Chemical and Biological Investigation of Ceiba chodatii Hassl. Flowers

Abstract

Ceiba (syn. Chorisia) trees have attracted multifaceted attention not only due to their ornamental and economic value but also for their remarkable metabolic diversity and therapeutic properties. In view of that, this work explores the chemical composition of Ceiba chodatii Hassl. and its biological potential. Overall, GC–MS-based analysis of the lipoidal constituents of C. chodatii flowers revealed the presence of diverse classes of metabolites that were dominated by long-chain aliphatic esters (77.016%), ketones (6.396%), aliphatic hydrocarbons (5.757%), fatty alcohols (3.718%), aromatic acid esters (2.794%), alkylamides (1.58%), aldehydes (1.035%), aromatic hydrocarbons (0.31%), and ethers (0.29%). In addition, repeated chromatographic fractionation of different fractions of the total alcoholic extract of the flowers afforded 13 metabolites of varied structural types, including fatty esters and alcohols, phytosterols, monoglycerides, furanoids, and flavonoid glycosides. Structures of the obtained compounds were determined by different spectroscopic techniques, such as 1H- and 13C-NMR, APT, DEPT, and EI–MS analyses. Noteworthily, a wide range of the metabolites identified herein using different analytical approaches were described for the first time in the plant species under study or in those belonging to the genus Ceiba. Finally, the total extract and different fractions of C. chodatii flowers as well as the isolated flavonoids showed weak anti-infective potential against a group of human pathogens at concentration ranges up to 200 and 20 µg/mL, respectively. In contrast, the total extract and different fractions of the flowers exerted mild to moderate anti-proliferative activities against MDA-MB-468 cells, with IC₅₀ in the range of 21.69–47.60 μg/mL.

1. Introduction

Plants are prolific sources of a plethora of natural products with privileged chemical diversity. Such bioactive phytocompounds have gained noteworthy attention thanks to their various therapeutic effects and potential biomedical applications, offering a treasure trove for the discovery of new drug leads [1]. With the increasing focus on natural remedies, unraveling the chemical complexity of plants has been made possible thanks to the great advances in analytical technologies [2]. Of these, gas chromatography (GC) has emerged as one of the most powerful, versatile, and commonly used tools for analyzing chemical mixtures [3]. This technique allows the high-throughput profiling of primary and secondary metabolites from plants comprising their volatile, lipophilic, as well as derivatized hydrophilic components [3]. Over the years, the coupling of GC with mass spectrometry (GC–MS) has also become a valuable investigative tool that enabled the annotation of several phytomolecules in mixtures, such as plant extracts or fractions, owing to the existence of several spectral databases and the reproducible fragmentation profiles obtained [3,4]. Such analytical integration has extended the praiseworthy role of GC in phytomedicine, food science, agriculture, and other plant-based research arenas [5].

Human pathogens and parasites hold the potential to cause a wide spectrum of health problems ranging from minor infections to chronic and life-threatening illnesses [6]. Despite the significant progress in human medicine, infectious diseases are still among the major threats to public health with increasing mortality rates, especially in tropical and low-income countries due to the relative lack of medicines and the emergence of multidrug resistance [7]. In this respect, many epidemiological studies have indicated the development of widespread resistance to different antimicrobials, including not only antibiotics but also antifungal and anti-parasitic medications [8]. Therefore, the search for naturally based, potent, and cost-effective anti-infective agents has stimulated substantial research throughout the past decades [7,9]. In this vein, medicinal plants have been proven to enjoy such desired advantages, and thus the screening of the anti-infective efficacy of plant-derived natural products has been widely addressed [7,10].

Ceiba Mill. (syn. Chorisia Kunth), commonly known as silk floss or bottle trees, is a genus of deciduous trees native to the tropics and subtropics. It belongs to the plant family Bombacaceae, which has been lately merged into the Malvaceae, and includes about twenty species [11]. Aside from their ornamental and economic importance, plants of this genus are effective natural remedies in the management of diarrhea, peptic ulcers, diabetes, headache, rheumatism, fever, and parasitic infections [12]. Earlier phytochemical studies have revealed the ability of Ceiba species to accumulate a myriad of polyphenolic metabolites like flavonoids, anthocyanins, phenolic acids, lignans, and coumarins, along with a number of furanoids, quinones, megastigmanes, sterols, terpenoids, fatty acids, and organic acids [12]. Moreover, different extracts and purified metabolites from some Ceiba species have been reported to possess hypoglycaemic, cytotoxic, antioxidant, hepatoprotective, antipyretic, antimicrobial, and anti-inflammatory activities [12,13].

Inspired by the interesting metabolic diversity and medicinal potential of Ceiba plants and in continuation of our search for potent anti-infective agents from natural sources, the present work aims at (i) applying gas chromatography–mass spectrometry (GC–MS)-based metabolomics to explore the overlooked lipophilic metabolites of Ceiba chodatii Hassl. (syn. Chorisia chodatii (Hassl.) Ravenna) flowers, (ii) studying the chemical composition of various extracts of C. chodatii flowers through chromatographic isolation and structure elucidation of their phytoconstituents, (iii) evaluating the antibacterial, antifungal, antileishmanial, and antimalarial potential of flowers, and (iv) testing for the anti-proliferative potential of the flowers against triple-negative breast cancer (TNBC).

2. Materials and Methods

2.1. General Experimental Procedures

Different solvents used in this work were purchased from El-Nasr Company for Pharmaceuticals and Chemicals, Cairo, Egypt. NMR spectroscopic analyses were carried out on Bruker Avance HD III 400 and 600 MHz (Bruker, Uster, Switzerland) spectrometers using deuterated solvents, e.g., DMSO-d₆, CD₃OD, and CDCl₃ (Sigma-Aldrich, Darmstadt, Germany). Electron impact (EI) mass analyses were obtained on a Thermo Scientific mass spectrometer (Thermo Fisher Scientific, Austin, TX, USA). Column chromatography (CC) was performed on silica gel 60 (60–120 mesh; E. Merck, Darmstadt, Germany) and sephadex LH–20 (0.25 mm; Sigma-Aldrich, Darmstadt, Germany), whereas silica gel GF254 for thin layer chromatography (TLC; El-Nasr Company for Pharmaceuticals and Chemicals, Cairo, Egypt) was employed for vacuum liquid chromatography (VLC). HPLC purifications were performed on a semi-preparative ODS column (Inertsil ODS-3; GL Science, Tokyo, Japan; Φ = 10 mm and L = 25 cm) at 2.0 mL/min using HPLC-grade solvents (SDFCL sd Fine-Chem Limited, Mumbai, India) and a refractive index detector (JASCO RI-930, Tokyo, Japan). Pre-coated silica gel 60 GF₂₅₄ plates (E. Merck, Darmstadt, Germany; 20 × 20 cm, 0.25 mm-thick) were used for TLC work and visualized by spraying with a solution of 10% H₂SO₄ in ethanol and heating at 110 °C.

