ANTIMICROBIAL POTENTIAL OF CRUDE EXTRACTS AND FATTY ACID FRACTIONS OF FOUR PTERIDOPHYTE SPECIES FROM ASSAM, NORTHEAST INDIA, AND THEIR IDENTIFICATION BY GC-MS

ANTIMICROBIAL POTENTIAL OF CRUDE EXTRACTS AND FATTY ACID FRACTIONS OF FOUR PTERIDOPHYTE SPECIES FROM ASSAM, NORTHEAST INDIA, AND THEIR IDENTIFICATION BY GC-MS

S. Borkotoky * and V. V. Borah

Department of Biosciences, School of Life Sciences, Assam, Don Bosco University, Sonapur, Assam, India.

ABSTRACT: Fatty acids produced by plants protect themselves from pathogens, such as multidrug-resistant bacteria. In the present work, crude extracts, and fatty acid fractions of four pteridophyte species from Assam, Northeast India, were evaluated for their antimicrobial properties, and the bioactive compounds were identified using GC-MS analysis. The crude extract of Pteris semipinnata (Ps), Lygodium microphyllum (Lm), Lycodium flexuosum (Lf), and Lycopodiella cernua (Lc) was found to possess antibacterial and antifungal activities. The lowest MIC and MBC values were 0.25 mg/ml and 0.5 mg/ml for the fatty acid fractions against most of the microbial test strains. The GCMS analysis revealed 21 compounds in Ps, 35 in Lm, 33 in Lc, and 52 in Lf fractions. The present work supports the use of fern species in traditional medicine and therapies.

Keywords: Pteridophytes, Fatty acids, Antimicrobials, GC-MS

INTRODUCTION: About half of the deaths in tropical countries are caused by infectious diseases. Antibiotic resistance is a problem affecting the world and every nation. Antibiotic resistance occurs when bacteria become resistant and continue to proliferate in the presence of therapeutic amounts of an antibiotic, meaning the antibiotic can no longer effectively control or kill bacterial growth. Many factors contribute to antibiotic resistance. These include an inadequate grasp of how antibiotics function and how improper patient use promotes the development of resistance ¹. Drug-resistant bacteria have emerged because of the overuse of antibiotics in both medicine and food production.

Utilizing plant remedies is a straightforward solution to this issue. Herbal medications are intricate biological structures that have progressively evolved and contain hundreds of active substances that interact harmoniously. Long known for their antimicrobial effects, fatty acids (FA) are produced by plants to protect themselves from pathogens, such as multidrug-resistant bacteria ². Recently, FAs have also come to light as a possible antibiotic substitute.

Numerous FAs have been found to selectively inhibit various microbial pathogens, including Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens, Burkholderia cenocepacia, Vibrio spp., and Candida albicans suggesting their tremendous potential ³. Pteridophytes have several medicinal uses in folk medicine. In the tribal communities of Assam, pteridophytes have been used to treat various ailments, including colds and coughs, boils, cuts and wounds, respiratory problems, diarrhea, inflammation, bodily pain, hair loss and skin issues. From a single plant or a combination of plants, they create a paste, decoction, aqueous extract powder and juice ⁴. The majority of the tribes also manufacture their own native beer made from rice grains. Each tribe raises its unique starting culture for fermentation, made of different plant parts.

In the current work, crude extracts and fatty acids of four pteridophyte species used in starter culture for the Mishing tribe of Assam's rice beer preparation are evaluated for their antimicrobial properties. The bioactive compounds were identified using GC-MS analysis.

MATERIALS AND METHOD:

Collection of Samples: The selected plant specimens were – Pteris semipinnata, Lygodium microphyllum, Lygodium flexuosum, and Lycopodiella cernua. Fresh plants were collected in the month of March from a Mishing village in Jorhat, Assam (26º 46’ 33.012” N Latitude and 94º 15’ 29.772” E Longitude). The leaves of Pteris semipinnata (Ps), Lygodium microphyllum (Lm), Lygodium flexuosum (Lf) and leaves along with stem of Lycopodiella cernua (Lc) were collected for the study. The herbarium specimens were prepared and identified in the Weed Herbarium of Assam Agricultural University. Plant material was collected, washed, shade dried, powdered with a blender, and stored in airtight bottles.

