Gas Chromatography-Mass Spectrometry Analysis of Chemical Compounds in Ethanolic Extracts of Guiera senegalensis and Geigeria alata

Received 13 Mar, 2024

Accepted 20 Jul, 2024

Published 30 Sep, 2024

Background and Objective: Plants are considered as the primary source of drugs for humans. Plants are rich sources of primary and secondary metabolites. Over the previous decades, plants possessed biologically active metabolites, which have been proven powerful natural medicines. The objective of this research work was to explore the compounds present in the ethanolic extracts of two important Sudanese plant species, namely Guiera senegalensisand Geigeria alataby implicating Gas Chromatography-Mass Spectroscopy (GC/MS) technique. Materials and Methods: The powdered plants samples were extracted with ethanol using a hot extraction technique. The qualitative and quantitative analysis of the samples were carried out by using Gas Chromatography/Mass Spectrometry (GC/MS). Results: The main components of the ethanolic extract of the Guiera senegalensis were 1,2,3-benzenetriol (21%), ethyl alpha-d-glucopyranoside (16.72%), 4H-1-benzopyran-4-one, 2-(3,4-dihydroxy (12%) and quinic acid (11.73%). The main components of the ethanolic extract of Geigeria alatawere azuleno[6,5-b]furan-2,6(3H,4H)-dione,3a (26.69%), n-hexadecanoic acid (8.72%) and 1,3-propanediol, 2-(hydroxymethyl)-2-nitro (5.90%). Conclusion: Guiera senegalensisis a rich source of sugars, alcohols and ketones whereas G. alatais a rich source of ketones, fatty acids, alcohols and steroidal compounds.

INTRODUCTION

Plants are considered as the primary source of drugs for humans. Plants are rich sources of primary and secondary metabolites. Over the previous decades, plants possessed biologically active metabolites, which have been proven powerful natural medicines.

Guiera senegalensis L., a plant of the family Combretaceae, commonly known as “Gubeish”, has played a very important role in traditional medicine in Africa including West Central Sudan, the leaves can be used as medicine to treat gastrointestinal disorders, cough and topically for wound healing. The leaves contain proanthocyanidins: (-)-epicatechin and (-)-epigallocatechin units. The ethanolic extract contains flavonoids, alkaloids, tannins, phenols, saponins, coumarins and terpenoids^1-3. The studies of chemical composition showed that the main chemical components were beta-carboline alkaloids, harman, tetrahydroharman, guieranone A (a naphthyl butanone) it was exhibited potent antifungal activity against Cladosporium cucumerinum. Ethyl acetate fraction of Guiera senegalensis extract found to exhibit fungicidal property⁴.

Methanolic extract of G. senegalensis extract contain the sugars, phospholipids, phytosterols, guieranone A, porphyrin-containing compounds and phenolics⁵. Alkaloidal extract, beta-carboline from G. senegalensis leaves presented an interesting antiplasmodial activity associated with a low cytotoxicity. This findings justified to traditional use of the plant to treat malaria⁶. Investigations on G. senegalensis have demonstrated to possess anti-inflammatory, antioxidant, antiasthmatic, anticancer and antimicrobial^1,2.

Geigeria alata, a plant of the family Asteraceae is commonly known as “Gud-gat”. It has been used in West Central Sudan as a drug to treat diabetes, cough, epilepsy and intestinal complaints, antispasmodic, rheumatism and antihypertension^7-9. Geigeria alata aerial parts essential oil contains alpha-longipinene, beta-caryophyllene, alpha-oxobisabolene and caryophyllene oxide¹⁰. The oil showed moderate in vitro cytotoxicity against some tumor cells¹¹. The Trans-3,5-dicaffeoylquinic acid from Geigeria alata roots is slightly toxic and is a potent alpha-glucosidase inhibitor, reduces blood glucose and blood pressure in diabetic hypertension rats and prevents histopathological changes related to diabetes and hypertension⁹. A sesquiterpene lactone, geigerianolide and flavonoids such as axillarin, quercetin and hispidulin were isolated from G. alata¹². Acyquinic acids in G. alata exhibit antioxidant and antimicrobial capacity¹³.

Aqueous extract of G. alata after 14 days did not produce any significant change (p<0.05) on haematological parameters tested relative to their respective control group¹⁴. Analysis of two medicinal plants, Guiera senegalensis and Geigeria alata growing wild under Sudanese environmental conditions is important as there is little or no information on their chemical composition of ethanolic extracts.