2.2. Plant Material

The fresh flowers were collected from the campus of Minia University, Egypt, and kindly identified by Prof. Ahmed Abdel-Monem (Horticulture Department, Faculty of Agriculture, Minia University, Minia, Egypt). A voucher sample was deposited in the Pharmacognosy Department, Faculty of Pharmacy, Minia University, Egypt under the number (Mn-Ph-Cog-001).

2.3. Extraction and Fractionation

The total ethanol extract of C. chodatii flowers and its fractions (I–IV) were obtained as described in Figure 1 using 70% ethanol, distilled water, ethyl acetate, chloroform, and light petroleum ether (b.p. 60–80 °C).

2.4. GC–MS Analysis

GC–MS analysis was performed on a GC–MS–QP2010SE model (Shimadzu Scientific Instruments, INC, serial no. O215355, Columbia, MD, USA) equipped with a DB-5-MS column (30 m × 0.25 mm; film thickness = 0.25 μm) and a flame ionization detector. Helium was the carrier gas at a flow rate of 1 mL/min. The injector port and detector temperatures were set at 250 °C and the oven temperature was programmed at 50–300 °C at a rate of 5 °C/min. Electron impact mass spectra were obtained at an ionization voltage of 70 eV over a range of 50–500 m/z and an ion source temperature of 200 °C. Relative amounts of the detected compounds were obtained from the total ion chromatogram, and their identification was carried out via the comparison of their mass spectra with a set of databases, encompassing National Institute of Standards and Technology (NIST), Replib, Mainlib, and Wiley library 09 [14,15].

2.5. Isolation of Compounds 1–13

A part of fraction I (70.0 g) was subjected to VLC on silica gel (500 g; 6.5 × 30 cm) for two times (35 g each) using petroleum ether–ethyl acetate (100:0, 90:10, 80:20, 50:50, 20:80, and 0:100) to provide six subfractions (I1−I6). Subfraction I2 (17.2 g) was then chromatographed on a silica gel column using gradient mixtures of petroleum ether–ethyl acetate (100:0, 98:2, 95:5, 90:10, 85:15, 80:20, and 0:100) to yield seven subfractions (I2-F-1−I2-F-7). Of these, I2-F-3 was further purified by silica gel CC using gradient elution with petroleum ether–chloroform (100:0, 99:1, 98:2, 97:3, 94:6, 88:12, 86:14, 84:16, 80:20, 75:25, and 0:100) to give compounds 1 (15.7 mg) and 2 (46.2 mg) as yellow oils from I2-F-3-3 and I2-F-3-8, respectively. Subfraction I2-F-3-9 was similarly re-purified by silica gel CC using petroleum ether–benzene (100:0, 100:1, 100:10, 100:15, and 100:20), providing compounds 3 (38.7 mg) and 4 (25.7 mg) as yellow oils from I2-F-3-9-4 and I2-F-3-9-5, respectively, whereas compound 5 (40.0 mg) was precipitated from subfraction I2-F-5 as white amorphous powder. Subfraction I2-F-6 was also subjected to silica gel CC using gradient mixtures of petroleum ether–chloroform (40:60, 30:70, 20:80, 10:90, 5:95, and 0:100) to obtain I2-F-6-1–I2-F-6-6. Compound 6 (39.6 mg) was obtained as yellow oil after repeated purification of I2-F-6-5 on silica gel using mixtures of petroleum ether–ethyl acetate (85:15, 82:18, 80:20, and 0:100), then on Sephadex using chloroform–methanol (50:50) as the mobile phase.

On the other hand, fraction II (12.0 g) was fractionated by silica gel VLC (85 g; 6.5 × 30 cm), applying gradient elution with petroleum ether–ethyl acetate (80:20, 60:40, 40:60, 20:80, and 0:100) and finally washed with methanol to obtain six subfractions (II1−II6). Among them, subfraction II4 (1.1 g) was further subjected to silica gel CC using chloroform–methanol (97:3, 95:5, and 90:10) to afford compound 7 (40.0 mg) from II4-F-3 as white amorphous powder.

Likewise, VLC fractionation of 40.0 g of fraction III on silica gel (500 g; 6.5 × 30 cm) using petroleum ether–ethyl acetate (70:30, 50:50, 20:80, and 0:100), then ethyl acetate–methanol (90:10 and 0:100) afforded six subfractions (III1−III6). Of these, subfraction III2 (3.2 g) was subjected to silica gel CC using petroleum ether–ethyl acetate (80:20, 70:30, 65:35, 60:40, and 0:100) to yield five subfractions (III2-F-1−III2-F-5), of which III2-F-1 provided compound 8 (15.2 mg) as yellow oil. Similarly, silica gel CC fractionation of III5 (10.0 g) using gradient mixtures of ethyl acetate–methanol (100:0, 95:5, 90:10, 70:30, and 0:100) provided five subfractions (III5-F-1−III5-F-5). Among them, subfraction III5-F-1 was re-chromatographed on silica gel using gradient elution with petroleum ether–ethyl acetate (20:80, 10:90, and 0:100), then ethyl acetate–methanol (98:2 and 0:100) to obtain III5-F-1-1–III5-F-1-5. HPLC purification of III5-F-1-2 using water–acetonitrile afforded compounds 9 (5.0 mg) and 10 (8.0 mg) as brown residues, whereas compound 11 (20.3 mg) was obtained as a yellow powder from III5-F-1-3 by precipitation. Moreover, III5-F-1-4 was further purified by silica gel CC using ethyl acetate–methanol gradient mixtures (100:0, 99:1, and 98:2) to yield compound 12 (12.0 mg) from III5-F-1-4-1 as a yellow powder. Finally, compound 13 (850.0 mg) was similarly obtained as yellow powder by silica gel CC purification of III5-F-3 using ethyl acetate–methanol gradient elution (100:0, 95:5, 93:7, 90:10, 80:20, and 0:100), namely from subfraction III5-F-3-3 (Figure 1).