Preparation of Extracts: Five grams of each plant powder was extracted separately using different solvents based on polarity (ethyl acetate< acetone< ethanol< methanol) in the Soxhlet apparatus for 6 hours. The crude extracts were evaporated in a rotary vacuum evaporator at 37 ºC and stored at 4 ºC.

Extraction of Fatty Acids: 2 gm of aqueous extract was treated with 10 % lead acetate solution. The supernatant was collected, diluted with water and acidified with 1 % HCl before boiling for 2-3 hours. The precipitate was then extracted with ethanol and purified by fractional crystallization to obtain the fractions Ps-F, Lm-F, Lf-F and Lc-F. These fractions were then subjected to antimicrobial screening against clinical isolates and reference strains to establish their antimicrobial potential ⁵.

Antimicrobial Activity:

Microorganisms: The microorganisms selected for this study were isolated from clinical samples collected from Ayursundra Superspeciality Hospital, Guwahati, Assam. These included Pseudomonas aeruginosa, Salmonella typhi, Klebsiella pneumoniae, Acinetoba cterbaumannii, Proteus mirabilis, Candida albicans, and C. tropicalis. The reference strains used in this study are Staphylococcus epidermidis ATCC 35984 and K. pneumoniae ATCC BAA-1705. Cultures were maintained on Luria Bertani Agar (for bacteria) and Potato Dextrose Agar (for yeast) at 4°C.

Determination of MIC and MBC: MIC and MBC were determined using resazurin microtiter assay. Two-fold serial dilutions of the crude extracts and fractionated extracts were made directly in a microtiter plate with Mueller Hinton broth (MHB) to produce varying concentrations. MHB was used to adjust microbial cultures to 0.5 McFarland turbidity standard (1.5 × 10⁸ CFU/ml). Amoxicillin was used as a standard for positive control. The plate was sealed with a sterile sealer and kept at 37°C for 24 hours. After incubation, 0.02 % resazurin (Himedia-RM125-1G) was added to each microtiter plate well and incubated for 30 minutes at 37°C. The wells that had microbial growth became pink, whereas the wells that did not have microbial growth remained blue. The wells corresponding to the MIC and with higher concentration were streaked onto Mueller-Hinton Agar plates and incubated overnight at 37°C. The concentration corresponding to no bacterial growth was recorded as the minimum bactericidal concentration (MBC) for extract ⁶.

Identification of the Fatty Acid Fraction with Gas Chromatography and Mass Spectroscopy: The fatty acid fractions were re-dissolved in spectroscopy grade ethanol and filtered through 0.2 mm filter for GCMS analysis performed in a Perkin Elmer (USA) Clarus 680/600C unit fitted with Elite 5 MS column (length: 30 m, ID: 0.25 mm, film thickness: 0.25 mm). The software used in the system is TurboMassver 5.4.2. The oven program started at 60°C for 1 min and ramped at 7°C/min up to 200°C and held for 3 mins, again ramped at 10°C/min to 300°C and then held for 5 min. Next, 1.0 ml sample was injected at 280°C using He as carrier gas with a solvent delay of 5 min. The split ratio was 10: 1. The mass spectrometer (Clarus 600C; single quad) was operated in the electron ionization (EI) mode at 70 eV with a source temperature of 150°C and a continuous scan from m/z 50 to 600. The peaks were identified by matching the mass spectra with the National Institute of Standards and Technology (NIST) library, USA.

RESULTS AND DISCUSSION: The development of new drugs also depends on the study of medicinal plants. Most underdeveloped nations presently use herbal treatment, and wealthy countries are also quickly adopting it. The pharmacological effects of plants are caused by metabolites, which are organic compounds divided into primary and secondary metabolites. Plants produce secondary metabolites such as alkaloids, flavonoids, saponins, terpenoids, steroids, glycosides, tannins, volatile oils, and other substances to protect themselves from microbial infections and insect invasions⁷.

The experiment to ascertain the antimicrobial efficacy of the different crude extracts and fatty acid fraction utilized seven clinical and two control isolates. The antimicrobial activity of the different extracts was effective against all of the test microorganisms at dosages ranging from 0.29 to 17.75 mg/ml, as shown in Table 1-3. Fractionation sometimes leads to improved biological activity, as seen in this study. The lowest MIC and MBC values were 0.25 mg/ml and 0.5 mg/ml, respectively Table 1-3, recorded for the fatty acid fractions Ps-F, Lc-F, Lf-F, and Lm-F against most of the microbial test strains. The low MIC observed in this study further supports the potential of fatty acid as an antimicrobial resource. The results align with earlier research on the antimicrobial effectiveness of fatty acids against related species employed in this study ^{3, 8-9}.

