SJA_41_3_1455-1462

Introduction:

The genus Allium L. belongs to the family Amaryllidaceae, the subfamily Alliodeae, and the tribe Allieae (Choi et al., 2011). The family consists of 13 genera and 600 species, as indicated (Klopper et al., 2006) The garlic family contains many secondary metabolites, as between Okaka and Okaka (2001), the high nutritional value of garlic, as it contains some antioxidant factors such as phenols, alkaloids, tannins. (Divya et al., 2017) demonstrated that garlic contains 100 secondary metabolites, which are effective due to their chemical properties, including terpenes, phenols and alkaloids. Garlic is a perennial herbaceous plant with bulbs used for medicinal purposes. It contains active components such as allicin and alliin, which are used for a variety of purposes. It is a commonly used spice worldwide and is characterized by its strong, pungent odor, (Okoro et al., 2023). Onions A. cepa contain numerous chemical compounds, such as allicin, quercetin, fisetin, and other sulfurous compounds, such as diallyl disulfide and diallyl trisulfide, (Dorrigiv et al., 2021). It contains many vitamins and secondary organic compounds, the main component of which is sulfur, such as Allyl propyl disulphide (Al-Jasim, 2022). It has a great health benefit because it contains many secondary metabolites such as phenols that help prevent cardiovascular diseases and cancers (Ly et al., 2005). Dorrigiv et al. (2021) mentioned that the benefits of the A. cepa genus also include reducing the toxicity of chemical agents in many organs of the body, such as the liver, brain, kidneys, blood, and others, by reducing lipid peroxidation, removing free radicals, and having an antioxidant effect, it is also an anti-inflammatory.

Ortega et al. (2017) also confirmed that onions are rich in terpenes, as well as that A. ampeloprasum is rich in terpenes, phenols and tannins. As for (Wissam and Mohammed, 2020) he mentioned that Onions, which belong to the Alliaceae family are one of the important crops for the large deities importance of nutrient content as fresh onions contain 1.2 mg of proteins

Abdulla et al. (2013) also confirmed this in his study of leek leaf extract, as it had the highest percentage of phenols. Which to differentiate the Amaryllidaceae species under study can consider all of the above from the clear chemical evidence.

Materials and Methods

Chemical study

Preparation of chemical extracts

The chemical compounds were extracted from the leaf powder under study according to the method adopted by (Markham, 1982) with some modifications as follow:

1. The samples (leaves) were collected and dried at room temperature after being washed and removing any impurities. They were then ground in an electric grinder to obtain a fine powder and stored in a plastic container.

2. Take 1 gram of powder and mix with continuous stirring in a glass tube with 10 ml of 99% methanol solution and leave in a dark place for 12 hours.

3. The extract was then filtered into another glass tube using a filter connected to a medical syringe with a 0.45 μm orifice.

4. To increase the concentration of the extract and remove water, (1) ml of hexane at a concentration of (99%) was added. Due to the presence of hexane, the floating part separated from the water was withdrawn and the chemical compounds were evaluated.

5. Gas chromatography/mass spectrometry (GC/MS) technique was used to identify and separate the chemical compounds from the leaf extracts of the plants under study.

GC-MS analysis method

Using gas chromatography-mass spectrometry (GCMS) on a Hewlett-Barchard gas chromatograph (GC) the essential oil was analyzed. The oven temperature was set from 70 to 240°C at a rate of 5°C/min, the ion source was set at 240°C, and the ionization rate was 70 electrons. The carrier gas used was helium at a rate of 1 ml/min, and the scanning range was between 35 and 425 atomic mass units ,Diluted oil in n-hexane (1.0 μL) was injected into the.(Mohammed et al., 2020)

Chemical compounds and how they were identified

Based on the interpretation of the mass spectrum (GC/MS ), the components were identified. Based on the data of the National Institute of Science and Technology, the spectrum of the resulting unknown components was compared with a set of known components preserved in the library of the above-mentioned institute to determine the name of the test substance, its molecular weight and structure. The test was conducted in the (GC/MS) unit of the Ministry of Science and Technology.

Results and Discussion

The current study observed an abundance of chemical content in the studied species, with 75 chemical compounds recorded, including phenols, terpenes, alkaloids, and esters. The A. cepa leaf extract contained the largest proportion of compounds, amounting to 28 compounds Table 2, while A. sativum contained 27 compounds Table 1, and A. ampeloprasum contained 20 compounds Table 3.