2.6. Antimicrobial Activity

The total extract, different fractions, and isolated flavonoids of C. chodatii flowers were tested for their antimicrobial potential against a panel of standard strains of bacteria (e.g., methicillin-resistant Staphylococcus aureus ATCC 43300 (MRSa), Klebsiella pneumoniae ATCC 10031, Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, and vancomycin-resistant Enterococcus faecalis ATCC 49533 (VR)) and fungi (e.g., Aspergillus fumigatus ATCC 90906, Cryptococcus neoformans ATCC 90113, Candida albicans ATCC 90028). Susceptibility testing was carried out using a modified version of the CLSI methods [16,17]. The tested samples were serially diluted in 20% DMSO/saline (8–200 µg/mL for the total extract and fractions (I–IV) and 0.8–20 µg/mL for the purified compounds), then transferred in duplicate to 96-well flat-bottom microplates. Microbial inocula were prepared by adjusting the OD630 of microbe suspensions in incubation broth to provide the final inocula. Amphotericin B and ciprofloxacin (ICN Biomedicals, Aurora, OH, USA) were included as positive antifungal and antibacterial controls, respectively. The optical densities were measured at 630 nm using an EL-340 Biokinetics reader (Bio-Tek Instruments, Winooski, VT, USA) or 544 (excitation)/590 (emission) nm (for A. fumigatus) using a Polarstar Galaxy plate reader (BMG LabTechnologies, Ortenberg, Germany) before and after incubation. IC₅₀ values were finally obtained by plotting the percent growth versus test concentration [16].

2.7. Antileishmanial Activity

The in vitro inhibitory potential of the total extract, different fractions, and purified flavonoids of C. chodatii against Leishmania donovani was tested following the protocol of Mahmoud et al. [16]. In brief, the promastigotes were initially grown at 26 °C in RPMI 1640 medium enriched with 10% fetal calf serum (Gibco Chem. Co., Carlsbad, CA, USA). The three-day-old culture was then diluted to 5 × 10⁵ promastigotes/mL. Sample dilutions were directly made in cell suspension in 96-well plates that were further incubated at 26 °C for 48 h. A final concentration range of 0.8–20 µg/mL was used for the total extract and fractions (I–IV), while the purified compounds were tested at 0.4–10 µg/mL. The growth of promastigotes was then evaluated by the Alamar BlueTM assay. Standard fluorescence was measured using a FLUOstar Galaxy plate reader (BMG LabTechnologies, Ortenberg, Germany) at excitation and emission wavelengths of 544 and 590 nm, respectively. Pentamidine (ICN Biomedicals, Aurora, OH, USA) served as a standard antileishmanial agent. Dose-response curves were prepared to calculate IC₅₀ values.

2.8. Antimalarial Activity

The antimalarial activity was evaluated in vitro against chloroquine-resistant (W2, Indochina) and chloroquine-sensitive (D6, Sierra Leone) strains of Plasmodium falciparum by measuring plasmodial lactate dehydrogenase (PLDH) activity as described by Makler and Hinrichs and Mahmoud et al. [16,18]. Different fractions and the isolated flavonoids of C. chodatii flowers were initially prepared in DMSO (2 mg/mL). A 200 µL of the culture suspension of P. falciparum [2% parasitemia and 2% hematocrit in RPMI 1640 medium containing 10% human serum and amikacin (60 µg/mL)] was added to 96-well plates containing 10 µL of the serially diluted plant samples. The plates were then flushed with a mixture of 5% O₂, 5% CO₂, and 90% N2, and then incubated at 37 °C for 72 h. PLDH activity was assessed by a MalstatTM reagent (Flow Inc., Portland, ON, USA). Briefly, a volume of 20 µL of the incubated mixtures was mixed with 100 µL of the reagent and re-incubated for 30 min. Next, 20 µL of a 1:1 mixture of NBT/PES (Sigma, St. Louis, MO, USA) was added, followed by incubation for further 1 h in the dark. A volume of 100 µL of 5% acetic acid was finally added to stop the reaction, and the plates were read at 650 nm using an EL-340 Biokinetics reader. Chloroquine and 0.25% DMSO were employed as positive and vehicle controls, respectively. IC₅₀ values were obtained by plotting the percent growth versus test concentration.

2.9. Anti-Proliferative Activity

The anti-proliferative potential was examined against MDA-MB-468 TNBC (African American) cell line obtained from the American Type Culture Collection, Manassas, VA, USA, using the MTT assay as reported by Mendonca et al. [19]. Plant extracts (dissolved in DMSO) were added to 96-well plates at 50, 25, and 12.5 μg/mL. Cells were then incubated for 72 h at 37 °C, and a volume of 100 μL of the MTT solution was added, followed by incubation for further 1.5 h. The fluorescence signal was measured (at 540/580 nm) by a microplate reader. Control wells were also treated with DMSO at the same concentration used for the extracts (< 1%). The blank wells involved only the culture media without cells. The cytotoxic activities were determined as: % inhibition = [1 − (A_sample − A_blank)/(A_control − A_blank)] × 100. Finally, IC₅₀ values (in μg/mL) ± standard deviation (S.D.) were obtained using Graph Pad Prism (version 6.07) and compared with doxorubicin as a positive control.

3. Results and Discussion

3.1. GC–MS Profiling of C. chodatii Flowers

Previous chemical investigations of C. chodatii led to the characterization of a limited number of flavonoids, phenolic acids and esters, coumarins, furanoids, and megastigmane glycosides, along with a variety of volatile components [12], while the lipophilic metabolites accumulated by this plant species have not been previously deliberated. Therefore, different constituents of the petroleum ether fraction of C. chodatii flowers were profiled herein using the GC–MS technique (Supplementary Material Figure S1). The obtained data unveiled the presence of 82 components of varied structural types, of which 74 compounds—representing 98.896%—were identified, while only eight compounds (1.104%) were not identified (

Table 1. Compounds identified in the petroleum ether fraction of C. chodatii flowers.