In this study, the GC/MS technique was used, which is useful in detecting the compounds in studied plants and concentration distribution. The present investigation was carried out to determine the possible chemical components from ethanolic extracts of candidate plants using gas chromatography/mass spectrometry.

MATERIALS AND METHODS

Study area: This study was designed and carried out in the Department of Botany, Faculty of Science and Technology, Omdurman Islamic University, Omdurman, Sudan between November, 2016 and March, 2017.

Plant material: Plants samples, Guiera senegalensis and Geigeria alata were purchased from Omdurman Local Market in November, 2016. The voucher specimens were at Herbarium (Ref No. 430 and 512, respectively) of Department of Botany, Faculty of Science and Technology, Omdurman Islamic University.

Preparation of ethanolic extracts: The powdered samples were extracted with ethanol using a hot extraction technique. The powder was extracted separately with 50 mL ethanol by gentle reflux for 1 hr. After extraction, it was filtered and then the removal of solvent was done by a rotary evaporator. Then the (15 mL) was kept in a glass container.

Gas Chromatographic-Mass Spectrometry (GC/MS) analysis: The GC was equipped with MS (Shimatzo QP 2010 GC/MS instrument equipped with reference libraries). Packed material for column were 50% phenyl and 50% methyl polysiloscane, column length 30 m, diameter 0.025 mm, the flow rate of helium as carrying gas was 1 mL/min, the temperature of program consisted of 60-270°C , at rate of 4°C/min. The MS were taken at ionization voltage 70 EV. Library Search was carried out using Wiley GC/MS library. Interpretation on mass spectrum GC/MS was conducted using the database of National Institute Standard and Technology (NIST). The spectrum of the unknown component was compared with the spectrum of the known components stored in the NIST library. The retention time (TR), name of compound and area (%) of the components of the test materials were ascertained.

RESULTS AND DISCUSSION

GC/MS Analysis of Guiera senegalensis: Table 1 showed twenty nine compounds were identified from G. senegalensis. The 1,2,3-Benzenetriol (21%), Ethyl alpha-d-glucopyranoside (16.72%), 4H-1-Benzopyran-4-one, 2-(3,4-dihydroxy (12%), quinic acid (11.73%), sorbitol (7.49%) and n-hexadecanoic acid (5.06%) as the major phytochemical constituents.

GC/MS analysis of Geigeria alata: Table 2 showed twenty nine compounds were identified from G. alata. Azuleno[6,5-b]furan-2,6(3H,4H)-dione,3a (26.69%), n-hexadecanoic acid (8.72%), 1,3-propanediol, 2-(hydroxymethyl)-2-nitr (5.90%), beta-sitosterol (5.05%), acetic acid, 6,8,9-trimethyl-4-pentyl-3-oxa (5.02%) and 8-isopropenyl-1,3,3,7-tetramethyl-bicyclo (2.50%) as the major phytochemical constituents.

The results showed that six compounds in extracts of the two plants contained the same chemical composition, which was 2,4-dihydroxy-2,5-dimethyl- 3(2H)-furan-3-one, 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2-methoxy-4-vinylphenol, 1,2,3-benzenetriol (catechol), n-hexadecanoic acid and squalene, but the percentage content was different.

Table 1:

