Phytochemical Profiling and GC-MS Analysis of Conocarpus lancifolius Fruits

Received 30 Oct, 2025

Accepted 24 Jan, 2026

Published 31 Mar, 2026

Background and Objective: Plants are repositories of several natural chemicals; the identification of which are important in the field of medicine, cosmetics, health and other industries. Conocarpus lancifoliusEngl.(Family-Combretaceae) is an evergreen tree cultivated as an ornamental plant species. Since no comprehensive phytochemical analysis of its fruits was available, an attempt was made to evaluate the phytochemical composition and phyto-constituents present in its methanolic fruit extract. Materials and Methods: Both dry (mature) and green (immature) fruits (Dfr and Gfr, respectively) were collected from Udaipur city, Rajasthan, India. The aqueous and methanolic extracts underwent preliminary qualitative phytochemical screening, while the methanolic extract was subjected to Gas Chromatography-Mass Spectrometry (GC-MS) analysis for the first time. Results: The preliminary screening revealed the presence of carbohydrates, flavonoids, phenols, cardiac glycosides, terpenoids, coumarins, tannins, steroids, and phlobatannins, and the absence of amino acids and saponins. The GC-MS analysis demonstrated the presence of 80 and 65 biological compounds in Dfr and Gfr, respectively. The major compounds identified in Dfr and Gfr were, 1,2,3-benzenetriol (13.19, 26.13%), n-hexadecanoic acid (15.92, 11.12%), β-sitosterol (10.84, 9.57%), 10(E),12(Z)-Conjugated linoleic acid (9.51, 2.66%) and Lupeol (5.09, 5.57%), respectively. Conclusion: The preliminary phytochemical analysis and GC-MS profiling of C. lancifolius fruits from India have shown the presence of biologically significant phytochemicals for the first time and could stimulate future research to evaluate its complete pharmacological potential.

INTRODUCTION

Metabolites are small molecules produced during metabolic processes and can be classified as primary or secondary. Primary metabolites, such as vitamins, amino acids, nucleotides, and organic acids, are essential for growth, reproduction, and normal physiological functions. In contrast, secondary metabolites, synthesized mainly during the stationary growth phase, are not directly involved in growth but play vital roles in plant defense and adaptation found abundantly in medicinal plants¹. Moreover, the secondary metabolites, for example, alkaloids, flavonoids, tannins, phenolics, and saponins, exhibit diverse biological activities with significant therapeutic value².

Fig. 1: Conocarpus lancifolius Engl

A qualitative and quantitative analysis of the phyto-constituents offers valuable insights for the discovery of new drugs. Gas Chromatography-Mass Spectrometry (GC-MS) is a reliable analytical method that has been widely used for the structural elucidation of a broad range of phyto-constituents, such as glycosides, flavonoids, phenolics, essential oils, alkaloids, saponins, steroids, and their derivatives, in addition to the analysis of volatile compounds³. Thus, the GC-MS profiling of a plant extract helps in the identification of pharmacologically valuable compounds.

Conocarpus lancifolius Engl. (Fig. 1) belonging to the family Combretaceae is an ornamental evergreen tree native to Somalia. It is commonly known as Lanceleaf Buttonwood and Damas tree and is notable for its resistance to heat and salt, as well as its tolerance to drought⁴. The plant is under the vulnerable category as per the IUCN Red Data List, and its decreasing population trend is a matter of concern⁵. Aerial parts, leaves, roots, and fruits of this plant have been explored for their phyto-pharmaceutical potential, and several pharmacological activities, including antioxidant, antimicrobial, anxiolytic, antidiabetic, cytotoxic, antiurease, cardio-protective, antiquorum-sensing, and acetylcholinesterase inhibition have been reported^6-14. Besides, many secondary metabolites like phenolic compounds, flavonoids, alkaloids, fatty acids, steroids, terpenoids, coumarins, saponins, tannins, glycosides, anthraquinones etc. have been identified from leaves, roots, stems, and aerial parts of C. lancifolius.

Al-Taweel et al.⁶ have isolated one novel compound 3,3’4' trimethoxy 4-O-cyclopentanone ellagic acid, and two known compounds, kaempferol 3-O-rutinose and β sitosterol 3-O-glucoside, from the fruits of C. lancifolius. Whereas Afifi et al.⁷ have reported several polyphenolic compounds from ethanolic fruit extract of C. lancifolius, such as 4-hydroxy benzoic acid, vanillic acid, caffeic acid, 1,2-dihydroxy benzene, catechin, benzoic acid, p-coumaric acid, t-ferulic acid, sinapic acid, vanillin, rutin hydrate, cinnamic acid, protocatechuic acid, quercetin, and tannins. However, phytochemical investigationofto its fruits through GC-MS have still not been carried out. Hence, the present study was undertaken to investigate the bioactive compounds present in the methanolic extract of Conocarpus lancifolius fruits, contributing to a better understanding of its phytochemical composition.

MATERIALS AND METHODS

Plant collection and identification: The dry (fully mature) and green (immature) fruits of C. lancifolius were collected during February-March, 2025 from Udaipur City, Rajasthan, India. The voucher specimen was prepared and authentication of the plant was carried out at Arid Zone Regional Centre, Botanical Survey of India (BSI), Jodhpur (No./BSI/AZRC/A.12012/Tech./2025-26(Pl. Id.)/445; dated 17.09.2025).

