GC-MS Analysis of Phytocomponents In the Ethanolic Leaf Extract of Allium Fistulosum Linn

RESEARCH ARTICLE Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 Please cite this article as: Joby J et al., GC-MS Analysis of Phytocomponents In the Ethanolic Leaf Extract of Allium Fistulosum Linny. American Journal of PharmTech Research 2025. GC-MS Analysis of Phytocomponents In the Ethanolic Leaf Extract of Allium Fistulosum Linn G.N Pramodini, Jesvy Joby * 1 Dept. Of Pharmacognosy , Nehru College of Pharmacy Pampady, Thrissur – 680588 ABSTRACT The present study investigates the phytochemical composition of the ethanolic leaf extract of Allium fistulosum L. (Welsh onion) using Gas Chromatography–Mass Spectrometry (GC–MS). The analysis identified 38 bioactive compounds, including fatty acids, esters, sterols, alcohols, phenolics, and other secondary metabolites. Major constituents detected were n-hexadecanoic acid (27.81%), DL-Proline, 5-oxo-, ethyl ester (10.77%), γ-sitosterol (7.94%), 9, 12, 15octadecatrienoic acid (4.60%), and phytol (4.22%). These compounds are reported to exhibit various pharmacological activity such as antimicrobial, antioxidant, anti-inflammatory, and potential anticancer activities. The phytoconstituents identified highlight the therapeutic relevance of A. fistulosum and provide scientific support for its traditional medicinal applications. The findings further suggest potential utility of this plant in pharmaceutical, nutraceutical, and cosmetic formulations. Keywords: GC-MS study, Therapeutic, Phytocomponents, Chromatogram *Corresponding Author Email: [email protected]om Received 22 August 2025, Accepted 26 September 2025 Journal home page: http://www.ajptr.com/ Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 17 www.ajptr.com INTRODUCTION Medicinal plants are among the earliest forms of therapy, having been used in traditional medicine across different cultures worldwide for thousands of years. Over generations, communities have preserved and transmitted empirical knowledge of their therapeutic benefits. Many plant species have had their biological activities and bioactive constituents identified, while others remain underexplored or require further scientific investigation. Despite these gaps, medicinal plants continue to hold promising potential for future healthcare applications. [1] The Allium genus has a long history of use in traditional medicine. Modern research on Allium species now focuses on evaluating the pharmacological properties of their extracts and active compounds. [2] Among them, green onions (Allium fistulosum) commonly referred to as scallion, Welsh onion, Japanese bunching onion, or spring onion belong to the Amaryllidaceae family and are well known for their distinctive flavour and wide culinary applications. [3] Extracts derived from green onions have demonstrated various therapeutic potentials, including anti-inflammatory, antimicrobial, anticancer, and anti-arthritic effects. Their medicinal significance has been acknowledged for centuries, particularly within traditional healing practices. [4] Different plant parts such as bulbs, pseudostem juice, leaves, flowers, seeds, and roots are valued for multiple bioactivities, including antibacterial, antitumor, antihypertensive, anti-obesity, antioxidant, cardiovascular activation, antiplatelet aggregation, relief of intestinal spasms, and immune system regulation. The key bioactive constituents include volatile oils (mainly sulphides), oleic acid, linoleic acid, allicin, pectin, and vitamin C. [5] The present study aims to identify phytoconstituents in the ethanolic leaf extract of Allium fistulosum Linn. Using GC-MS analysis, to evaluate its medicinal potential. MATERIALS AND METHOD Plant Material Collection, Authentication The leaves of Allium fistulosum l., were collected on 27 January 2025, from Thrissur vegetable market, district of Kerala. The plant specimen had been taxonomically identified and authenticated by Dr. Ranjusha A. P., Head of the Department of Botany at N. S. S. College, Ottapalam. The leaves were separated from the stem, thoroughly washed and they were shade-dried for about 25 to 30 days, ground into powder using a mixer grinder and kept in an airtight storage unit for subsequent studies Preparations Of Plant Powder The entire Allium fistulosum plant was washed and allowed to dry in the shade for a period of 25 to 30 days. After being ground in a mixer to a coarse powder, the dried plant was sealed in an airtight Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 18 container. In the phytochemical research, extraction is the first step. It involves applying certain solvents and following established protocols to separate the parts of plants that have