Table 1: GC-MAS analysis of the plant extract of A. sativum L. leaves

Composite type	The sequence	The name of the boat	Chemical formula	Summit number	Running time per minute	Space٪
Phenolic compounds	1	Hexane	C6H14	1	2.037	4.28
	2	Propylene glycol	C3H8O2	2	2.382	1.17
	3	Glycerin	C3H8O3	3	6.206	0.55
	4	(Z)6-Pentadecen-1-ol	C15H30O	13	21.084	9.59
	5	Octadecanoic acid	C18H36O2	14	21.287	7.97
	6	Hexane, 1,6-dibromo	C6H12Br2	16	22.626	1.36
	7	Nonane, 5-methyl-5-propyl	C13H28	22	25.669	0.87
	8	16-Hentriacontanone	C31H62O	27	29.255	1.44
	9	Nonane, 1-iodo	C9H19I	19	24.095	0.56
Turbine compounds	10	n-Hexadecanoic acid	C16H32O2	5	15.296	1.82
	11	Pentadecanoic acid	C15H30O2	9	19.376	40.78
	12	Isobutyl nitrite	C5H11NO2	10	19.921	0.76
	13	Isoamyl nitrite	C4H9NO2	11	20.006	0.21
	1٤	Acetic acid, 2-propenyl ester	C5H8O2	17	23.044	0.14
	15	3-Hexanone, 2,2-dimethyl	C8H16O	18	23.271	0.53
	16	3,7,11-Tridecatrienenitrile, 4,8,12-trimethyl	C16H25N	23	26.744	1.03
	17	3,5-Dimethyl-4-octanone	C10H20O	25	27.930	16.60
Alkaloid compounds	18	3-Isopropylbenzaldehyde	C10H12O	4	10.231	1.00
	19	Undecanol-4	C11H24O	12	20.283	1.29
	20	1-Iodoundecane	C11H23I	15	22.395	0.38
	21	Decane, 6-ethyl-2-methyl	C13H28	21	24.897	0.44
	22	n-Decanoic acid	C10H20O2	7	18.216	0.95
	23	2-Propanone, 1,1,1-trifluoro	C3H3F3O	8	19.101	0.24
Esters	24	Tridecane, 1-iodo	C13H27I	26	28.191	0.22
	25	Acetic acid, 2-propenyl ester	C5H8O2	6	17.808	0.07
	26	1,2-Benzenedicarboxylic acid, diisooctyl ester	C24H38O4	20	24.577	2.07
	27	Sulfurous acid, butyl 2-ethylhexyl ester	C12H26O3S	24	27.25	0.77
100.00

Table 2: GC-MAS analysis of plant extract of A. cepa leaves

Composite type	The sequence	The name of the boat	Chemical formula	Summit number	Running time per minute	Space ٪
Phenolic compounds	1	Hexane	C6H14	1	2.037	4.70
	2	Pentanal, 2-methyl	C6H12O	3	13.766	0.77
	3	(Z)6,(Z)9-Pentadecadien-1-ol	C15H28O	13	21.156	22.20
	4	Decane, 6-ethyl-2-methyl	C13H28	19	24.899	0.57
	5	Nonane, 1-iodo	C9H19I	21	26.430	0.39
	6	2,7-Nonadien-5-one, 4,6-di methyl-	C11H18O	22	26.745	0.75
	7	Heptadecane, 2,6,10,15-tetramethyl	C21H44	23	27.269	7.26
	8	2-Methyl-3-decanol	C11H24O	24	27.911	0.22
	9	2H-1-Benzopyran-6-ol, 3,4-dihydro-2,8-dimethyl-2-(4,8,12-trimethyltridecyl	C27H46O2	25	28.019	9.62
Turbine compounds	10	Ethene, methoxy	C3H6O	2	10.928	0.23
	11	1H-Imidazolecarboxylic acid-, (1-methylethyl) ester	C7H10N2O2	5	17.884	0.25
	1٢	Butanoic acid, anhydride	C8H14O3	8	19.064	0.41
	1٣	n-Hexadecanoic acid	C16H32O2	9	19.356	23.60
	1٤	Hexanoic acid, 2,2-dimethyl	C8H16O2	10	19.488	0.15
	15	phytol	C20H40O	12	20.745	1.30
	16	Hexadecane	C16H34	17	24.10	2.59
	١7	Pentadecana	C15H30O	27	28.687	4.57
	١8	gamma.-tocopherol	C28H48O2	28	29.131	1.57
Alkaloid	١9	Tetratetracontane	C44H90	20	25.678	8.06
	20	3,5-Dimethyl-4-octanone	C10H20O	26	28.190	0.20
	٢1	4-(2-Ethylpiperidin-1-yl)butironitrile	C9H19I	29	29.272	7.38
Esters	٢2	Isobutyl nitrite	C4H9NO2	4	17.104	0.15
	٢3	Phthalic acid, 4-bromophenyl heptyl ester	C21H23BrO4	7	18.256	0.25
	٢4	Oxalic acid, butyl propyl ester	C9H16O4	11	20.541	0.07
	٢5	Heptadecanoic acid, heptadecyl ester	C34H68O2	14	21.274	1.02
	٢6	Oxalic acid, isobutyl pentyl ester	C11H20O4	15	22.395	0.21
	٢7	Oxalic acid, 2-ethylhexyl hexyl ester	C16H30O4	16	23.275	0.64
	٢8	Phthalic acid, heptyl 2-pentyl ester	C20H30O4	18	24.579	0.55