Preparation of plant extracts: Both dry and green fruits were washed under running tap water and air-dried in the shade at room temperature. After drying, both were powdered separately, coded as Dfr and Gfr, respectively, and stored at 4°C in a refrigerator. For preliminary qualitative phytochemical and GC-MS analyses, the following two types of extracts were prepared from the powdered plant materials.

Aqueous extract: Aqueous extracts were freshly prepared by soaking 400 mg of each Dfr and Gfr of C. lancifolius in 20 mL of distilled water separately, followed by boiling for 20 min. These were cooled and filtered through Whatman’s No. 1 filter paper. The clear filtrates were utilized instantly for preliminary qualitative phytochemical screening.

Methanolic extract: One hundred gram of both Dfr and Gfr were soaked in 700 mL of methanol separately for eight days with periodic stirring and then filtered using Whatman’s filter paper No. 1. The filtrates were evaporated in a boiling water bath at 40°C, and the concentrated extracts were stored in sterile glass petri dishes at 4ºC in the refrigerator. The percent yields obtained for methanolic extracts of Dfr and Gfr were 1.88 and 10.34%, respectively. These extracts were used for qualitative phytochemical assessment as well as analyzed for the presence of various phytochemicals through GC-MS.

Qualitative phytochemical screening: Aqueous/ME or dry and green fruit powders of C. lancifolius were used for preliminary qualitative phytochemical screening of amino acids, carbohydrates, terpenoids, steroids, cardiac glycosides, phlobatannins, flavonoids, phenols, tannins, coumarins, and saponins as required. These tests were performed as per standard methodology¹⁵.

Gas Chromatography-Mass Spectrometry (GC-MS) analysis: To find out the chemical compounds present in the methanolic extracts of Dfr and Gfr of C. lancifolius, Gas Chromatography-Mass Spectrometry analysis was conducted on a Shimadzu GC-MS-QP2010 Ultra system with an AOC-20i+s autosampler. The system employed a Rxi-5 SIL MS column (30 m×0.25 mm i.d×0.25 mm film thickness) with helium as the carrier gas at a constant flow rate of 1.21 mL/min. In split mode, the injector operated with a 10:1 split ratio, 1 μL of injection volume, and 270°C injection temperature. The column oven temperature was initially established at 70°C for 2 min, subsequently increased at a rate of 10°C/min to 300°C, and held for 15 min. Ion source and interface temperatures were maintained at 220°C and 280°C, respectively. In scan mode, mass spectra were obtained with a solvent cut-off time of 3.50 min, a mass range of 40-600 m/z, and a scan rate of 3333 scans per second. Data were acquired in Total Ion Count (TIC) mode. Identifications of compounds were made by matching retention times and mass spectral patterns against the National Institute of Standards and Technology (NIST) and Wiley libraries. The relative abundance of each analyte was computed as the percentage of its peak area relative to the aggregate peak area of all detected components.

RESULTS

Qualitative phytochemical analysis: A preliminary qualitative phytochemical analysis of both dry and green fruits of C. lancifolius has shown the presence of one primary metabolite that is carbohydrate and eight secondary metabolites, namely, flavonoid, phenol, cardiac glycoside, terpenoid, coumarin, steroid, tannin and phlobatanin. One primary metabolite amino acid and one secondary metabolite saponin were not detected in both Dfr and Gfr (Table 1).

GC-MS analysis: The methanolic extracts of dry and green fruits of C. lancifolius, analyzed through GC-MS, revealed 84 and 71 peaks in the chromatograms, corresponding to 80 and 65 compounds, respectively (Fig. 2 and 3). Among which, n-hexadecanoic acid (15.92%), pyrogallol (13.19%), β-sitosterol (10.84%), 10(E),12(Z)-Conjugated linoleic acid (9.51%), Oleic acid (5.36%) and Lupeol (5.09%) in Dfr (Table 2) whereas pyrogallol (26.13%), n-hexadecanoic acid (11.12%), β-sitosterol (9.57%) and Lupeol (5.57%) in Gfr (Table 3), were the most abundant compounds. Among the variety of secondary metabolites in C. lancifolius, terpenoids and fatty acid derivatives are predominant compounds present, followed by phenolic compounds, phytosterols, hydrocarbons, heterocyclic compounds, and other miscellaneous compounds (Table 4).