therapeutic value. Preparation Of Extract Using a Soxhlet apparatus, 25 g of the dried, coarsely ground plant material of Allium fistulosum L. was extracted with a high-polarity solvent (ethanol) at 60° to 70° for 18 hours, or until the solvent became colorless in the siphon tube. The extracts were concentrated using a rotary evaporator. The resulting semisolid residue was gathered and kept in desiccators. The extract was subjected to GCMS analysis. [6] Characterization by GC-MS The ethanol extract of Allium fistulosum leaves was subjected to gas chromatography–mass spectrometry (GC–MS) analysis at the Kerala Forest Research Institute (KFRI) on 23rd July 2025 to identify its phytochemical constituents. The GC–MS procedure can be carried out under the following conditions. The sample should first be clarified by filtration or centrifugation, followed by extraction of analytes with an appropriate solvent such as hexane or dichloromethane. An internal standard may be added, and the extract concentrated if necessary, before transferring to a clean auto sampler vial. A capillary GC column (30 m × 0.25 mm × 0.25 µm, nonpolar phase) is used, with helium as the carrier gas at a constant flow rate of 1.0 mL/min. The injection port temperature is set between 250–280 °C, and depending on analyte concentration, either split or splitless injection is employed, with 0.5–1.0 µL sample volume. The oven is programmed to start at 40 °C (held for 2 min), increase at 10 °C/min to 150 °C, then 5 °C/min to 250 °C, and further at 10 °C/min to 300 °C, with a final hold of 5–10 min, ensuring the total run includes the 7.00–51.00 min MS acquisition range. A solvent delay of 6.50 min is applied to protect the detector from solvent peaks. In the MS system, the ion source temperature is maintained at 230 °C, the interface at 280 °C, with electron ionization at 70 eV and a detector gain of 0.90 kV relative to tuning. Data are collected in scan mode across m/z 35–500 with an event time of 0.30 s and a scan speed of 1666. The analysis begins with a blank run to monitor background, followed by calibration standards, quality control injections, and then the sample runs, with QC checks included at intervals. Peaks are identified by comparing spectra with reference libraries such as NIST or Wiley, confirmed against standards when available, and quantified using calibration curves with the internal standard. Throughout the procedure, good laboratory practices are followed, including the use of PPE, handling solvents under a fume hood, safe waste disposal, and troubleshooting for sensitivity, peak shape, or carryover issues when required.[7] Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 19 www.ajptr.com RESULTS AND DISCUSSION Graph 1: Chromatogram of EEAF Table 1: GC-MS of EEAF Peak R. time Mol wt Formula Compound Name 1 7.619 144 C 6 H 8 O 4 4H - Pyran - 4 - one, 2,3 - dihydro - 3,5 - dihydroxy - 6 - methyl - 2 8.063 146 C 6 H 10 O 4 Ethyl hydrogen succinate 3 8.762 120 C 8 H 8 O 4 - Vinylphenol 4 9.250 136 C 8 H 8 O 2 Benzeneacetic acid 5 9.458 158 C 9 H 18 O 2 Nonanoic acid 6 10.078 150 C 9 H 10 O 2 2 - Methoxy - 4 - vinylphenol 7 12.024 157 C 7 H 11 NO 3 DL - Proline, 5 - oxo - , ethyl ester 8 13.022 168 C 9 H 12 O 3 Homovanillyl alcohol 9 13.342 200 C 12 H 24 O 2 Dodecanoic acid 10 14.390 No hit compound 11 15.456 226 C 13 H 22 O 3 7-Oxabicyclo[4.1.0] heptan-3-ol, 6-(3-hydroxy-1-butenyl)-1,5,5trimethyl - 12 15.675 228 C 14 H 28 O 2 Tetradecanoic acid 13 15.976 196 C 11 H 16 O 3 6-Hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydrobenzofuran2(4H) - one 14 16.669 278 C 20 H 38 Neophytadiene 15 17.355 278 C 20 H 38 Neophytadiene 16 18.156 270 C 17 H 34 O 2 Hexadecanoic acid, methyl ester 17 18.591 296 C 20 H 40 O Phytol 18 19.184 256 C 16 H 32 O 2 n - Hexadecanoic acid 19 19.615 284 C 18 H 36 O 2 Hexadecanoic acid, ethyl ester 20 21.000 268 C 17 H 32 O 2 cis - 10 - Heptadecenoic acid 21 21.322 270 C 17 H 34 O 2 Heptadecanoic acid 22 22.002 294 C 19 H 34 O 2 9,12 - Octadecadienoic acid ( Z, Z ) - , methyl ester 23 22.133 292 C 19 H 32 O 2 9,12,15 - Octadecatrienoic acid, methyl ester, ( Z, Z ,Z) - Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 20 24 22.415 296 C 20 H 40 O Phytol 25 23.266 278 C 18 H 30 O 2 9,12,15 - Octadecatrienoic acid, ( Z, Z ,Z) - 26 23.320 310 C 20 H 38 O 2 cis - 13 - Eicosenoic acid 27 23.524 308 C 20 H 36 O 2 Linoleic acid ethyl ester 28 23.667 352 C 21 H 36 O 4 Linolenic acid, 2 - hydroxy - 1 - (hydroxymethyl)ethyl ester ( Z, Z ,Z) - 29 24.263 312 C 20 H 40 O 2 Octadecanoic acid, ethyl ester 30 27.242 280 C 20 H 40 Cyclohexane, tetradecyl - 31 28.154 382 C 26 H 54 O 1 - Hexacosanol 32 29.888 274 C 16 H 31 ClO Palmitoyl chloride 33 30.647 390 C 24 H 38 O 4 Bis(2 - ethylhexyl) phthalate 34 36.975 402 C 27 H 46 O 2 delta. - Tocopherol 35 39.080 384 C 27 H 44 O Cholesta - 4,6 - dien - 3 - ol, (3.beta.) - 36 40.084 386 C 27 H 46 O Cholesterol 37 43.221 412 C 29 H 48 O Stigmasterol 38 44.976 414 C 29 H 50 O gamma. - Sitosterol Graph 2: Chromatogram of 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methylGraph 3: Chromatogram of Ethyl hydrogen succinate Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 21 www.ajptr.com Graph 4: Chromatogram of 4-Vinylphenol Graph 5: Chromatogram of Benzeneacetic acid Graph 6: Chromatogram of Nonanoic acid Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 22 Graph 7: Chromatogram of 2-Methoxy-4-vinylphenol Graph 8: Chromatogram of DL-Proline, 5-oxo-, ethyl ester Graph 9: Chromatogram of Homovanillyl alcohol Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 23 www.ajptr.com Graph 10: Chromatogram of Dodecanoic acid Graph 11: No hit compound Graph 12: Chromatogram of 7-Oxabicyclo[4.1.0]heptan-3-ol, 6-(3-hydroxy-1-butenyl)-1,5,5-trimethyl Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 24 Graph 13: Chromatogram of Tetradecanoic acid Graph 14: Chromatogram of 6-Hydroxy-4,4,7a-trimethyl-5,6,7,7atetrahydrobenzofuran-2(4H)-one Graph 15: Chromatogram of Neophytadiene Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 31 www.ajptr.com Graph 34: Chromatogram of Bis (2-ethylhexyl) phthalate Graph 35: Chromatogram of delta. -Tocopherol Graph 36: Chromatogram of Cholesta-4, 6-dien-3-ol, (3. beta.)- Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 32 Graph 37: Chromatogram of Cholesterol Graph 38: Chromatogram of Stigmasterol Graph 39: Chromatogram of gamma. -Sitosterol Joby et. al., Am. J. PharmTech Res. 2025; 15(05) ISSN: 2249-3387 33 www.ajptr.com The GC–MS analysis revealed a diverse phytochemical profile consisting of fatty acids, fatty acid esters, flavonoid, phenolic compounds, alcohols, sterols, and other bioactive metabolites. Major constituents identified include n-hexadecanoic acid (27.81%), DL-Proline, 5-oxo-, ethyl ester (10.77%), γ-sitosterol (7.94%), 9,12,15-octadecatrienoic acid (4.60%), and phytol (4.22%), which are known for their antimicrobial, antioxidant, anti-inflammatory, and potential therapeutic activities. The high proportion of saturated and unsaturated fatty acids, along with phytosterols and tocopherols, indicates significant nutritional and pharmacological potential. This compositional profile supports the traditional uses of the sample and suggests promising applications in food, cosmetic, and pharmaceutical formulations. CONCLUSION The GC–MS analysis of ethanolic extracts of Allium fistulosum leaves revealed a rich phytochemical profile comprising fatty acids, phenolic derivatives, phytosterols, tocopherols, and other biologically active compounds. The predominance of n-hexadecanoic acid, DL-Proline derivatives, and γ-sitosterol indicates strong pharmacological potential, particularly in antimicrobial, antioxidant, and anti-inflammatory applications. These results validate the traditional medicinal uses of A. fistulosum and emphasize its importance as a natural source of therapeutic agents. Further studies focusing on isolation, biological evaluation, and formulation development are recommended to fully exploit its medicinal and commercial potential. ACKNOWLEDGMENT I would like to thank Nehru College of pharmacy for the support and lab facilities. REFERENCE 1. Marrelli M. Medicinal Plants. Plants [Internet]. 2021 Jul 1; 10(7):1355. 2. Maragheh FP, Janus D, Senderowicz M, Haliloglu K, Kolano B. Karyotype analysis of eight cultivated Allium species. Journal of applied genetics. 2019 Feb 7; 60:1-1. 3. Shinde V, Dhalwal K. Pharmacognosy: The changing scenario. Pharmacognosy Reviews. 2007 Jan 1; 1(1):1-6. 4. Kim SH, Yoon JB, Han J, Seo YA, Kang BH, Lee J, Ochar K. Green onion (Allium fistulosum): an aromatic vegetable crop esteemed for food, nutritional and therapeutic significance. Foods. 2023 Dec 16; 12(24):4503. 5. Padula G, Xia X, Hołubowicz R. Welsh onion (Allium fistulosum L.) seed physiology, breeding, production and trade. Plants. 2022 Jan 27; 11(3):343. Joby et. al., Am. J. PharmTech Res. 2025;15(05) ISSN: 2249-3387 www.ajptr.com 34 6. Monika N, and Sakthi A. Evaluation of invitro anticancer activity of bulb extracts of allium fistulosum l. against hepatocellular carcinoma cell line (hep g2). World journal of pharmacy and pharmaceutical sciences. 2018 Apr 30; Volume 7(Issue 5):1678–99. 7. Monika N, Sakthi Abirami M. GC-MS studies on traditional plant Allium fistulosum L. bulbs. World Journal of Pharmacy and Pharmaceutical Sciences. 2018; 7(6):823-834. AJPTR is  Peer-reviewed  bimonthly  Rapid publication Submit your manuscript at: edito[email protected]