Chemical constituents of Guiera senegalensis leaves ethanolic extract

Compound name	RT	Area (%)
2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one	4.039	0.67
7-Tridecanone	4.882	0.83
2,5-Dimethyl-4-hydroxy-3(2H)-furanone	5.49	0.38
4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-	6.346	3.18
1,2,3-Propanetriol, 1-acetate	7.809	0.98
2-Methoxy-4-vinylphenol	8.672	0.64
1,2,3-Benzenetriol	10.022	21.02
Quinic acid	12.665	11.73
Ethyl alpha-d-glucopyranoside	12.766	16.72
2-Amino-3-hydroxypyridine	14.08	0.29
7-Oxabicyclo[4.1.0]heptane, 1-methyl-4-(2-methyloxiranyl)-	14.3	0.29
3,7,11,15-Tetramethyl-2-hexadecen-1-ol	14.736	0.77
2-Pentadecanone, 6,10,14-trimethyl-	14.821	0.37
9-Eicosyne	14.994	0.23
Phytol, acetate	15.183	0.35
Sorbitol	15.716	7.49
n-Hexadecanoic acid	16.012	5.06
Hexadecanoic acid, ethyl ester	16.289	1.31
Phytol	17.449	5.05
Oleic acid	17.699	3.28
Octadecanoic acid	17.89	0.41
Ethyl oleate	17.923	0.44
Octadecanoic acid, 2,3-dihydroxypropyl ester	20.846	1.06
1H-Indole-3-carboxylic acid, 5-(acetyloxy)-1-butyl-2-methyl-, ethyl ester	20.969	0.5
Andrographolide	21.236	0.47
2-(1-Methoxynaphthyl-2)quinoline	21.522	0.93
4H-1-Benzopyran-4-one, 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-6-methoxy-	21.652	12
Gamma-sitosterol	21.863	1.79
Squalene	23.158	1.76
RT: Retention time

Table 2:

Chemical constituents of Geigeria alata aerial parts ethanolic extract

Compound name	RT	Area (%)
2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one	4.04	0.37
4,5-Dimethyltetrahydro-1,3-oxazine-2-thione	6.286	0.53
4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-	6.361	0.44
Octanoic acid, ethyl ester	6.911	0.12
Catechol	7.524	0.81
2-Methoxy-4-vinylphenol	8.679	0.56
Decanoic acid, ethyl ester	9.593	0.21
1,6-Cyclodecadiene, 1-methyl-5-methylene-8-(1-methylethyl)-, [S-(E,E)]-	10.112	0.27
1,3-Propanediol, 2-(hydroxymethyl)-2-nitro-	10.527	5.9
1H-Indene, 2,3-dihydro-1,1,5,6-tetramethyl-	10.951	0.3
Phenol, 3,5-bis(1,1-dimethylethyl)-	11.175	0.1
1,2,3,5-Cyclohexanetetrol, (1.alpha.,2.beta.,3.alpha.,5.beta.)-	12.663	1.79
Ethyl alpha-d-glucopyranoside	12.786	1.66
Cyclopentene, 1,2-dimethyl-4-methylene-3-phenyl-	13.86	0.72
Naphthalene, 1,1'-methylenebis[decahydro-	13.926	1.63
n-hexadecanoic acid	16.017	8.72
6-(p-Tolyl)-2-methyl-2-heptenol, trans-	16.29	0.95
Benzene, 1-methyl-4-(2-propenyl)-	16.769	1.16
Linoleic acid ethyl ester	17.675	1.63
Acetic acid, 6,8,9-trimethyl-4-pentyl-3-oxabicyclo[3.3.1]non-6-en-1-ylmethyl ester	17.701	5.02
Epibolin	17.784	1.46
Octadecanoic acid	17.895	1.65
Beta-nootkatol	18.045	2.06
Azuleno[6,5-b]furan-2,6(3H,4H)-dione, 3a,7,7a,8,9,9a-hexahydro-3,5,8-trimethyl-, [3R-(3.alpha.,3a.alpha.,7a.beta.,8.beta.,9a.a	18.838	26.69
8-Isopropenyl-1,3,3,7-tetramethyl-bicyclo[5.1.0]oct-5-en-2-one	19.437	2.5
1,1'-Bis(cyclooct-2-en-4-one)	19.977	2.24
Azuleno[6,5-b]furan-2,6(3H,4H)-dione, 3a,7,7a,8,9,9a-hexahydro-4-hydroxy-3,5,8-trimethyl-, [3R-(3.alpha.,3a.alpha.,4.alpha.,7a	20.843	24.47
Beta-sitosterol	21.872	5.05
Squalene	23.159	0.99
RT: Retention time

Two sugar compounds were identified in the ethanolic extract of G. senegalensis (24.21%). The results revealed that ethyl alpha d-glucopyranoside (16.72%) was formed as a major component. Three alcoholic compounds were identified (22%) and ketonic compounds (13%) were identified.

Five ketonic compounds were identified in the ethanolic extract of G. alata (56.7%). The results revealed that Azuleno-dione (26.69%) was formed as a major component. Six fatty acids were identified (17.35%), three alcoholic compounds (8.64%) and two steroidal compounds were identified 8.64 and 6.04%, respectively.