TABLE 1: MIC AND MBC OF THE DIFFERENT CRUDE EXTRACTS AND FRACTIONATED EXTRACTS AGAINST BACTERIAL CLINICAL ISOLATES

TABLE 2: MIC AND MBC OF THE DIFFERENT CRUDE EXTRACTS AND FRACTIONATED EXTRACTS AGAINST FUNGAL CLINICAL ISOLATES

TABLE 3: MIC AND MBC OF THE DIFFERENT CRUDE EXTRACTS AND FRACTIONATED EXTRACTSAGAINSTBACTERIAL CONTROL ISOLATES

Based on the promising result of the antimicrobial study, the fatty acid fractions were further subjected to GCMS analysis to identify the bioactive compounds. The four pteridophytes' fatty acid fraction included 184 peaks with retention times ranging from 6.044 to 38.157 Fig. 1-4. There were 21 compounds found in the Pterissemipinnata fraction, 35 compounds in the Lygodium flexuosum fraction, 33 compounds in the Lycopodiella cernua fraction, and 52 compounds in the Lygodium microphyllum fraction after a NIST library search was conducted for the significant peaks Table 4-7.

FIG. 1: GC-MS CHROMATOGRAM OF THE PTERISSEMIPINATTA FRACTION

TABLE 4: GC-MS ANALYSIS OF FRACTIONATED EXTRACT OF PTERIS SEMIPINNATA

Sl. no.	Compound name	Molecular structure	Mol formula	Mol.wt	Activity
1	3-methyl-2-(2-oxopropyl)furan		C8H10O2	138	Antimicrobial activity10-11, Antibiofilm,  Anti-quorum sensing activity12; antioxidant, antipyretic, anti-inflammatory activity13
2	z,z-6,28-heptatriactontadien-2-one		C37H70O	530	Larvicidal activity14; Vasodialator, carcinogenic and antioxidant activity15
3	z,z-6,27-hexatriactontadien-2-one		C36H68O	516	Vasodialator16
4	11,14-eicosadienoic acid, methyl ester		C21H38O2	322	Anti-infammatory, anti-oxidant, anti-arthritic, anticoronary17
5	Undec-10-ynoic acid, tetradecyl ester		C25H46O2	378	Antimicrobial activity18
6	7-hexadecenal, (z)-		C16H30O	238	-
7	Undec-10-ynoic acid, octadecyl ester		C29H54O2	434	Inhibitor of cytochrome P450 4A119; Antioxidant, Antifungal, and Wound Healing Activity20
8	Tridecane, 6-cyclohexyl-		C19H38	266	-
9	Neophytadiene		C20H38	278	Anti-inflammatory and antiviral activity21; Antioxidant22
10	Undecane, 3-cyclohexyl-		C17H34	238	-
11	Tridecane, 3-cyclohexyl-		C19H38	266	-
12	1,2-15,16-diepoxyhexadecane		C16H30O2	254	Cytotoxic activity23; Anti-tumour and anti-inflammatory activity24
13	Docosanal		C22H44O	324	Antiviral activity25
14	Dimethylsulfoxonium formylmethylide		C4H8O2S	120	Antioxidant and Cytotoxic activity26; Antimicrobial activity27
15	Ethane, 1,1-dichloro-		C2H4Cl2	98	-
16	2-chloroethyl methyl sulfone		C3H7ClO2S	142	-
17	(z)-1-chloro-2-(methylsulfonyl)ethylene		C3H5ClO2S	140	-
18	2-chloropropionyl chloride		C3H4Cl2O	126	-
19	Propane, 1,2-dichloro-		C3H6Cl2	112	-
20	Disilane, 1,1,2,2-tetrachloro-1,2-dimethyl-		C2H6Cl4Si2	226	-
21	Bis(methylsulfonyl)methane		C3H8O4S2	172	Anti-inflammatory activity 28