Fig. 2: GC-MS chromatogram of methanolic extract of Conocarpus lancifolius dry fruits

Fig. 3: GC-MS chromatogram of methanolic green fruit extract of Conocarpus lancifolius

The compounds which are unique to dry mature fruits of C. lancifolius are Diglycolic acid, ethyl 2-isopropoxyphenyl ester, Octanoic acid, Phenol, 8-Hydroxy-2-octanone, 2,4,6-Cycloheptatrien-1-one, Dehydromevalonic lactone, Dianhydromannitol, 3-Dimethylsilyloxytetradecane, Nonanoic acid, Acetic acid, 1,3,7-trimethylocta-2,6-dienyl ester, Dodecanal dimethyl acetal, 3-Methyl-2-(2-methyl-2-butenyl)- furan, 2-Isopropenyl-3-methylpyrazine, Guaia-6,9-diene, Longifolene, 6-Dimethyl(trimethylsilyl)silyloxytetradecane, D-Mannoheptulose, γ-Elemene, α-Amorphene, Dodecanoic acid, Canophyllal, Orcinol, monoacetate, Decane, 2,3,8-trimethyl-, Isopropyl palmitate, 2-Methyl tetracosane, Heptadecanoic acid, Oleic Acid, Undec-10-ynoic acid, tetradecyl ester, Eicosanoic acid, 1H-1,3-Benzimidazol-7-amine, N-[(4-methoxyphenyl)methyl, Tyrosine, 9-Hexacosene, 1-Decyloxymethyl-3-methyl-1,3-dihydrobenzoimidazol-2-ylideneamine, Ethyl 6,9,12,15-octadecatetraenoate, 9,12-Octadecadienoic acid (Z,Z)-, 2,3-dihydroxypropyl ester, Octadecanoic acid, 2,3-dihydroxypropyl ester, Benzoic acid, 2,4-dihydroxy-, 4-(1,1-dimethylpropyl)phenyl, Oleamide, 1,54-dibromotetrapentacontane, Dodecane, 1,1-dimethoxy-, Tetracosan-10-yl acetate, Campesterol, Stigmast-5-en-3-ol, oleat, Stigmasta-4,22-diene, Stigmastane-3,6-dione, (5-alpha), α-amyrin and Cyclopentanone, 2-(5-oxohexyl) as shown in Table 2. Besides, two peaks were obtained for four compounds, namely, 9-Octadecenamide, Phenol, 2-methyl-5-(1-methylethyl), Neophytadiene, and 10(E),12(Z)-Conjugated linoleic acid (Table 2).

Table 1:

Qualitative phytochemical analysis of fruits of Conocarpus lancifolius Engl.

Phytochemical	Dry fruit (Dfr)	Green fruit (Gfr)
Carbohydrate	+	+
Amino acid	-	-
Flavonoid	+	+
Phenol	+	+
Cardiac glycoside	+	+
Terpenoid	+	+
Coumarin	+	+
Steroid	+	+
Saponin	-	-
Tannin	+	+
Phlobatanin	+	+
+: Present and -: Absent

Table 2:

Phyto-compounds identified through GC-MS in methanolic extract of Conocarpus lancifolius dry fruits (Dfr)