The distinctiveness of each species with chemical compounds gave it great taxonomic importance. Record 27 phenolics. All species contained 9 phenolic compounds, the highest percentage of phenols was recorded in A. sativum, for (Z) 6-Pentadecen-1-ol, it reached 9.59% per minute 21.084. The highest rate was recorded at 22.20% for (Z) 6, (Z) 9-Pentadecadien-1-ol in the species A. cepa, as it recorded an onset time in the 21.15 minute, while the compound Hexane recorded the highest percentage in the extract of A. ampeloprasum leaves. If it reached 14.87% in the minute 2,098. The study agreed with (Fomina and Kukushkina, 2020) that the Allium genus is one of the plants that contain phenolic compounds.

While 22 terpens compounds were found in the species under study, the largest share was in A. cepa, which recorded 9 terpens compounds . The n-Hexadecanoic acid recorded the highest percentage of 23.60% per minute 19.356. While the extract of A. sativum leaves contained 8 compounds, the highest percentage was 40.78% per minute 19.376 for the compound Pentadecanoic acid, while the A. ampeloprasum species contained only 5 compounds that recorded the highest percentage of compound Z, Z-8,10-Hexadecadien-1-ol if reached 34.56% per minute 21.096.

Also, 11 of the alkaloid compounds were identified in the studied species, the largest share of them was in the species A. sativum if it reached 6 compounds, and the highest percentage was 1.29% per minute for Undecanol-4, while the species Allium cepa contained 3 compounds. The highest percentage of which was for the compound Tetratetracontane when it reached 8.06% per minute 25.678, and only two compounds of alkaloids in the species A. ampeloprasum A. ampeloprasum A. ampeloprasum A. ampeloprasum A. ampeloprasum had the highest percentage of the compound Decane, 6-ethyl-2-methyl, if it was 0.54% per minute 24.075.

Table 3: GC-MAS analysis of the plant extract of A. ampeloprasum leaves

Composite type	Composite type	The name of the boat	Chemical formula	Summit number	Running time per minute	Space ٪
Phenolic compounds	1	Hexane	C6H14	1	2.098	14.87
	2	Phenol, 2-methoxy-4-(1-propenyl)-, acetate	C12H14O3	2	11.966	0.43
	3	1,4-S,S-2,5-Bis[carbethoxy]phenylene bis[N,N-dimethyldithiocarbamate]	C18H24N2O4S4	6	19.462	0.07
	4	2-Heptanone, 6-methyl-5-methylene	C9H16O	10	23.523	0.23
	5	Dodecane, 2,6,10-trimethyl	C5H12	14	25.514	0.31
	6	4-Hexen-3-one	C9H16O4	17	26.219	0.15
	7	Heptadecane, 2,6,10,15-tetramethyl	C9H19NO3	19	27.239	1.84
	8	16-Heptadecenal	C21H44	20	27.875	0.64
	9	16-Hentriacontanone	C15H30O	21	29.244	6.03
Turbine compounds	10	Pentanal, 2-methyl	C6H12O	4	17.927	0.48
	11	n-Hexadecanoic acid	C16H32O2	5	19.285	11.19
	12	Z,Z-8,10-Hexadecadien-1-ol	C16H30O	8	21.096	34.56
	13	cis-9,10-Epoxyoctadecan-1-ol	C18H36O2	9	21.244	0.50
	14	1-Hexene, 4,4-diethyl	C15H32	15	25.650	1.54
Alkaloid	15	Decane, 6-ethyl-2-methyl	C13H28	12	24.075	0.54
Alkaloid	16	Neopentane	C12H20O4	13	25.369	0.23
Esters	17	1H-Imidazolecarboxylic acid-, (1-methylethyl) ester	C7H10N2O2	3	17.857	0.10
	18	Oxalic acid, cyclobutyl heptyl ester	C13H22O4	11	23.941	0.57
	19	Oxalic acid, butyl propyl ester	C10H20	16	25.746	0.46
	20	Nitric acid, nonyl ester	C6H10O	18	26.722	0.64
100.00

