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INTRODUCTION
The plant Syzygium samarangense was traditionally used as a best medicine for many systemic diseases. Several researchers were documented the potential benefits of many phytoconstituents of this plant including flavonoids and anthocyanins. The pharmacological effect of various extracts of this plant already documented well also (Khamchan et al., 2018; Mahmoud et al., 2021; Sobeh et al., 2018). Myricitrin and 3,5-di-O-methyl gossypetin were isolated from the leaf of Syzygium samarangense and the anti-oxidative effect of these components were well documented (Sobeh et al., 2019). Several components of methanolic leaf extract of Syzygium samarangense were isolated. Most of the components belongs to flavonoid, tannins and polyphenols (Sobeh et al., 2018). Many older studies reported that, the plant Syzygium samarangense contains phenolic components as secondary metabolites with promising pharmacological activities (Hassan et al., 2022).
Gas Chromatography coupled on-line with Mass Spectrometry (GC-MS) is one of the best instrumental method to determine the secondary metabolites presented in the plant extracts. The characterization of phytoconstituents through GC-MS can provide complete metabolite profile (Frolova et al., 2026). The GC-MS methods is modern technique, which can separate and identify the phytoconstituents in single step by using GC-MS library (Arifuzzaman et al., 2025). Hence, the present study was designed to investigate the important phytoconstituents present in the aqueous extract of root of Syzygium samarangense by using GC-MS method.
MATERIALS AND METHODS
Plant Collection and Authentication
The root of Syzygium samarangense was collected from surrounding village of Rajampet Kadapa, A.P, India in the month of September 2018. The root was further verified and authenticated as the root of Syzygium samarangense (Blume) Merr and L.M. Perry from the family of Myrtaceae by Dr. M.V. Suresh Babu Department of Botany, Govt Degree College, Rajampet, Kadapa, A.P, India and voucher specimen number was lodged (ANCP-MP-COL-01/2018-19).
Preparation of Aqueous extract of root of Syzygium samarangense
Fresh roots were collected and dried in the shade. The shade-dried roots of the plant (500 g) were powdered, sieved through a mesh of size 80 to obtain a powder of uniform size, soaked in cold water (cold percolation) for 3 days, and the aqueous extract were decanted. The solvent of the total liquid extract was evaporated by distillation to a concentrate over a water bath to a syrupy consistency and then evaporated to dryness under vacuum to give the dry extract (16%w/w yield). The extract was stored at 4ºC for further analysis.
Phytochemical Screening
The crude aqueous extract subjected to preliminary phytochemical screening to identify the major phytochemical constituent’s by using standard protocols. The Total Phenolic Content (TPC) and Total Flavonoid Content (TFC) were analysed by using the Folin-Ciocalteu reagent method with gallic acid as the standard and aluminum chloride colorimetric method with quercetin as the standard respectively, following a standard protocol with minor modifications (Arifuzzaman et al., 2025).
TPC determination
2 mg of the extract were dissolved in distilled water to obtain a stock concentration of 1 mg/mL. An aliquot of 0.5 mL of this extract was mixed with 2 mL of diluted Folin-Ciocalteu reagent and allowed to stand at 22 ± 2ºC for 5 min. Subsequently, 2.5 mL of 7.5% (w/v) sodium carbonate solution was added, and the mixture was gently vortexed and incubated for 20 min for color development. The absorbance was measured at 760 nm using a UV-visible spectrophotometer. TPC was calculated from the gallic acid calibration curve and expressed as mg Gallic Acid Equivalents (GAE) per g of dry extract.
TFC determination
Briefly, 1.5 mL of the extract was mixed with 0.1 mL of 10% AlCl₃, followed by 0.1 mL of 1 M sodium acetate. After standing for 30 min, 1 mL of 1 M NaOH was added, and the volume was adjusted to 5 mL with double-distilled water. The reaction mixture was incubated for 15 min, and absorbance was recorded at 415 nm. TFC was calculated from the quercetin calibration curve and expressed as mg Quercetin Equivalents (QE) per gram of dry extract.
The TLC profiling was analysed as described by a previous study by using silica gel plates and the mobile phase n-hexane:ethyl acetate: glacial acetic acid (85:20:5 v/v/v). The extract was spotted onto the plate and developed in a saturated chamber until the solvent front reached an appropriate height. After drying, the chromatogram was visualized under UV light (Appamaraka et al., 2022).