*Molecular structures were generated using ChemDraw Ultra 12.0

FIG. 2: GC-MS CHROMATOGRAM OF THE LYGODIUM FLEXUOSUM FRACTION

TABLE 5: GC-MS ANALYSIS OF FRACTIONATED EXTRACT OF LYGODIUM FLEXUOSUM

Sl. no.	Compound name	Molecular structure	Mol formula	Mol. wt	Activity
1	Hexacosyl acetate		C28H56O2	424	Larvicidal activity29; Antifungal activity30
2	Chloroacetic acid, tetradecyl ester		C16H31ClO2	290	Antioxidant, antimicrobial and bacteriocide, anti- infammatory activity13
3	1-hexadecanol		C16H34O	242	Antimicrobial activity31
4	Cis-1-chloro-9-octadecene		C18H35Cl	286	Antibacterial activity32
5	Acetic acid, chloro-, hexadecyl ester		C18H35ClO2	318	antibacterial, anthelmintic, insecticidal activity33
6	N-nonadecanol-1		C19H40O	284	Antibacterial activity34
7	Behenic alcohol		C22H46O	326	Antiviral activity35
8	Octacosanol		C28H58O	410	Antinociceptive and Anti-Inflammatory activity36
9	1-heneicosanol		C21H44O	312	Antioxidant and Antimicrobial activity37
10	1-heneicosyl formate		C22H44O2	340	Biocontrol activity38
11	1-hentetracontanol		C41H84O	593	Antimicrobial activity39
12	trifluoroacetic acid, pentadecyl ester		C17H31F3O2	324	-
13	Disparlure		C19H38O	282	-
14	Dotriacontyl pentafluoropropionate		C35H65F5O2	612	Cytotoxic activity40
15	Heptafluorobutyric acid, n-tetradecyl ester		C18H29F7O2	410	Antioxidant activity41
16	Benzene propanoic acid, 3,5-bis(1,1-dimethylethyl)-4-hydroxy-, methyl e		C18H28O3	292	Antibacterial activity42
17	4-(3,5-di-tert-butyl-4-hydroxyphenyl)butyl acrylate		C21H32O3	332	-
18	bis[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propyl] maleate		C38H56O6	608	Antioxidant activity43
19	butylated hydroxytoluene, tms derivative		C18H32OSi	292	Antioxidant activity44
20	3-(3,5-di-tert-butyl-4-hydroxyphenyl)propyl methacrylate		C21H32O3	332	-
21	Eicosen-1-ol, cis-9-		C20H40O	296	Antibacterial activity45
22	9-octadecen-1-ol, (z)-		C18H36O	268	-
23	13-docosen-1-ol, (z)-		C22H44O	324	-
24	z,z-6,28-heptatriactontadien-2-one		C37H70O	530	Larvicidal activity46
25	1,16-hexadecanediol		C16H34O2	258	-
26	(z)-14-tricosenyl formate		C24H46O2	366	Anticancer activity47
27	1,19-eicosadiene		C20H38	278	Antimicrobial activity48; Anti-quorum sensing and Anti-biofilm activity49
28	Oleyl alcohol, trifluoroacetate		C20H35F3O2	364	Antioxidant, Antidiabetic and Hypolipidemic50
29	Ethanol, 2-(9-octadecenyloxy)-, (z)-		C20H40O2	312	Antimycotoxigenic activity51
30	Cyclododecanemethanol		C13H26O	198	Antioxidant and Metal Chelation Activity52
31	11-dodecen-1-ol trifluoroacetate		C14H23F3O2	280	Antiobesity and Antihyperlipidemic activity53
32	9-hexadecenoic acid, 9-octadecenyl ester, (z,z)-		C34H64O2	504	Antibacterial activity54
33	17-octadecynoic acid		C18H32O2	280	-
34	z,e-2-methyl-3,13-octadecadien-1-ol		C19H36O	280	-
35	1,21-docosadiene		C22H42	306	-