Peak No.	Compound	Molecular Formula	Rt a (min)	RRt b	Peak Area (%)	Main Fragment Ions (m/z) c
1	Methyl 9-oxononanoate *	C10H18O3	14.060	0.558	0.052	186, 171, 158, 155, 143, 136, 115, 111, 97, 87, 74, 67, 59, 55, 41
2	Methyl 4-hydroxy benzoate *	C8H8O3	14.765	0.586	0.599	152, 121, 107, 93, 65, 50, 39
3	1-Methyl-2-heptenyl 2,6-difluorobenzoate *	C15H18F2O2	15.540	0.617	0.305	268, 181, 157, 141, 127, 111, 81, 68, 55
4	Methyl vanillate *	C9H10O4	16.055	0.637	0.194	182, 167, 151, 136, 123, 108, 93, 79, 65, 51
5	Methyl dodecanoate (methyl laurate) *	C13H26O2	16.225	0.644	0.115	214, 199, 183, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
6	Ethyl 2-methylallyl fumarate *	C10H14O4	17.365	0.690	0.505	198, 153, 127, 117, 99, 82, 69, 55, 39
7	Methyl 10-hydroxy-11-dodecenoate *	C13H24O3	18.205	0.723	0.039	228, 210, 196, 181, 172, 143, 129, 111, 98, 87, 74, 69, 55, 41
8	Methyl tetradecanoate (methyl myristate) *	C15H30O2	20.775	0.825	0.628	242, 211, 199, 185, 171, 157, 143, 129, 101, 87, 74, 55, 41
9	3,7,11,15-Tetramethylhexadecyl acetate *	C22H44O2	20.985	0.833	0.124	340, 280, 252, 210, 196, 154, 140, 126, 111, 97, 83, 69, 57, 43
10	Methyl 4-hydroxycinnamate *	C10H10O3	21.540	0.855	0.431	178, 160, 147, 133, 119, 107, 91, 65, 50
11	Methyl 4-hydroxy-3,5-dimethoxy benzoate *	C10H12O5	21.720	0.863	0.071	212, 197, 181, 165, 153, 137, 123, 108, 93, 79, 67, 50, 39
12	1,1-Dimethoxy octadecane *	C20H42O2	22.305	0.886	0.150	314, 283, 250, 229, 197, 180, 165, 138, 123, 111, 96, 81, 75, 71, 55
13	Methyl (5Z)-dodec-5-enoate *	C13H24O2	22.550	0.896	0.048	212, 194, 180, 165, 138, 123, 110, 96, 81, 74, 67, 55, 41
14	Methyl pentadecanoate	C16H32O2	22.895	0.909	0.341	256, 241, 225, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
15	Unidentified	–	23.115	0.918	0.036	278, 263, 249, 236, 222, 204, 179, 167, 152, 138, 123, 109, 95, 82, 68, 57, 43
16	6,10,14-Trimethyl-2-pentadecanone (Hexahydrofarnesyl acetone)	C18H36O	23.270	0.924	1.475	268, 250, 235, 225, 210, 194, 179, 165, 151, 137, 124, 109, 95, 85, 71, 58, 43
17	Diisobutyl phthalate *	C16H22O4	23.615	0.938	0.226	278, 263, 223, 205, 189, 167, 149, 132, 121, 104, 93, 76, 57, 41
18	2-Heptadecanone	C17H34O	24.505	0.973	4.200	254, 239, 225, 211, 196, 180, 166, 152, 138, 127, 110, 96, 85, 71, 58, 43
19	n-Heptadecyl trifluoroacetate *	C19H35F3O2	24.705	0.981	0.417	352, 341, 238, 210, 196, 182, 168, 154, 140, 125, 111, 97, 83, 69, 57, 43
20	Methyl hexadecanoate (methyl palmitate)	C17H34O2	25.165	1.000	28.185	270, 255, 239, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
21	Dibutyl phthalate **	C16H22O4	25.565	1.015	0.193	278, 223, 205, 149, 135, 121, 104, 93, 76, 57
22	Methyl (9Z, 12Z)-hexadeca-9,12-dienoate *	C18H32O2	26.235	1.042	0.216	280, 251, 206, 192, 177, 164, 150, 135, 121, 109, 95, 81, 67, 55, 41
23	Methyl (9Z)-heptadec-9-enoate *	C18H34O2	26.365	1.047	0.180	282, 267, 251, 232, 221, 208, 194, 166, 152, 137, 123, 110, 97, 83, 69, 55, 41
24	Methyl 2-hexylcyclopro-paneoctanoate *	C18H34O2	26.570	1.055	0.185	282, 250, 232, 221, 208, 194, 180, 166, 152, 138, 123, 110, 96, 83, 74, 69, 55, 41
25	Methyl heptadecanoate (methyl margarate)	C18H36O2	26.885	1.068	0.986	284, 269, 253, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
26	Methyl 8-octadecynoate *	C19H34O2	27.070	1.075	0.076	294, 263, 245, 220, 196, 180, 164, 150, 135, 122, 108, 95, 81, 67, 55, 41
27	Methyl (12E,15E)-octadeca-12,15-dienoate *	C19H34O2	28.380	1.127	10.676	294, 279, 263, 235, 220, 192, 178, 164, 150, 136, 123, 109, 96, 81, 67, 55, 41
28	Methyl (9Z,12Z,15Z)-octadeca-9,12, 15-trienoate (methyl linolenate)	C19H32O2	28.495	1.132	9.211	292, 277, 264, 250, 236, 222, 207, 191, 180, 163, 149, 135, 121, 108, 95, 79, 67, 55, 41
29	Phytol	C20H40O	28.600	1.136	3.668	296, 278, 263, 249, 210, 196, 179, 165, 140, 123, 111, 95, 81, 71, 57, 43
30	Methyl octadecanoate (methyl stearate)	C19H38O2	28.880	1.147	9.254	298, 283, 267, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
31	Methyl 9-octadecynoate *	C19H34O2	28.975	1.151	0.104	294, 263, 245, 210, 196, 178, 164, 152, 136, 122, 109, 95, 81, 67, 55, 41
32	Phytol acetate *	C22H42O2	29.170	1.159	0.028	338, 278, 263, 236, 223, 208, 193, 179, 151, 137, 123, 109, 95, 81, 68, 57, 43
33	Ethyl (9Z,12Z)-octadeca-9,12-dienoate (ethyl linoleate) *	C20H36O2	29.360	1.166	0.078	308, 293, 279, 263, 220, 205, 191, 178, 164, 150, 136, 123, 109, 95, 81, 67, 55, 41
34	Ethyl (9Z,12Z,15Z)-octadeca-9,12, 15-trienoate (ethyl linolenate) *	C20H34O2	29.465	1.170	0.037	306, 291, 277, 264, 250, 237, 222, 207, 191, 173, 163, 149, 135, 121, 108, 95, 79, 67, 55, 41
35	Unidentified	–	29.695	1.180	0.097	281, 266, 251, 236, 221, 213, 196, 183, 170, 154, 140, 126, 112, 98, 86, 72, 59, 41
36	Hexadecanamide (palmitamide) *	C16H33NO	30.005	1.192	0.315	255, 226, 212, 198, 184, 170, 156, 142, 128, 114, 100, 86, 72, 59, 43
37	n-Docosane	C22H46	30.065	1.194	0.134	310, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
38	Methyl (10Z)-nonadec-10-enoate *	C20H38O2	30.245	1.201	0.295	310, 295, 278, 260, 249, 236, 221, 208, 194, 180, 166, 152, 139, 125, 111, 97, 83, 74, 69, 55, 41
39	Neophytadiene *	C20H38	23.115	0.918	0.036	278, 249, 236, 222, 208, 174, 179, 165, 152, 138, 123, 109, 95, 81, 68, 55, 41
40	Methyl nonadecanoate *	C20H40O2	30.540	1.213	0.202	312, 281, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
41	(Z)-7-Tetradecenal *	C14H26O	32.545	1.293	0.777	210, 192, 149, 135, 121, 111, 97, 83, 67, 55, 41
42	Tributyl acetylcitrate *	C20H34O8	30.930	1.229	0.088	402, 329, 273, 259, 213, 185, 157, 147, 129, 111, 57, 43