The biological activities of some components, n-hexadecanoic acid (palmitic acid) possesses some activity like antioxidant, hypercholesterol, nematicidal and pesticide¹⁵. Gamma-sitosterol was previously reported to possess antihyperglycemic activity by increasing insulin secretion in response to glucose¹⁶. Linoleic acid was found to possess antibacterial activity against Bacillus megaterium at MICs of 0.2 and 0.05 mM¹⁷.

Recent investigations with phytol demonstrated anxiolytic, metabolism-modulating, cytotoxic, antioxidant, autophagy and apoptosis-inducing, antinociceptive, anti-inflammatory, immune-modulating and antimicrobial effects¹⁸. The beta-sitosterol was found to possess anti-inflammatory, anticancer, antioxidant, hepatoprotective, cardioactive and antidiabetic¹⁹. The 1,2,3-benzenetriol (Pyrogallol) has the ability to perform antibacterial and antioxidant activities²⁰. The ethyl-alpha-d-glucopyranoside, skin moisturing agent²¹. The consumption of a food or beverage enriched with sucrose has been associated with improve mental alertness, memory, reaction time, as well as a reduction in the feeling of fatigue²². Oleic acid was found to have antibacterial activity, particularly in inhibiting the growth of several Gram-positive bacterial species²³.

CONCLUSION

The ethanolic extracts from Guiera senegalensis and Geigeria alata has revealed a wealth of diverse bioactive compounds, identified and quantified through Gas Chromatography-Mass Spectrometry (GC/MS). Guiera senegalensis displayed a significant presence of sugars, alcohols and ketones, suggesting its potential for therapeutic applications. Conversely, Geigeria alata exhibited a distinct chemical profile rich in ketones, fatty acids, alcohols and steroidal compounds, hinting at its potential for various physiological activities. These findings underscore the importance of botanical exploration in the search for new therapeutic agents and highlight the chemodiversity inherent in Sudanese plant species, thus offering valuable insights into the potential utilization of these botanical resources in drug discovery and development efforts.

SIGNIFICANCE STATEMENT

This study aimed to explore the compounds present in the ethanolic extracts of two important Sudanese plant species namely, Guiera senegalensis and Geigeria alata by implicating Gas Chromatography-Mass Spectroscopy (GC/MS) technique. The present study revealed that the ketonic, alcoholic, fatty acids and sugar derivatives compounds in studied two plants may be explored for manufacturing industrial products.

ACKNOWLEDGMENT

The authors thank Central Laboratory, Chemistry Department, University of Science and Technology, Ministry of Science and Technology, Khartoum, for the technical assistance.