*Molecular structures were generated using ChemDraw Ultra 12.0

FIG. 3: GC-MS CHROMATOGRAM OF THE LYCOPODIELLA CERNUA FRACTION

TABLE 6: GC-MS ANALYSIS OF FRACTIONATED EXTRACT OF LYCOPODIELLA CERNUA

Sl. no.	Compound name	Molecular structure	Mol formula	Mol. wt	Activity
1	4-n-hexylthiane, s,s-dioxide		C11H22O2S	218	-
2	oleic acid		C18H34O2	282	Antimicrobial, Antifungal, anticonvulsive activity, Antiadhesive, Antiallergic, Antianalgesic, Antiatherosclerosis, Anesthetic, Antihelmenthic, Antianxiety, Antibacterial, Antiberiberi, Antibiotic, Anticancer, Anticonvulsant, Antidiabetic, Antidiarrheic, Antifertility, Antigastric, Anti-infammatory, Antiobesity, Antioxidant, Antiulcer, Antitubercellosic, Anticold, Antihepatotoxic and Antiviral activityanemiagenic, dermatitigenic13
3	2-nonadecanone 2,4-dinitrophenylhydrazine		C25H42N4O4	462	Antioxidant activity55
4	Pentadecanoic acid, 14-bromo-		C15H29BrO2	320	-
5	Docosanoic acid		C22H44O2	340	Antibacterial activity56
6	Tetracosanoic acid, isobutyl ester		C28H56O2	424	-
7	Docosanoic acid, docosyl ester		C44H88O2	648	Antidepressant and Cytotoxic activity56
8	Butyl 11-eicosenoate		C24H46O2	366	-
9	Eicosanoic acid		C20H40O2	312	Reduced heart diseases, kidney and liver function, blood Clotting13; Antibacterial and cytoprotective activity57
10	Propionic acid, 3-iodo-, octadecyl ester		C21H41IO2	452	-
11	Tetracosanoic acid		C24H48O2	368	-
12	l-(+)-ascorbic acid 2,6-dihexadecanoate		C38H68O8	652	Antioxidant and reduces the triglycerides level—Protects LDL against peroxidation and inhibits the progression of atherosclerosis, Antiallergic, Antianemic, Antianxiety, Antibacterial, Antibronchitic, Anticancer, Anticarcinogenic, Anticataract, Anticoagulant, Anticonvulsant, Antidiabetic, Antidiarrheic, Antifatigue, Antifertility, Antigastric, Anti-infammatory, Antimalarial, Antioxidant, Antistress, Antiulcer, Antiatheroscelerotic, Anticold, Antiglaucomic, Antihepatic, Antihypertensive, Antiplague, Antiproliferant, Antiprotozoal, Antiseptic, Antistroke, Antituberculic, Antitumer, CNSStimulant, Chelator, Chemopreventive, CytochromeP450Inducer, Deodorant, Dermal, Detoxicant, Flavor, Hypolipidimic, Neuroprotective, Neurotransmitter, Termiticide and Antiviral activity13
13	Octadecanoic acid		C18H36O2	284	Decreases cardiovascular and cancer risks, reduces LDL cholesterol levels, reduces blood pressure, improved heart function58
14	Cyclohexane, 1-(1-tetradecylpentadecyl)-		C35H70	490	-
15	i-propyl 10-methyl-dodecanoate		C16H32O2	256	-
16	Butyl 15-methylhexadecanoate		C21H42O2	326	-
17	Docosanoic acid, isobutyl ester		C26H52O2	396	-
18	9,9-dimethoxybicyclo[3.3.1]nona-2,4-dione		C11H16O4	212	-
19	Cis-2-methyl-4-n-pentylthiane, s,s-dioxide		C11H22O2S	218	Anti-proliferative activity59
20	Chloroacetic acid, tetradecyl ester		C16H31ClO2	290	Antioxidant, antimicrobial and bacteriocide, anti- infammatory activity13
21	5-methyl-z-5-docosene		C23H46	322	Antibacterial, antidiabetic, antitumour activities13
22	Heptyl triacontyl ether		C37H76O	536	-
23	Chloroacetic acid, dodecyl ester		C14H27ClO2	262	-
24	Eicosyl heptyl ether		C27H56O	396	-
25	Heptyl hexacosyl ether		C33H68O	480	-
26	Trans-2,4-dimethylthiane, s,s-dioxide		C7H14O2S	162	Anti-infammatory activity60
27	Eicosyl nonyl ether		C29H60O	424	Antioxidant activity61
28	triarachine		C63H122O6	975	Anti-hyperglycemic activity62
29	3-methyl-2-(2-oxopropyl) furan		C8H10O2	138	Antioxidant, antimicrobial and bacteriocide, Antipyretic, antiinfammatory activity13
30	z,z-6,27-hexatriactontadien-2-one		C36H68O	516	Vasodilator16
31	Heptacosanoic acid, 25-methy—, methyl ester		C29H58O2	438	Antimicrobial activity13
32	11,14-eicosadienoic acid, methyl ester		C21H38O2	322	Anti-infammatory, anti-oxidant, anti-arthritic, anticoronary13
33	1,3-dioxolane, 4-ethyl-5-octyl-2,2-bis(trifluoromethyl)-, trans-		C15H24F6O2	350	Antioxidant activity63