Peak	R. Time	Area %	Name of the compound	Molecular formula	Molecular weight (g/mol)	Name of compound
1	4.549	0.05	Diglycolic acid, ethyl 2-isopropoxyphenyl ester	C15H20O6	296	Aromatic ester
2	4.616	0.06	Octanoic acid (Caprylic acid)	C8H16O2	144	Saturated fatty acid
3	4.763	0.15	Phenol	C6H5OH	94	Phenolic compound
4	5.9	0.01	4-hydroxy-2,5-dimethyl- 3(2h)-furanone	C6H8O3	128	Heterocyclic compound
5	6.257	0.07	8-Hydroxy-2-octanone	C8H16O2	144	Aliphatic hydroxyl ketone
6	7.362	0.23	4H-Pyran-4-one, 2,3-dihydro-3, 5-dihydroxy-6-methyl (Pyranone)	C6H8O4	144	Phenolic compound
7	7.508	0.02	2,4,6-Cycloheptatrien-1- one (Tropone)	C7H6O	106	Conjugated ketone
8	7.596	0.13	Dehydromevalonic lactone	C6H8O2	112	Cyclic α,β-unsaturated lactone
9	8.422	0.16	Dianhydromannitol	C6H10O4	146	Sugar alcohol derivative
10	8.762	0.06	3-Dimethylsilyloxytetradecane	C16H36OSi	272	Alkane derivative
11	9.107	0.18	Nonanoic acid	C9H18O2	158	Fatty Acid
12	9.185	0.03	Acetic acid, 1,3,7-trimethylocta- 2,6-dienyl ester	C13H22O2	210	Monoterpene ester
13	9.262	0.02	Dodecanal dimethyl acetal	C14H30O2	230	Fatty acid
14	9.333	0.08	Phenol, 2-methyl-5-(1-methylethyl), (O-Thymol/Carvacrol)	C10H14O	150	Phenolic compound
15	9.531	0.37	3-Methyl-2-(2-methyl-2-butenyl)- furan (Rosefuran)	C10H14O	150	Terpenoid
16	9.662	0.16	Phenol, 2-methyl-5-(1-methylethyl), (O-Thymol/Carvacrol)	C10H14O	150	Phenolic compound
17	10.289	0.02	2-Isopropenyl-3-methylpyrazine	C8H10N2	134	Heterocyclic compound
18	10.423	0.05	Phenol, 2-methoxy-4-(2-propenyl, (1,3,4-Eugenol)	C10H12O2	64	Phenolic compound
19	10.478	0.07	n-Decanoic acid (Capric acid )	C10H20O2	172	Fatty Acid
20	10.619	0.04	4-tert-Butylcyclohexyl acetate	C12H22O2	198	Acid ester
21	10.715	13.19	1,2,3-benzenetriol (Pyrogallol)	C6H3(OH)3	126	Phenolic compound
21	11.524	0.13	Guaia-6,9-diene	C15H24	204	Sesquiterpene
23	11.863	0.07	Longifolene	C15H24	204	Sesquiterpene
24	12.006	0.07	6-Dimethyl(trimethylsilyl)silyloxytetradecane	C19H44OSi2	344	Organosilane
25	12.248	0.28	D-Mannoheptulose	C7H14O7	210	Monosaccharide
26	12.46	0.07	γ-Elemene	C15H24	204	Sesquiterpene
27	12.64	0.15	α-Amorphene	C15H24	204	Sesquiterpene
28	13.019	0.38	Dodecanoic acid (Lauric acid)	C12H24O2	200	Fatty Acid
29	14.124	0.37	Isocitronellol	C10H20O	156	Monoterpenoid
30	14.577	0.23	Caryophyllane, 4,8- beta-epoxy	C15H26O	222	Sesquiterpene
31	14.98	0.29	Canophyllal	C30H48O2	440	Pentacyclic triterpenoid
32	15.309	0.48	Tetradecanoic acid (Myristic acid)	C14H28O2	228	Fatty Acid
33	15.712	0.1	Orcinol, monoacetate	C7H8O2	124	Phenolic compounds
34	15.778	0.21	Decane, 2,3,8-trimethyl-	C13H28	184	Branched alkane
35	16.007	0.17	Isopropyl palmitate	C19H38O2	298	Fatty acid ester
36	16.164	0.67	Neophytadiene	C20H38	278	Diterpene
37	16.216	0.33	2L, 4d-dihydroxyeicosane	C20H42O2	314	Fatty alcohol
38	16.616	0.4	Phytol (3,7,11,15-Tetramethyl- 2-hexadecen-1-ol)	C20H42O	296	Diterpenoid
39	17.062	1.47	Hexadecanoic acid, methyl ester (Palmitic acid methyl ester)	C17H34O2	270	Fatty acid ester
40	17.424	15.92	n-Hexadecanoic acid (Palmitic acid)	C16H32O2	256	Fatty acid ester
40	17.819	0.09	2-Methyl tetracosane	C25H52	352	Alkane
42	18.379	0.25	Heptadecanoic acid (Margaric acid)	C17H34O2	270	Saturated fatty acid