GC-MS Analysis
The GC-MS analysis was carried out with 6890 N Agilent gas chromatograph attached with a JMS 600 H JEOL mass spectrometer. The phyto constituents present in the extract was separated on a fused capillary SPBI column, 30 m 0.32 mm, 0.25 μm film thicknesses in a temperature program initially 50 to 256ºC with a rate of 4ºC/min with 2 min hold. The injector was at 260ºC and the flow speed of the carrier gas helium, was 1 mL/min. The EI mode JMS 600 H JEOL mass spectrometer had ionization volt of 70 Ev, electron emission of 100 Μa, ion source temperature of 250ºC and analyzer temperature of 250°C. The extract was injected 1 µL manually in split mode with the ratio of sample in split mode was 20:1. GC-MS detection was based on the computer evaluation of mass spectra of samples through National Institute Standard and Technology (NIST), through comparison of peaks and retention time and computer matching as well as by following the characteristic fragmentation patterns of the mass spectra of particular class of compounds.
Statistical analysis
The research work is purely depending on phytochemical analysis of aqueous extract of root of Syzygium samarangense. Hence, only description of GC-MS results is included without any specific descriptive statistical analysis
RESULTS
Preliminary phytochemical investigation revealed the presence of terpenoids, carbohydrates, lipids, and flavonoids. Quantitative analyses revealed a substantial Total Phenolic Content (TPC) ranging from 61.41 to 143.68 ppm and Total Flavonoid Content (TFC) between 45.76 and 148.53 ppm in the aqueous extract of the root. TLC analysis of the extract confirmed that the mobile phase n-hexane: ethyl acetate: glacial acetic acid in the ratio of 85:20:5 gave better separation, and three compounds were observed with Rf values of 0.36, 0.82, and 0.94 Figure 1.
The aqueous extract of root of Syzygium samarangense was analyzed by GC-MS to detect various compounds with the help of NIST library. Totally 11 major compounds were identified which have been listed in Table 1. The chromatograph showed 48 peaks with 48 individual compounds Figure 2.
| Sl. No. | Retention Time | Peak area % | Name of the compound | Molecular formula | Molecular weight | Chemical structure | Nature of the compound |
|---|---|---|---|---|---|---|---|
| 1 | 7.045 | 2.796 | 1,4-Dioxane-2,5-Dione, 3,6- Dimethyl | C6H8O4 | 144 | Lactone derived from lactic acid | |
| 2 | 18.36 | 37.943 | Beta-D-Mannofuranoside, Methyl | C7H14O6 | 194 | Carbohydrate Derivative Acts as A Substrate for glucosidase | |
| 3 | 19.61 | 13.547 | N-Hexadecanoic Acid | C16H32O2 | 256 | Saturated Fatty acid known as Palmitic acid | |
| 4 | 19.885 | 7.054 | Octadecanoic acid | C18H36O2 | 284 | Saturated fatty acid known as Stearic acid | |
| 5 | 20.265 | 1.441 | 1-O-Acetyl-Exo-2,3-O-Ethylidine-Beta-D-Erythrofuranose | C8H12O5 | 188 | Cyclic hemiacetal Carbohydrate Derivative | |
| 6 | 20.371 | 3.077 | Hexadecanoic Acid,1,1-Dimethylethyl Ester | C20H40O2 | 312 | Ester of Palmitic acid | |
| 7 | 20.896 | 23.139 | 9-Oxanonanoic Acid | C9H16O3 | 172 | Medium-chain oxo-fatty acid, acetyl-CoA carboxylase inhibitor | |
| 8 | 21.656 | 2.095 | 11-Tridecen-1-Ol | C13H26O | 198 | Fatty alcohol | |
| 9 | 21.746 | 1.598 | Hexadecanoic Acid,2-Hydroxy-Methyl Ester, Acetate | C19H36O4 | 328 | Esterified form of Palmitic acid | |
| 10 | 21.846 | 2.133 | T-Butyl Cyclopentane peroxy carboxylate | C10H18O3 | 186 | Hydrocarbon | |
| 11 | 23.257 | 5.179 | Di-N-Octyl phthalate | C24H38O4 | 390 | Esters of phthalic acid |
DISCUSSION
GC-MS profiling of the Syzygium samarangense root extract revealed a diverse array of bioactive metabolites, predominantly therapeutic fatty acids, phytosterols, and terpenoids. These findings are consistent with comprehensive phytochemical reviews of the species, which report sterols among the major classes of metabolites found in Syzygium samarangense (Sandhiya and Amudha, 2025; Tarigan, 2021). The synergistic action of compounds such as n-hexadecanoic acid, squalene, lupeol, and eugenol imparts significant antimicrobial, anti-inflammatory, and antioxidant properties to the root extract. These findings strongly validate the traditional ethnomedicinal applications of the plant roots and underscore their potential as a viable source of novel plant-based pharmacological therapeutics. Ultimately, this study advances the pharmacognostic understanding of Syzygium samarangense and supports the sustainable utilization of medicinal plant resources for human health benefits.
CONCLUSION
The findings from the present analysis indicate that Syzygium samarangense root extract possesses a diverse array of bioactive phytoconstituents that collectively support its traditional medicinal applications and pharmacological potential.