43	(E)-10,13,13-Trimethyl-11-tetradecenyl acetate *	C19H36O2	31.150	1.237	0.312	296, 281, 239, 222, 166, 151, 123, 109, 95, 83, 71, 57, 43
44	(Z)-9-Hexadecenal *	C16H30O	30.790	1.223	0.095	238, 220, 194, 179, 163, 149, 135, 121, 111, 95, 81, 69, 55, 41
45	Unidentified	–	31.475	1.250	0.075	292, 278, 261, 249, 235, 221, 200, 185, 168, 153, 135, 125, 107, 103, 93, 79, 69, 55
46	S-(tert-Butyl) (9E)-12-hydroxy-9-octadecenethioate *	C22H42O2S	31.615	1.256	1.704	370, 296, 279, 261, 199, 167, 149, 139, 121, 113, 95, 83, 71, 57, 41
47	Unidentified	–	31.675	1.258	0.199	322, 290, 199, 167, 149, 139, 121, 107, 93, 79, 67, 57
48	n-Tricosane **	C23H48	31.775	1.262	1.374	324, 267, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
49	2-Octadecanone *	C18H36O	31.920	1.268	0.519	268, 253, 239, 208, 194, 180, 166, 152, 138, 127, 111, 96, 85, 71, 58, 43
50	Methyl eicosanoate (methyl arachidate)	C21H42O2	32.295	1.283	3.187	326, 295, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
51	(E)-Tridec-2-yn-1-yl hept-2-enoate *	C20H34O2	32.370	1.286	0.647	306, 277, 263, 249, 221, 193, 179, 166, 149, 135, 111, 93, 81, 67, 55, 41
52	(Z)-9-Octadecenal (olealdehyde) *	C18H34O	31.300	1.243	0.069	266, 248, 222, 207, 194, 163, 149, 135, 121, 111, 98, 81, 69, 55, 41
53	Unidentified	–	32.610	1.295	0.221	290, 254, 236, 221, 207, 185, 179, 151, 125, 108, 99, 79, 69, 55
54	(Z)-13-Octadecenal *	C18H34O	32.700	1.299	0.094	266, 248, 227, 183, 167, 149, 135, 121, 110, 97, 83, 69, 55, 41
55	Methyl (9E, 12E)-octadeca-9,12-dienoate (methyl linolelaidate) *	C19H34O2	32.765	1.302	0.062	294, 279, 263, 220, 178, 164, 149, 135, 121, 109, 95, 81, 67, 55, 41
56	(Z)-9-Octadecenamide (oleamide) *	C18H35NO	32.890	1.306	1.265	281, 264, 238, 222, 184, 154, 140, 126, 112, 98, 86, 72, 59, 41
57	Methyl 2-octylcyclopropene-1-octanoate *	C20H36O2	32.945	1.309	0.094	308, 277, 263, 249, 235, 223, 209, 195, 177, 164, 151, 135, 121, 105, 95, 81, 67, 55, 41
58	2,6-Dimethyldecahydronaphthalene *	C12H22	33.095	1.315	0.153	166, 151, 137, 123, 109, 95, 81, 67, 55, 41
59	1(22),7(16)-Diepoxy-tricyclo[20.8.0.0(7,16)] triacontane *	C30H52O2	33.185	1.318	0.140	444, 310, 292, 278, 261, 236, 221, 207, 185, 179, 164, 153, 135, 121, 109, 95, 81, 67, 55, 41
60	But-3-yn-1-yl octadecyl carbonate *	C23H42O3	33.245	1.321	0.052	366, 334, 319, 295, 276, 252, 239, 210, 196, 182, 168, 153, 139, 125, 115, 97, 83, 69, 55, 43
61	n-Tetracosane	C24H50	33.395	1.327	0.388	338, 309, 295, 281, 267, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
62	Methyl heneicosanoate *	C22H44O2	33.880	1.346	0.820	340, 309, 297, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
63	Unidentified	–	33.975	1.350	0.061	264, 187, 155, 122, 109, 93, 79, 67, 55
64	Tridec-2-ynyl 2,6-difluorobenzoate *	C20H26F2O2	34.065	1.353	0.410	336, 277, 250, 235, 184, 166, 141, 107, 93, 79, 67, 55, 41
65	2,3-Dimethyldecahydronaphthalene *	C12H22	34.165	1.357	0.157	166, 151, 137, 123, 109, 95, 81, 67, 55, 41
66	(Z)-12-Pentacosene *	C25H50	34.560	1.373	0.180	350, 322, 308, 294, 278, 252, 236, 209, 195, 181, 167, 153, 139, 125, 111, 97, 83, 69, 57, 43
67	1-Eicosanol (arachidic alcohol) *	C20H42O	34.880	1.386	0.050	298, 252, 236, 196, 181, 167, 153, 139, 125, 111, 97, 83, 69, 55, 43
68	n-Pentacosane	C25H52	35.005	1.391	2.355	352, 323, 309, 295, 281, 267, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
69	2-Nonadecanone	C19H38O	35.165	1.397	0.202	282, 264, 236, 222, 208, 194, 180, 166, 152, 127, 110, 96, 85, 71, 58, 43
70	2-Monopalmitin *	C19H38O4	35.285	1.402	0.751	330, 312, 299, 270, 257, 239, 227, 213, 196, 182, 168, 147, 134, 112, 98, 84, 74, 57, 43
71	Methyl docosanoate (methyl behenate)	C23H46O2	35.520	1.411	4.026	354, 339, 323, 311, 297, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
72	Unidentified	–	36.100	1.434	0.180	240, 225, 208, 193, 171, 155, 150, 134, 121, 107, 97, 83, 72, 67, 55
73	n-Hexacosane	C26H54	36.660	1.456	0.200	366, 337, 323, 309, 295, 281, 267, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
74	Methyl tricosanoate *	C24H48O2	37.260	1.480	0.882	368, 353, 337, 325, 311, 297, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
75	n-Octadecyl trifluoroacetate *	C20H37F3O2	38.140	1.515	0.084	366, 293, 279, 264, 251, 237, 208, 194, 181, 167, 153, 139, 125, 111, 97, 83, 69, 57, 43
76	1-Monolinolein *	C21H38O4	38.600	1.533	0.822	354, 337, 294, 280, 262, 245, 234, 220, 205, 191, 177, 163, 149, 135, 121, 109, 95, 81, 67, 55, 41
77	n-Heptacosane *	C27H56	38.770	1.540	1.090	380, 365, 351, 337, 323, 309, 295, 281, 267, 253, 239, 225, 211, 197, 183, 169, 155, 141, 127, 113, 99, 85, 71, 57, 43
78	Methyl tetracosanoate (methyl lignocerate) *	C25H50O2	39.515	1.570	1.115	382, 351, 339, 325, 311, 297, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
79	Unidentified	–	40.305	1.601	0.235	430, 414, 296, 278, 263, 249, 235, 222, 205, 190, 179, 165, 152, 137, 123, 109, 95, 82, 68, 57, 43
80	Bis(7-methyloctyl) phthalate *	C26H42O4	41.605	1.653	0.365	418, 293, 275, 167, 149, 127, 98, 85, 71, 57, 43
81	Methyl pentacosanoate *	C26H52O2	42.270	1.679	0.179	396, 365, 353, 339, 325, 311, 297, 283, 269, 255, 241, 227, 213, 199, 185, 171, 157, 143, 129, 115, 101, 87, 74, 55, 41
82	n-Eicosyl trifluoroacetate *	C22H41F3O2	44.805	1.780	0.209	394, 379, 365, 278, 227, 213, 199, 182, 167, 153, 139, 125, 111, 97, 83, 69, 57, 43
Percentage of oxygenated compounds					92.829%
Percentage of non-oxygenated compounds (hydrocarbons)					6.067%
Percentage of total identified compounds					98.896%
Percentage of total unidentified compounds					1.104%