43	18.702	1.78	9,12-Octadecadienoic acid (Z,Z)-, methyl ester (Methyl linoleate)	C19H34O2	294	Fatty acid ester
44	18.764	0.97	9-Octadecenoic acid, methyl ester, (E)-	C19H36O2	296	Fatty acid ester
45	18.869	0.05	Neophytadiene	C20H38	278	Terpenoid
46	18.999	0.44	Octadecanoic acid, methyl ester (Methyl stearate)	C19H38O2	298	Fatty acid ester
47	19.066	8.9	10(E),12(Z)-Conjugated linoleic acid	C18H32O2	280	Fatty acid
48	19.118	5.36	Oleic Acid	C18H34O2	282	Fatty acid
49	19.324	4.29	Octadecanoic acid (stearic acid)	C18H36O2	284	Fatty acid ester
50	19.508	0.15	9-Octadecenamide	C18H35NO	281	Fatty amide
51	19.875	0.61	10(E),12(Z)-Conjugated linoleic acid	C18H35NO	281	Fatty amide
52	20.202	0.13	Undec-10-ynoic acid, tetradecyl ester	C25H46O2	378	Fatty acid ester
53	21.076	0.57	Eicosanoic acid (Arachidic acid)	C20H40O2	312	Saturated fatty acid
54	21.925	0.09	1H-1,3-Benzimidazol-7-amine, N-[(4-methoxyphenyl)methyl	C16H17N3O	267	Heterocyclic compound
55	22.291	1.19	Hexadecanoic acid, 2-hydroxy- 1-(hydroxymethyl)ethyl ester (Glycerol beta-palmitate)	C19H38O4	330	Fatty acid ester
56	22.414	0.81	Tyrosine	C9H11NO3	181	Amino acid
57	22.924	0.4	9-Hexacosene	C26H52	364	Hydrocarbon
58	23.005	0.23	1-Decyloxymethyl-3-methyl-1, 3-dihydrobenzoimidazol-2-ylideneamine	C19H31N3O	317	Heterocyclic compounds
59	23.437	0.18	Ethyl 6,9,12,15-octadecatetraenoate	C20H32O2	304	Fatty acid ethyl ester
60	23.661	0.94	9,12-Octadecadienoic acid (Z,Z)-, 2,3-dihydroxypropyl ester	C21H38O4	354	Fatty acid ester
61	23.869	0.17	Octadecanoic acid, 2,3-dihydroxypropyl ester	C21H42O4	358	Fatty acid ester
62	23.91	0.66	Benzoic acid, 2,4-dihydroxy-, 4-(1,1-dimethylpropyl)phenyl	C21H42O4	358	Aromatic ester
63	24.308	0.16	9-octadecenamide (Oleamide)	C18H35NO	281	Fatty amide
64	24.544	0.19	Squalene	C30H50	410	Triterpenoid
65	25.135	0.98	1,54-dibromotetrapentacontane	C54H108Br2	914	Hydrocarbon
66	26.347	0.56	γ-Tocopherol (Vitamin E)	C28H48O2	416	Phenolic compound
67	26.659	0.77	Cholesta-4,6-dien-3-ol, (3.beta.)	C27H44O	384	Cholesterol
68	26.758	1.2	1-Heptacosanol	C27H56O	396	Fatty alcohol
69	26.857	0.4	Stigmast-5-en-3-ol, oleat	C47H82O2	678	Phytosterol
70	27.055	0.49	α -Tocopherol (Vitamin E)	C29H50O2	430	Phenolic compound
71	27.39	0.17	Dodecane, 1,1-dimethoxy-	C14H30O2	230	Acetal
72	27.744	0.27	Tetracosan-10-yl acetate	C26H52O2	396	Fatty acid ester
73	28.224	0.22	Ergost-5-en-3-ol (3.beta.,24R)- (Campesterol)	C28H48O	400	Phytosterol
74	28.509	1.91	Stigmasterol	C29H48O	412	Phytosterol
75	28.662	0.35	Stigmasta-4,22-diene	C29H48	396	Phytosterol
76	28.925	0.99	1-Hexacosanol (Ceryl alcohol)	C26H54O	382	Primary fatty alcohol
77	29.266	10.84	Stigmast-5-en-3-ol, (3beta), (β-sitosterol)	C29H50O	414	Phytosterol
78	29.431	3.82	Stigmastanol	C29H52O	416	Phytosterol
79	29.918	0.7	α-amyrin	C30H50O	426	Triterpenoid
80	30.154	1.44	Lup-20(29)-en-3-one (Lupenone)	C30H48O	424	Triterpenoid
81	30.58	5.09	Lupeol	C30H50O	426	Triterpenoid
82	31.12	1.85	γ Sitostenone	C29H48O	412	Phytosterol
83	32.737	1.45	Cyclopentanone, 2-(5-oxohexyl)	C11H18O2	182	Fatty acid derivative
84	33.938	0.94	Stigmastane-3,6-dione, (5-alpha)	C29H48O2	428	Phytosterol
R. time: Retention time and NR: Not reported