Table 4: The chemical compounds common to the species under study

The sequence	The name of the boat	species
The sequence	The name of the boat	A. sativum	A. Cepa	A. ampeloprasum
1	Hexane	+	+	+
2	Decane, 6-ethyl-2-methyl	+	+	+
3	n-Hexadecanoic acid	+	+	+
4	(Z)6-Pentadecen-1-ol	+	+	-
5	Nonane, 1-iodo	+	+	-
6	3,5-Dimethyl-4-octanone	+	+	-
7	Oxalic acid, butyl propyl ester	-	+	+
8	Heptadecane, 2,6,10,15-tetramethyl	-	+	+
9	Pentanal, 2-methyl	-	+	+
10	16-Hentriacontanone	+	-	+

(-) Absence of the compound (+)Presence of a compound

As for the esters, 15 compounds were identified, four of which were found in A.sativum and A. ampeloprasum. The highest percentage was for 1,2-Benzenedicarboxylic acid, diisooctyl ester in A. sativum leaf extract, reaching 2.07% at 24.577 minutes (Figure 1). The highest percentage was for Nitric acid, nonyl ester in A. ampeloprasum , reaching 0.64 at 26.722 minutes (Figure 2). As for A. cepa, seven compounds were identified, with the highest percentage reaching 1.02% at 21.274 minutes for Heptadecanoic acid, heptadecyl ester (Figure 3).

From the aforementioned results, we find that the current study agreed with the study of Divya et al. (2017) and Mikail, (2010) that the species A. sativum contains most of the basic secondary metabolites represented by phenols, terpenes and alkaloids. The study did not agree with Ameh et al. (2013) who confirmed the absence of terpenes in this species. The current study also did not agree with what Boukeria et al. (2016) stated that A. sativum contains only alkaloids and glycosides, while it agreed with the same study that A. cepa contains phenols, terpenes and alkaloids. The current study was consistent with what was stated by Abd and Ali (2013) that A. ampeloprasum contains most of the secondary metabolites such as phenols, terpenes and alkaloids.

The study showed the presence of 10 compounds common to the species under study, Table 4, as the compounds Hexane and Decane, 6-ethyl-2-methyl and n- Hexadecanoic acid were found in all species, indicating that the species are chemically close to each other and confirming that they belong to the same family. This supports what was mentioned by Al-Saadiy, (2018) that plants that share chemical compounds indicate a kinship between them. Some compounds were also found in one species and not in another or vice versa, such as the distinction and uniqueness of the species A. cepa by the loss of the compound 16-Hentriacontanone, which was found in the other two species. While the compounds Oxalic acid, butyl propyl ester and Heptadecane, 2,6,10,15-tetramethyl and Pentanal, 2-methyl were unique to the species A. sativum. The species A. ampeloprasum is characterized by the presence of the compounds (Z)6-Pentadecen-1-ol Nonane, 1-iodo, and 3,5-Dimethyl-4-octanone. These chemical compounds can be used to distinguish between these two species and also from a third species, which is of great taxonomic importance in separating the species from one another.

Conclusions

From the current study, it can be concluded that the onion genus contains numerous phenolic and terpene chemical compounds, as well as alkaloids and esters. It is richer in phenolic compounds than other chemical compounds. A. sativum was distinguished by the presence of alkaloid compounds to a greater extent than the other two species under study. The study also revealed that all species share a large number of compounds, which indicates and confirms their close relationship. We also conclude that each species is distinguished by compounds that are unique to it, such as A. cepa, which is distinguished by the presence of the compound 16-hentriacontanone, while the other species lack it.

Recommendations

The current study recommends conducting similar studies on other vegetative and floral plant parts, as well as monitoring the variation in plant chemical compound production across different growth stages and cultivation conditions.

Acknowledgements

The authors would like to thank College of Education, Department Biology and Al-Nasr High School, Al-Diwaniyah Education for their support and providing tools for our experiments

Novelty Statement

An abundance of chemical content, with 75 compounds recorded, distributed among phenolics, terpenes, alkaloids, and esters. These compounds varied depending on their retention time and concentrations among the studied species. A. cepa was characterized by the loss of 16-Hentriacontanone, which was present in the other two specie

Author’s Contribution

Azhar Abdulameer Sosa: Article idea and writing.

Hussein Ali Kadhim Al-Aridhee: Collect data from the field and writing first dirft.

Suhaila Hussein Baji: Article idea and data analysis

Generative AI and AI-assisted technology statement

The authors stated that they didn’t use generative AI and AI-assisted technology in preparing this manuscript.

Conflict of interest

The authors have no conflict of interest.

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