^a R_t: retention time. ^b RR_t: relative retention time (relative to the major identified compound methyl palmitate (peak no. 20)). ^c Underlined m/z values indicate base peaks. * Compounds are reported herein for the first time in the genus. ** Compounds are reported herein for the first time in the species.

). The characterized metabolites were largely prevailed by oxygenated principles (64 compounds; 92.829%), whereas only ten non-oxygenated compounds (hydrocarbons) were detected that accounted for 6.067% of the total identified metabolites (Table 1 and Supplementary Material Table S1). Based on their chemical nature, the group of oxygenated metabolites comprised six classes, namely esters (79.81%)—the most predominant group—, long-chain ketones (6.396%), long-chain alcohols (3.718%), alkylamides (1.58%), aldehydes (1.035%), and ethers (0.29%) (Supplementary Material Table S1) with methyl palmitate (28.185%), 2-heptadecanone (4.200%), phytol (3.668%), oleamide (1.265%), 7-tetradecenal (0.777%), and 1,1-dimethoxy octadecane (0.150%) were the major representatives of each category, respectively. In the same regard, the identified esters in the lipid fraction of the flowers were dominated by aliphatic molecules (41 esters), which constituted 77.016% of the total characterized metabolites. This group was mainly represented by a range of long-chain fatty esters of both different chain lengths and degrees of unsaturation, such as methyl palmitate (28.185%), methyl octadeca-12,15-dienoate (10.676%), methyl stearate (9.254%), methyl linolenate (9.211%), methyl behenate (4.026%), and methyl arachidate (3.187%), along with monoacylglycerols (1.573%). Aromatic acid esters were detected as well, but at a remarkably lesser proportion (9 compounds; 2.794%), including a number of benzoic, vanillic, cinnamic, and phthalic acid derivatives. On the other hand, the assortment of non-oxygenated metabolites (6.067%) in C. chodatii flowers consisted of eight long-chain hydrocarbons (C20–C27; 5.757%), of which n-pentacosane (2.355%) was the major constituent, together with two methylated naphthalene derivatives (0.31%) (Table 1 and Supplementary Material Table S1). Noteworthily, this work is the first investigation of the lipoidal metabolites of C. chodatii; therefore, a wide section of the identified metabolites is described herein for the first time either in the plant species under study or in those belonging to the genus Ceiba as indicated in Table 1.