Table 3:

List of phyto-constituents present in green fruits of C. lancifolius analyzed through GC-MS

Peak	R. Time	Area %	Name of the compound	Molecular formula	Molecular weight (g/mol)	Name of compound
1	4.803	0.46	4H-Pyran-4-one, 2, 3-dihydro-3,5-dihydroxy-6-methyl	C6H8O4	144	Phenolic
2	5.913	0.12	Furaneol	C6H8O3	128	Acetate ester
3	6.247	0.37	1,3,5-Triazine-2,4,6-triamine (Melamine)	C3H6N6	126	Heterocyclic compound
4	6.647	0.21	3,7-Dimethylocta-1,6-dien-3-ol (Linalool)	C10H18O	154	Monoterpene alcohol
5	7.173	0.19	Pentanoic acid (Valeric acid)	C18H16O2	144	Alkyl carboxylic acid
6	7.358	2.65	4H-Pyran-4-one, 2,3-dihydro- 3,5-dihydroxy-6-methyl	C6H8O4	144	Phenolic compound
7	7.586	0.03	2-Cyclopenten-1-one, 2-hydroxy- 3-methyl-(Corylon)	C6H8O2	112	Fatty acid derivatives
8	8.604	0.19	5-Hydroxymethylfurfural	C6H6O3	126	Heterocyclic furan derivative
9	8.815	0.07	Butanoic acid, 3-hydroxy-3-methyl	C5H10O3	118	Hydrocarboxylic acid
10	9.113	0.15	Decanoic acid (Capric acid)	C10H20O2	172	Fatty acid
11	9.337	0.05	Butanamide, 2-hydroxy-N,2,3,3-tetramethyl-	C8H17NO2	159	Branched hydroxyamide
12	9.536	0.79	Phenol, 2-methyl-5-(1-methylethyl)	C10H14O	150	Phenolic compound
13	9.667	0.25	Phenol, 2-methyl-5-(1-methylethyl)	C10H14O	150	Phenolic compound
14	9.827	0.13	3-Methylpiperidin-4-ol	C6H13NO	115	Heterocyclic alcohol
15	10.426	0.16	Phenol, 2-methoxy-4-(2-propenyl) (1,3,4-Eugenol)	C10H12O2	164	Phenolic compound
16	10.623	0.48	4-tert-Butylcyclohexyl acetate	C12H22O2	198	Carboxylic ester
17	10.708	26.13	1,2,3-benzenetriol (Pyrogallol)	C6H6O3	126	Phenolic compound
18	11.791	1.78	Silane, dimethyl(but-2-enyloxy) isobutoxy	C10H22OSi	202	Alkoxysilane derivative
19	11.994	0.97	2-Oxovaleric acid, tert-butyldimethylsilyl ester	C11H22O3Si	230	Alpha keto acid
20	12.25	0.24	Anhydro-d-mannosan (Levoglucosan)	C6H10O5	162	Carbohydrate derivative
21	12.683	0.72	10,12-Tricosadiynoic acid	C23H38O2	346	Fatty acid
22	12.868	0.21	2-[Di(tert-butyl) silyl oxy methyl] tetrahydro furane	C13H28O2Si	244	Silyl ether
23	13.019	0.32	Ethyl iso-allocholate	C26H44O5	436	Steroid derivative
24	14.128	0.36	Isocitronellol	C10H20O	156.27	Monoterpenoid
25	14.978	0.24	Caryophyllane, 4,8- beta-epoxy	C15H26O	222	Sesquiterpene
26	15.311	0.66	Tetradecanoic acid (Myristic acid)	C14H28O2	228	Fatty acid
27	15.802	1.4	1-Heptatriacotanol	C37H76O	537	Fatty alcohol
28	16.165	0.83	Neophytadiene	C20H38	278	Terpenoid
29	16.259	0.42	(E)-3-Methyl-5-((1R,4aR,8aR)-5,5,8a-trimethyl -2-methylenedecahydronaphthalen- 1-yl)pent-2-en-1-ol (Copalol)	C20H34O	290	Diterpenoid
30	16.369	1.99	Copalol	C20H34O	290	Diterpenoid
31	16.42	0.29	5-isopropyl-6,6-dimethylhept-3-yne-2,5-diol	C12H22O2	198	Oxygenated monoterpoid
32	16.62	0.8	Phytol (3,7,11,15-Tetramethyl- 2-hexadecen-1-ol)	C20H40O	296	Diterpenoid
33	17.063	0.82	Hexadecanoic acid, methyl ester	C17H34O2	270	Fatty acid ester
34	17.413	11.12	n-Hexadecanoic acid (Palmitic acid)	C16H32O2	256	Fatty acid ester
35	17.817	0.1	Nonadecane	C19H40	268	Alkane
36	18.067	0.18	7-Hexadecenoic acid, methyl ester, (Z)-	C17H32O2	268	Fatty acid ester
37	18.375	0.21	9-octadecenoic acid (z)	C19H36O2	296	Fatty acid ester
38	18.702	0.86	9,12-Octadecadienoic acid (Z, Z), methyl ester (Methyl linoleate)	C19H34O2	294	Fatty acid ester
39	18.764	0.75	9-Octadecenoic acid, methyl ester, (E)	C19H36O2	296	Fatty acid ester
40	18.869	0.14	Phytol (3,7,11,15-Tetramethyl- 2-hexadecen-1-ol)	C20H40O	296	Diterpenoid
41	19.053	2.66	10(E),12(Z)-Conjugated linoleic acid	C18H32O2	280	Fatty acid
42	19.109	2.85	cis-9-Hexadecenal	C16H30O	238	Unsaturated aldehyde
43	19.317	2.46	Octadecanoic acid	C18H36O2	284	Fatty acid ester
44	19.562	0.97	9(E),11(E)-Conjugated linoleic acid (ethyl linoleate)	C18H32O2	280	Fatty acid ethyl ester
45	20.51	0.07	Myristic acid glycidyl ester	C17H32O3	284	Fatty acid ester
46	20.731	0.1	7-Hexadecenal, (Z)-	C16H30O2	238	Unsaturated aldehyde
47	21.077	0.5	9-Octadecenoic acid (Z)-	C19H36O2	296	Fatty acid ester
48	21.345	0.14	Behenic alcohol	C22H46O	326	Fatty alcohol
49	22.192	0.13	Heneicosane	C21H44	296	Hydrocarbon
50	22.29	1.26	Hexadecanoic acid, 2-hydroxy- 1-(hydroxymethyl)ethyl ester	C19H38O4	330	Fatty acid ester
51	22.578	0.22	Docosyl pentafluoro propionate	C25H45F5O2	472	Long-chain fluorinated ester
52	22.72	0.1	1-octanol, 3,7-dimethyl	C10H22O	158	Fatty acid ester
53	22.894	0.03	3-Methylbutylhexa decanoate	C21H42O2	326	Fatty acid ester
54	23.662	0.24	6,9- Octadecadienoic acid, methyl ester	C19H34O2	294	Fatty acid ester
55	23.708	0.16	2-Methylhexacosane	C27H56	380	Hydrocarbon
56	23.867	0.24	Octadecanoic acid, 2,3-dihydroxypropyl ester	C23H42O4	358	Fatty acid ester
57	24.31	0.27	13-Docosenamide, (Z) (Erucamide)	C22H43NO	337	Fatty acid amide
58	25.143	1.95	1-Heptacosanol	C27H56O	396	Fatty alcohol
59	25.899	0.29	Heptacosyl heptafluorobutyrate	C31H55F7O2	592	Hydrocarbon
60	26.344	0.31	γ -Tocopherol	C28H48O2	416	Phenolic compound
61	26.66	0.3	Cholesta-4,6-dien-3-ol, (3-beta)	C27H44O	384	Cholesterol
62	26.757	4.43	1-Heptacosanol	C27H56O	396	Fatty alcohol
63	27.049	0.22	α-Tocopherol	C29H50O2	430	Phenolic compound
64	28.501	1.04	Stigmasta-5,23-dien-3-ol, (3-beta), (Stigmasterol)	C29H48O	412	Phytosterol
65	28.924	1.14	1-Hexacosanol (Ceryl alcohol)	C26H54O	382	Primary fatty alcohol
66	29.259	9.57	β - Sitosterol	C29H50O	414	Phytosterol
67	29.432	2.1	Stigmastanol	C29H52O	416	Phytosterol
68	29.919	0.37	β-Amyrin	C30H50O	426	Triterpenoid
69	30.158	1.31	Lup-20(29)-en-3-one (Lupenone)	C30H48O	424	Triterpenoid
70	30.579	5.57	Lupeol	C30H50O	426	Triterpenoid
71	31.116	1.15	Gamma-sitostenone	C29H48O	412	Phytosterol
R. time: Retention time and NR: Not reported