REFERENCES

1. Elmalik, A.A., F.E.R. Ahmed and A. Abed Elsalam, 2022. Phytochemical analysis and antimicrobial activity of Guiera senegalensis leaves extraction. Scholars Int. J. Chem. Mater. Sci., 5: 118-121.
2. Kankara, S.S. and R. Go, 2016. Influence of extraction solvent on antioxidant properties of Guiera senegalensis J.F. Gmel (Combretaceae) leaves. Niger. J. Basic Appl. Sci., 24: 116-126.
3. Bosisio, E., D. Mascetti, L. Verotta, F. Zani, P. Mazza and M. Talbot, 1997. Guiera senegalensis J. F. Gmelin (Combretaceae): Biological activities and chemical investigation. Phytomedicine, 3: 339-348.
4. Silva, O. and E.T. Gomes, 2003. Guieranone A, a naphthyl butenone from the leaves of Guiera senegalensis with antifungal activity. J. Nat. Prod., 66: 447-449.
5. Moreira, R., F. Ferreres, Á. Gil-Izquierdo, N.G.M. Gomes and L. Araújo et al., 2023. Antifungal activity of Guiera senegalensis: From the chemical composition to the mitochondrial toxic effects and tyrosinase inhibition. Antibiotics, 12.
6. Fiot, J., S. Sanon, N. Azas, V. Mahiou and O. Jansen et al., 2006. Phytochemical and pharmacological study of roots and leaves of Guiera senegalensis J.F. Gmel (Combretaceae). J. Ethnopharmacol., 106: 173-178.
7. Mohammed, S.I.H., H.H. EL-Kamali and A.S. EL-Tahir, 2022. Haematological effects of aqueous extracts of Geigeria alata in male albino rats. J. Fac. Sci. Technol., 9: 125-133.
8. El-Kamali, H.H., 2009. Ethnopharmacology of medicinal plants used in North Kordofan (Western Sudan). Ethnobotanical Leafl., 13: 203-210.
9. Simeonova, R., V. Vitcheva, D. Zheleva-Dimitrova, V. Balabanova and I. Savov et al., 2019. Trans-3,5-dicaffeoylquinic acid from Geigeria alata Benth. & Hook.f. ex Oliv. & Hiern with beneficial effects on experimental diabetes in animal model of essential hypertension. Food Chem. Toxicol., 132.
10. Elegami, A.A., G.E.B. El Ghazali, W.E. Abdalla, G. Pintore, M. Chessa and B. Tirillini, 2006. Essential oil composition of Geigeria alata (DC.) Oliv. et Hiern. from Sudan. J. Essent. Oil Res., 18: 448-450.
11. Ross, S.A., K.A. El Sayed, M.A. El Sohly, M.T. Hamann, O.B. Abdel-Halim, A.F. Ahmed and M.M. Ahmed, 1997. Phytochemical analysis of Geigeria alata and Francoeuria crispa essential oils. Planta Med., 63: 479-482.
12. Fadul, E., A. Nizamani, S. Rasheed, A. Adhikari and S. Yousuf et al., 2020. Anti-glycating and anti-oxidant compounds from traditionally used anti-diabetic plant Geigeria alata (DC) Oliv. & Hiern. Nat. Prod. Res., 34: 2456-2464.
13. Zheleva-Dimitrova, D., R. Gevrenova, M.M. Zaharieva, H. Najdenski and S. Ruseva et al., 2017. HPLC‐UV and LC-MS analyses of acylquinic acids in Geigeria alata (DC) Oliv. and Hiern. and their contribution to antioxidant and antimicrobial capacity. Phytochem. Anal., 28: 176-184.
14. Vitcheva, V.B., R.L. Simeonova, M.S. Kondeva-Burdina, D.Z. Zheleva-Dimitrova, V.I. Balabanova, S. Yagi and R.S. Gevrenova, 2018. In vivo evaluation of the antioxidant potential of dicaffeoylquinic acid isolated from Geigeria alata. Bulg. Chem. Commun., 50: 98-104.
15. Siswadi and G.S. Saragih, 2021. Phytochemical analysis of bioactive compounds in ethanolic extract of Sterculia quadrifida R.Br. AIP Conf. Proc., 2353.
16. Sirikhansaeng, P., T. Tanee, R. Sudmoon and A. Chaveerach, 2017. Major phytochemical as γ-sitosterol disclosing and toxicity testing in lagerstroemia species. Evidence-Based Complementary Altern. Med., 2017.
17. Dilika, F., P.D. Bremner and J.J.M. Meyer, 2000. Antibacterial activity of linoleic and oleic acids isolated from Helichrysum pedunculatum: A plant used during circumcision rites. Fitoterapia, 71: 450-452.
18. Islam, M.T., E.S. Ali, S.J. Uddin, S. Shaw and M.A. Islam et al., 2018. Phytol: A review of biomedical activities. Food Chem. Toxicol., 121: 82-94.
19. Khan, Z., N. Nath, A. Rauf, T.B. Emran and S. Mitra et al., 2022. Multifunctional roles and pharmacological potential of β-sitosterol: Emerging evidence toward clinical applications. Chem. Biol. Interact., 365.
20. Cynthia, F. Ignatia, H. Sutanto and A. Darmawan, 2018. Antibacterial and antioxidant activities of pyrogallol and synthetic pyrogallol dimer. Res. J. Chem. Environ., 22: 39-47.
21. Dai, J., Y. Hu, Q. Si, Y. Gu, Z. Xiao, Q. Ge and R. Sha, 2022. Antioxidant and hypoglycemic activity of sequentially extracted fractions from pingguoli pear fermentation broth and identification of bioactive compounds. Molecules, 27.
22. Navarro, S.Z. and F.P. Llamas, 2013. Importance of sucrose in cognitive functions knowledge and behavior. Nutrición Hospitalaria, 28: 106-111.
23. Sales-Campos, H., P.R. de Souza, B.C. Peghini, J.S. da Silva and C.R. Cardoso, 2013. An overview of the modulatory effects of oleic acid in health and disease. Mini-Rev. Med. Chem., 13: 201-210.