3.2. Identification of Compounds 1–13

Repeated chromatographic fractionation of different fractions of the total extract of C. chodatii flowers afforded 13 compounds of varied structural types (Figure 2). By comparing their spectral data with the literature (Supplementary Data), the obtained metabolites were identified as follows: hexadecan-4-yl hexanoate (1), ethyl hexadecanoate (2) [20], ethyl (9Z)-octadec-9-enoate (ethyl oleate) (3) [21], 14-octacosanol (4), a mixture of β-sitosterol and stigmasterol (5) [22], 1-monopalmitin (6) [23], β-sitosterol 3-O-β-glucopyranoside (daucosterol) (7) [24], 5-hydroxymethyl furfural (8) [25], ethyl β-glucopyranoside (9) [26], ethyl β-fructopyranoside (10) [25], kaempferol 3-O-β-glucopyranoside (11) [27], luteolin 7-O-β-glucopyranoside (12) [24], and apigenin 7-O-neohesperidoside (13) [28]. Among them, compounds 1–4, 6, 9, and 10 are first characterized herein in plants of the genus Ceiba. Moreover, compounds 1 and 4 have been previously detected by GC–MS in some species belonging to other plant families [29,30,31], whereas this is the first isolation of both molecules in the plant kingdom.

3.3. Anti-Infective Studies

Thus far, a number of research studies have deliberated the antimicrobial potential of different organic extracts of Chorisia species, namely Chorisia insignis H.B.K., Chorisia crispiflora H.B.K., Chorisia speciosa A. St.-Hil., Ceiba aesculifolia Kunth., and Ceiba pentandra L. against a variety of bacteria and fungi [12,13]. Nevertheless, the antimicrobial properties of these plants were not adequately described in the majority of the studies conducted so far via strict or more descriptive endpoints (e.g., MIC or IC₅₀ values), with only a few exceptions such as those reported by Franco et al. [32], Gomaa [33], and Behiry et al. [34], while in other works, relatively high concentrations of Chorisia extracts (in mg/mL) were used for testing of their antimicrobial potential and/or no appropriate controls were included [12,13]. Consequently, in the present work, the anti-infective potential of C. chodatii flowers was evaluated herein against a range of human pathogens following the standard guidelines provided by Cos et al. [7] for the screening of medicinal plants and their compounds against infectious organisms in an effort to search for potent anti-infective agents of natural origin with possible reasonable efficacy.

3.3.1. Antimicrobial Activity

Results of the antimicrobial assay revealed that none of the studied samples of C. chodatii flowers showed potent antimicrobial activities at the tested concentration ranges (8–200 µg/mL for the total extract and fractions and 0.8–20 µg/mL for the purified flavonoids). For all bacterial and fungal strains, the total extract of the flowers and its derived fractions showed IC₅₀ values higher than 200 µg/mL, whereas those of the tested compounds exceeded 20 µM (

Table 2. Anti-infective and anti-proliferative studies of C. chodatii flowers [IC₅₀ values (mean ± S.D.) in μg/mL for extracts and in µM for compounds and positive standards].

Sample	Antibacterial					Antifungal			Antileishmanial	Antimalarial		Anti-Proliferative
	MRSa	K. pneumoniae	E. coli	P. aeruroginosa	E. faecalis	A. fumigatus	C. neoformans	C. albicans	L. donovani	P. falciparum D6	P. falciparum W2	MDA-MB-468
Total flower extract	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	21.69 ± 0.03
Pet. ether fraction (I)	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	47.60 ± 0.09
Chloroform fraction (II)	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	25.46 ± 0.06
Ethyl acetate fraction (III)	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	27.39 ± 0.04
Aqueous fraction (IV)	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	>200	27.19 ± 0.03
Compound 11	>20	>20	>20	>20	>20	>20	>20	> 20	> 20	> 20	> 20	nt
Compound 12	>20	>20	>20	>20	>20	>20	>20	> 20	> 20	> 20	> 20	nt
Compound 13	>20	>20	>20	>20	>20	>20	>20	> 20	> 20	> 20	> 20	nt
Ciprofloxacin	0.23 ± 0.01	0.08 ± 0.02	0.01 ± 0.02	0.10 ± 0.01	0.43 ± 0.03	nt	nt	nt	nt	nt	nt	nt
Amphotericin B	nt	nt	nt	nt	nt	1.60 ± 0.02	1.08 ± 0.01	0.16 ± 0.01	nt	nt	nt	nt
Pentamidine	nt	nt	nt	nt	nt	nt	nt	nt	2.42 ± 0.02	nt	nt	nt
Chloroquine	nt	nt	nt	nt	nt	nt	nt	nt	nt	0.016 ± 0.01	0.155 ± 0.03	nt
Doxorubicin	nt	nt	nt	nt	nt	nt	nt	nt	nt	nt	nt	10.55 ± 0.03

). Amphotericin B and ciprofloxacin, on the other hand, displayed low IC₅₀ values that ranged between 0.16–1.60 and 0.01–0.23 µM against the tested fungal and bacterial species, respectively.

In this context, previous literature reports revealed the growth inhibitory potential of different solvent extracts of C. aesculifolia, C. pentandra, and C. speciosa against a variety of Gram-positive and Gram-negative bacteria as well as fungi, including S. aureus, E. faecalis, E. coli, K. pneumonia, P. aeruginosa, and C. albicans [35,36,37,38,39,40,41]. However, their antibacterial activities were reported at large concentrations, expressed via measurement of the diameter of inhibition zones only, and without comparison with suitable controls in most cases [12,13]. In contrast, Doughari and Ioryue [42] showed the inhibition of S. aureus, E. coli, P. aeruginosa, and Shigella dysentriae by the aqueous and ethanol extracts of C. pentandra stem bark, with minimum inhibitory and minimum bactericidal concentrations in the range of 6.25–50.00 mg/mL. Both extracts also displayed antifungal potential against C. albicans and Aspergillus flavus, with minimum fungicidal concentrations of 50–100 mg/mL [42]. Likewise, the alcohol and water extracts of C. pentandra were also active against C. albicans, Microsporum canis, Epidermophyton flocosum, and Trichopyton rubrum, showing MICs in the range of 50–100 mg/mL [43]. In another work, the methanol extract of C. aesculifolia fruits exerted antibacterial actions against S. aureus and E. faecalis, showing minimum inhibitory and minimum bactericidal concentrations of 2–14 mg/mL [32]. In the same framework, despite the broad-spectrum antimicrobial actions of luteolin, kaempferol, and apigenin derivatives [44,45,46], the weak antimicrobial activities observed herein for different extracts and isolated flavonoids of C. chodatii might be partly attributed to the low concentration ranges used in the assay [47].