Table 4:

Phytochemical composition in methanolic extract of dry (Dfr) and green fruits (Gfr) of C. lancifolius

Phytochemical group	Total % area in Dfr	Total % area in Gfr
Fatty acid derivatives	38.09	39.43
Terpenes, terpenoids and derivatives	19.04	16.9
Phenolic compounds and derivatives	9.52	11.26
Phytosterols	9.52	5.63
Hydrocarbons	5.95	8.45
Heterocyclic compounds	5.95	4.22
Amino acid	1.19	-
Miscellaneous compounds	10.71	14.08

The unique compounds which were observed in green fruits as not observed in dry fruits of C. lancifolius are β-Amyrin, Heptacosyl heptafluorobutyrate, Erucamide, 2-Methylhexacosane, Docosyl pentafluoro propionate, 1-octanol, 3,7-dimethyl, 3-Methylbutylhexadecanoate, 6,9- Octadecadienoic acid, methyl ester, 2-Methylhexacosane, Octadecanoic acid, 2,3-dihydroxypropyl ester, 9-Octadecenoic acid (Z)-, Behenic alcohol, Heneicosane, 9(E),11(E)-Conjugated linoleic acid, Myristic acid glycidyl ester, 7-Hexadecenal, (Z)-, cis-9-Hexadecenal, 7-Hexadecenoic acid, methyl ester, (Z)-, Nonadecane, Copalol, 5-isopropyl-6,6-dimethylhept-3-yne-2,5-diol, 1-Heptatriacotanol, Ethyl iso-allocholate, 2-[Di(tert-butyl) silyl oxy methyl] tetrahydro furan, Silane, dimethyl(but-2-enyloxy) isobutoxy, 2-Oxovaleric acid, tert-butyldimethylsilyl ester, Levoglucosan, 3-Methylpiperidin-4-ol, Butanamide, 2-hydroxy-N,2,3,3-tetramethyl-, Butanoic acid, 3-hydroxy-3-methyl, 5-Hydroxymethylfurfural, Corylon, Pentanoic acid, Furaneol, Melamine and Linalool. Whereas the 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, Phytol, Phenol, 2-methyl-5-(1-methylethyl), Copalol, 1-Heptacosanol and 9-Octadecenoic acid (Z)- gave double peaks (Table 3).

DISCUSSION

Due to the ecological and therapeutic importance, research on secondary metabolites has become a key focus in fields such as organic chemistry, pharmacology, molecular biology, and bioinformatics¹⁶. GC-MS is a method that integrates the separation of phytochemicals using gas chromatography with their detection via mass spectrometry and is mostly utilized for identifying and characterizing volatile and semi-volatile components in intricate plant matrices. For phytochemical investigations, the methanolic extracts are mostly used because methanol is an effective solvent for dissolving a variety of polar and moderately non-polar compounds³. Hence, in the present investigation, methanol was utilized for the extraction of maximum phyto-constituents.

The constituents of the methanolic extract of C. lancifolius were classified into major phytochemical groups based on their structural classes as determined by GC-MS analysis (Table 4). Many of these compounds have recognized pharmaceutical significance, with previous studies reporting diverse biological activities, like antioxidant, antidiabetic, anticancer, analgesic, antimicrobial, and anti-inflammatory etc. (Table 2 and 3). Fatty acid derivatives constituted the predominant chemical class, accounting for 38.09% and 39.43% of the total composition in Dfr and Gfr, respectively (Table 4). This predominance primarily resulted from the high abundance of compounds such as palmitic acid, myristic acid, 10(E),12(Z)-conjugated linoleic acid, oleic acid, stearic acid, capric acid, lauric acid, as well as 9-octadecenoic acid methyl ester (E) in Dfr (Table 2) and hexadecanoic acid, 1-heptacosanol, stearic acid, ceryl alcohol, 9-octadecenoic acid methyl ester (E), and cis-9-hexadecenal in Gfr (Table 3).

