethyl-
74
No activity reported 11
16
43.11
2.33
Cyclotrisiloxane, hexamethyl-
72
17
44.553
6.53
gamma.-Sitosterol
91
Anticancer, antimicrobial, antidiabetic,
antifertility, antioxidant 5
GC- Mass of control germinated seed extract T 50 mg/ml
Table 3: GC- Mass of control germinated seed extract T25
No
RT
(min)
Area
%
Name
Quality
1
14.189
5.61
2-Propenoic acid, 2-methyl-,
1,2-ethanediyl ester
47
Antibacterial, flavor 7
2
25.054
1.02
.beta.-Citronellol
55
3
25.48
0.91
Cyclotetradecane
98
4
26.237
5.28
Pentadecanoic acid, 14-
methyl-, methyl ester
98
5
28.8
4.04
Methyl linoleate
99
Antiinflammatory, Hypocholesterolemic,
Cancerpreventive Hepatoprotective
Nematicide, InsectifugeAntihistaminic,
AntieczemicAntiacne, 5-Alphareductase
inhibitorAntiandrogenic, Antiarthritic,

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No
RT
(min)
Area
%
Name
Quality
Anticoronary, InsectifugeGc13
6
28.858
11.34
Methyl 9,12,15-
octadecatrienoate
99
Antiinflammatory, Hypocholesterolemic,
Cancer preventive, Hepatoprotective,
Nematicide, Insectifuge Antihistaminic,
Antiarthritic, Anticoronary,
AntieczemicAntiacne, 5-Alpha reductase
inhibitor
Antiandrogenic 17
7
28.941
0.52
trans-.delta.(sup 9)-
Octadecenoic acid
90
Emulsifying agent, 1Anti-cancer5
8
29.216
32.87
Phytol
90
anticancer, anti-inflammatory, antimicrobial,
diuretic effects Bharathy et al., (2012) 2
Antimicrobial, Anti-inflammatory, Anticancer,
Diuretic, Antifungal against S. typhi, resistant
gonorrhea, joint dislocation, headache, hernia,
stimulant and antimalarial 4can be used as a
precursor for the manufacture of synthetic
forms of vitamin E and vitamin K1gc 12
9
29.76
2.33
Hexadecanamide
94
Antiinflammatory, Antialopecic,
Antifibrinolytic, Lubricant, Antiandrogenic,
flavour, Hemolytic, Antioxidant,
Hypocholesterolemic, Nematicide, Pesticide,5-
Alpa-reductase inhibitor. Ge 25
10
32.292
17.77
9-Octadecenamide, (Z)-
96
Anti inflammatory, Antiandrogenic, Cancer
preventive, Dermatitigenic,
hypocholesterolemic, 5-Alpha
reductaseinhibitor, Anemiagenic, Insectifuge 9
11
34.425
4.56
Cyclohexane, 1-(1,5-
dimethylhexyl)-4-(4-
methylpentyl)-
45
12
37.44
1.66
N-ethyl-1,3-dithioisoindoline
53
13
41.606
5.42
Cyclotrisiloxane, hexamethyl-
64
No activity reported 11
14
44.553
6.66
.gamma.-Sitosterol
91
Anticancer, antimicrobial, antidiabetic,
antifertility, antioxidant 5
GC- Mass of control germinated seed extract T25

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© South Asian Research Publication, Bangladesh Journal Homepage: http://sarpublication.com 104
Table 4 showed comparism between control and treated seed with 50mg/mland25mg/ml, we found that only two
materials present in the three models under study: such as Phytol and gamma.-Sitosterol
Table 4: Comparism between active component of treated Wheat plant samples
No
Control
T.Amox.50/ml
T.Amox.25/ml
1
2-Propenoic acid, 2-methyl-, 1,2-
ethanediyl ester
2-Methoxy-4-vinylphenol
2-Propenoic acid, 2-methyl-, 1,2-
ethanediyl ester
2
Methyl palmitate
4-Methylproline methyl ester
.beta.-Citronellol
3
9,12-Octadecadienoic acid (Z,Z)-
, methyl ester
D-Fructose, 1-O-methyl-
Cyclotetradecane
4
Methyl 9,12,15-
octadecatrienoate
Methyl palmitate
Pentadecanoic acid, 14-methyl-,
methyl ester
5
Methyl trans-8-octadecenoate
9,12-Octadecadienoic acid (Z,Z)-,
methyl ester
Methyl linoleate
6
Dihydro-.beta.-agarofuran
Methyl linolenate
Methyl 9,12,15-octadecatrienoate
7
Phytol
Methyl oleate
trans-.delta.(sup 9)-Octadecenoic
acid
8
Octadecanoic acid, methyl ester
trans-.delta.(sup 9)-Octadecenoic
acid
Phytol
9
.alpha.-Monopalmitin
2,5-Diaminobenzophenone
Hexadecanamide
10
1-Heneicosyl formate
Phytol
9-Octadecenamide, (Z)-
11
Diisooctyl phthalate
Octadecanoic acid, methyl ester
Cyclohexane, 1-(1,5-dimethylhexyl)-
4-(4-methylpentyl)-
12
.gamma.-Sitosterol
9-Octadecenamide, (Z)-
N-ethyl-1,3-dithioisoindoline
13
Control
1,3-dimethyl-4-azaphenanthrene
Cyclotrisiloxane, hexamethyl-
14
2-Propenoic acid, 2-methyl-, 1,2-
ethanediyl ester
Bis(2-ethylhexyl) sebacate
.gamma.-Sitosterol
15
Cyclotrisiloxane, hexamethyl-
Amox.250
16
.gamma.-Sitosterol
2-Propenoic acid, 2-methyl-, 1,2-
ethanediyl ester
17
.beta.-Citronellol
CONCLUSIONS
It was discovered that only two materials were consistently present across all three models under study: Phytol
and gamma-Sitosterol. This finding highlights the commonality of these two compounds despite the variations in the other
components analyzed in each model.
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