3.3.2. Antileishmanial Activity

The obtained data indicated that both the studied fractions and flavonoids of C. chodatii flowers exerted low inhibitory effects against the growth of L. donovani promastigotes at the tested concentration ranges (0.8–20 µg/mL for the total extract and fractions and 0.4–10 µg/mL for the isolated flavonoids), showing IC₅₀ values above 200 µg/mL and 20 µM, respectively. In contrast, the IC₅₀ value of pentamidine was 2.42 µM (Table 2). Noteworthily, no previous reports have deliberated the antileishmanial potential of Ceiba plants. Furthermore, although some studies have indicated the antileishmanial properties of several flavonoids [48,49], the inffectiveness of both C. chodatii extracts and the isolated flavonoids might be due to the low concentration ranges used in the study [47].

3.3.3. Antimalarial Activity

Results of the in vitro antimalarial assay revealed that none of the tested strains of P. falciparum was sensitive to the studied samples of C. chodatii flowers, even at the highest tested concentration (15.87 µg/mL), with IC₅₀ values > 200 µg/mL recorded for the total extract and fractions and >20 µM for the pure compounds. On the contrary, chloroquine displayed potent inhibitory activities against both D6 and W2 strains (IC₅₀ = 0.016 and 0.155 µM, respectively) (Table 2). Based on the present literature data, this is the first testing of the antimalarial potential of Ceiba plants. While many plant flavonoids have shown good antiplasmodial properties [50], the weak activities exerted herein by different C. chodatii samples and flavonoids might be underlain by their small concentrations used in the study [47].

3.4. Anti-Proliferative Activity

Breast cancer is one of the most challenging malignancies and a major contributor to the global rates of cancer-related mortalities [51]. In this vein, TNBC exhibits the worst prognosis of all subtypes of breast cancer and accounts for about 15–20% of the identified cases. This awful disorder shows an earlier onset, more severe pathological profiles, and shorter survival rates [52]. Although both chemotherapy and radiotherapy are current therapeutic options, no fully effective or targeted treatments have been identified for TNBC thus far. Furthermore, these conventional treatment approaches are accompanied by serious side effects and their efficacy mostly diminishes when metastasis takes place [19].

Plants of the family Bombacaceae have been acknowledged as a source of secondary metabolites with promising cytotoxic potential against multiple cancer cells, e.g., breast cancer, including those of the genus Chorisia [12,13]. On this account, the inhibitory potential of the total extract of C. chodatii flowers and its derived fractions against the MDA-MB-468 TNBC (African American) cells was studied herein for the first time. The obtained data revealed the good anti-proliferative potential of the tested samples, of which the total flower extract exerted the highest inhibitory actions on the growth of MDA-MB-468 cells (IC₅₀ = 21.69 μg/mL), followed by the chloroform, aqueous, ethyl acetate, and petroleum ether fractions, with IC₅₀ values of 25.46, 27.19, 27.39, and 47.60 μg/mL, respectively. However, their inhibitory potential was lower than doxorubicin (IC₅₀ = 10.55 ± 0.03 μg/mL) in terms of IC₅₀ values (Table 2).

In consonance with the current findings, prior research studies have revealed the anti-proliferative potential of Ceiba species against a range of tumor cells [12,13]. Of these, the 70% ethanol extract of C. insignis leaves and its successive fractions showed significant inhibitory activities against the larynx cancer cell line HEP2, with IC₅₀ values ranging from 2.21 to 9.06 μg/mL compared with the anticancer agents cisplatin, doxorubicin, and 5-fluorouracil (IC₅₀ = 0.66, 0.74, and 2.2 μg/mL, respectively) [53]. However, lower anti-proliferative potential was reported for the same extracts against breast (MCF-7), colon (HCT116), liver (HepG2), and cervix (HELA) cancer cell lines [53]. Likewise, the n-hexane and ethyl acetate extracts of C. crispiflora leaves demonstrated potent inhibitory effects on MCF-7 cells via downregulation of NF-κB in a time- and concentration-dependent manner [54,55]. The methylene chloride fraction of the aerial parts of C. pentandra was also shown to exert prominent in vitro anti-proliferative actions against MCF-7 and HepG2 tumor cells (IC₅₀ = 18.859 and 14.895 µg/mL, respectively) [13,33], whereas the total extracts and different fractions of C. speciosa leaves exhibited much weaker cytotoxic effects against HepG2 cells, with IC₅₀ values of 57.30–954.99 µg/mL [41]. In view of this, despite showing moderate activities compared with doxorubicin, the observed effects of C. chodatii flowers against MDA-MB-468 TNBC cells could add to the current knowledge on the potential of these plants against breast cancer. Such effects of C. chodatii samples could be linked to their content of varied metabolites with reported anti-proliferative and anti-breast cancer properties, such as fatty acids and esters [56,57,58], monoglycerides [56,59,60], steroids [61,62], terpenoids [63,64], phenolic acids [65,66], anthocyanins [67,68], and flavonoids [62,69], including those isolated herein from the plant (e.g., compounds 2 [57], 5–8 [62,70,71,72,73], and 11–13 [74,75,76]), as well as their possible synergistic interactions [53,54,55,74].

4. Conclusions

The current work reported the effective use of the GC–MS-based metabolomics approach to unveil the lipophilic metabolites of C. chodatii flowers for the first time. A range of chemically diverse metabolites were therefore detected and characterized, of which long-chain aliphatic esters (77.016%) were the dominant group in the lipid fraction of the flowers. Further detailed phytochemical analysis of C. chodatii flowers using varied chromatographic and spectroscopic techniques led to the isolation and identification of 13 other metabolites of varied structural classes. On the other hand, different organic solvent extracts and the purified flavonoids from C. chodatii flowers did not display potent anti-infective aptitudes against a group of pathogens within concentration ranges up to 200 and 20 µg/mL, respectively. Taken together, the current findings generally add to our understanding of the metabolic capacity of Ceiba (Chorisia) plants and pave the way for future phytochemical exploration of various plant parts of C. chodatii trees. Additionally, the obtained data also threw up many questions in need of further investigation pertaining to the anti-infective potential of different members of the genus Ceiba.