Fatty acids represent one of the most fundamental classes of biomolecules, exhibiting diverse biological activities with notable therapeutic relevance. Terpenoids, the second most abundant class of biomolecules in the present study (Table 4), are multifunctional secondary metabolites structurally derived from isoprene (C₅) units in plants. C. lancifolius fruits were also found to possess a considerable number of phenolic compounds which have a benzene ring with one or more hydroxyl groups and can occur in a variety of structures such as phenylpropanoids, flavonoids, tannins, melanins, lignins, etc. On the other hand, phytosterols are natural triterpenoids in plants having a tetracyclic structure with functional groups usually at C-3,4,7,12,17 positions. Hydrocarbons are organic compounds made up of carbon and hydrogen atoms and classified as aliphatic/alicyclic or aromatic compounds. Remarkably, all of these secondary metabolites have shown to possess various biological activities for example, antioxidant, antimicrobial, anti-inflammatory, anticancer, antiangiogenic, antidiabetic, hypolipidemic, neuroprotective, immunomodulatory, hepatoprotective etc.^1,17,18. Thus, the rich phytochemical profile of C. lancifolius fruits could motivate pharmacologists for novel drug discovery.

Bonnet et al.¹⁹ have shown that green (immature) fruits of Musa acuminata contained more polyphenol content. Similar results were also observed in this study, where green fruits of C. lancifolius showed more phenolic compounds (11.26%) than dry mature fruits (9.52%). Likewise, higher terpenoid and phytosterol contents were observed in mature fruits of C. lancifolius (28.56%) than in its green fruits (22.53%), as shown in Table 4. These results are similar to a study by Simchuer and Srihanam²⁰where higher triterpenoids and sterol contents were observed in ripe fruits of Ampelocissus martini than in immature fruits. Moreover, high concentrations of β-sitosterol were also found in ripe fruits, comparable to the findings of the present study, where 10.84% β-sitosterol is found to be present in dry mature fruits and 9.57% is found in green fruits.

The compounds detected in the present study have also been reported in other plant parts of C. lancifolius. For example, Al-Shatti et al.²¹ reported 4.90% pyrogallol, 5.61% heneicosane and 5.61% dianhydromannitol from leaves of C. lancifolius. Moni et al.²² also reported various bioactive compounds from hot methanolic extract of the leaves of C. lancifolius through GC-MS, some of which are similar to the present study, such as phytol, hexadecanoic acid, campesterol and oleic acid. The essential oil obtained from leaves of C. lancifolius revealed presence of 85 compounds through GC-MS among which the common compounds in both leaves and fruits are Hexadecanoic acid, Henicosane, Hexacosane, Squalene, γ-Sitosterol, Lupenone, Lupeol, Eicosanoic acid, Neophytadiene, Tetradecanoic acid and Dodecanoic acid with relative abundance % of 2.97, 0.29, 3.06, 1.09, 0.07, 0.61, 3.29, 0.08, 0.26, 0.48, 0.45, respectively as reported by Salim et al.¹⁴.

Interestingly, several phytochemicals detected in the current study have been previously reported in some other plant species of the Combretaceae family, though their relative abundance was variable for example, 4.17% 1-Heptacosanol, 0.5% Hexadecanal, 10.48% γ-Sitosterol were reported in the root of Terminalia travancorensis²³. Recently, Sarvendra et al.²⁴reported various similar phyto-compounds such as 2.66% Phytol, 5.88% Palmitic acid, 2.61% Neophytadiene, 0.12% Heptadecanoic acid, 2.62% Linoleic acid, 0.44% Behenic acid, 1.17% Heptacosane, and 2.31% Squalene from leaves of Combretum indicum. The difference in concentration of these phyto-constituents might be the result of variations at the generic level, edaphic, climatic, and geographical factors, as well as conditions of plant collection and extraction.

Besides, some of these compounds are allelopathic in nature, for example, hexacosane and hexadecanoic acid ethyl ester, which are also found in the leaves of an invasive alien plant species, Lantana camara⁴. This needs to be investigated in detail as C. lancifolius is also an exotic species for India, and questions related to its suitability for plantation in various Indian states are being raised. Therefore, the results of this study may stimulate the assessment of the dual biological roles, i.e., ecological impact and therapeutic potential of C. lancifolius.

CONCLUSION

GC-MS analysis of the methanolic extract of Conocarpus lancifolius fruits unveiled a wide spectrum of bioactive phytochemicals, which have promising therapeutic potential. The major metabolites, particularly lupeol, β-sitosterol acetate, stigmastanol, pyrogallol, linoleic acid, and phytol, dominate the profile and also possess antioxidant, antimicrobial, anti-inflammatory, and lipid-lowering activities. This is the first GC-MS-based comparative study of mature and green (immature) fruits of C. lancifolius and lays the foundation for further bioassay-guided isolation, toxicity assessment, and pharmacological evaluation to find novel drugs.

SIGNIFICANCE STATEMENT

This study provides the first comprehensive evaluation of phyto-constituents from the methanolic extracts of both mature and immature fruits of Conocarpus lancifolius through the GC-MS technique. These phytochemicals are reported to possess a wide range of pharmacological activities. Hence, the present investigation will pave the way for a detailed evaluation of the therapeutic potential of its fruits. Moreover, this study will encourage the researchers to further investigate the allelopathic nature of these compounds, since C. lancifolius is an exotic plant species for India and may become an Invasive Alien Plant Species in the future.

ACKNOWLEDGMENTS

Authors are thankful to the Advanced Instrumentation Research Facility (AIRF), Jawahar Lal Nehru University, New Delhi, India, for providing the GC-MS facility. First author is also thankful to CSIR-UGC, New Delhi, for providing financial assistance in the form of Junior Research Fellowship.

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