*Molecular structures were generated using ChemDraw Ultra 12.0

FIG. 4: GC-MS CHROMATOGRAM OF THE LYGODIUM MICROPHYLLUM FRACTION

TABLE 7: GC-MS ANALYSIS OF FRACTIONATED EXTRACT OF LYGODIUM MICROPHYLLUM

Sl. no.	Compound name	Molecular structure	Mol formula	Mol.wt	Activity
1	Oleic acid		C18H34O2	282	Antimicrobial, Antifungal, anticonvulsive activity, Antiadhesive, Antiallergic, Antianalgesic, Antiatherosclerosis, Anesthetic, Antihelmenthic, Antianxiety, Antibacterial, Antiberiberi, Antibiotic, Anticancer, Anticonvulsant, Antidiabetic, Antidiarrheic, Antifertility, Antigastric, Anti-infammatory, Antiobesity, Antioxidant, Antiulcer, Antitubercellosic, Anticold, Antihepatotoxic and Antiviral activityanemiagenic, dermatitigenic13
2	Docosanoic acid		C22H44O2	340	Antibacterial and cytoprotective activity57
3	Eicosanoic acid		C20H40O2	312	Reduced heart diseases, kidney and liver function, blood Clotting13; Antibacterial and cytoprotective activity57
4	Tetracosanoic acid		C24H48O2	368	-
5	Propionic acid, 3-iodo-, octadecyl ester		C21H41IO2	452	-
6	Octadecanoic acid		C18H36O2	284	Decreases cardiovascular and cancer risks, reduces LDL cholesterol levels, reduces blood pressure, improved heart function58
7	Pentadecanoic acid, 14-bromo-		C15H29BrO2	320	-
8	1,3-dioxolane, 4-ethyl-5-octyl-2,2-bis(trifluoromethyl)-, trans-		C15H24F6O2	350	Antioxidant activity63
9	methyl 2-hydroxy-eicosanoate		C21H42O3	342	-
10	2-nonadecanone 2,4—dinitrophenylhydrazine		C25H42N4O4	462	Antimicrobial activity15
11	d-mannitol, 1-o-(22-hydroxydocosyl)-		C28H58O7	506	-
12	Distearyl thiodipropionate		C42H82O4S	682	Antioxidant activity64
13	l-(+)-ascorbic acid 2,6-dihexadecanoate		C38H68O8	652	Antioxidant and reduces the triglycerides level—Protects LDL against peroxidation and inhibits the progression of atherosclerosis, Antiallergic, Antianemic, Antianxiety, Antibacterial, Antibronchitic, Anticancer, Anticarcinogenic, Anticataract, Anticoagulant, Anticonvulsant, Antidiabetic, Antidiarrheic, Antifatigue, Antifertility, Antigastric, Anti-infammatory, Antimalarial, Antioxidant, Antistress, Antiulcer, Antiatheroscelerotic, Anticold, Antiglaucomic, Antihepatic, Antihypertensive, Antiplague, Antiproliferant, Antiprotozoal, Antiseptic, Antistroke, Antituberculic, Antitumer, CNSStimulant, Chelator, Chemopreventive, CytochromeP450Inducer, Deodorant, Dermal, Detoxicant, Flavor, Hypolipidimic, Neuroprotective, Neurotransmitter, Termiticide and Antiviral activity13
14	Decanoic acid, silver(1+) salt		C10H19AgO2	278	-
15	Tetradecanoic acid		C14H28O2	228	Anti-virulence activity65
16	Pentadecanoic acid		C15H30O2	242	Antibacterial Antifungal activity66
17	9-oxononanoic acid		C9H16O3	172	-
18	9,9-dimethoxybicyclo[3.3.1]nona-2,4-dione		C11H16O4	212	antioxidants,  anti-arthritic  and  antimicro-bial activity67
19	i-propyl 10-methyl-dodecanoate		C16H32O2	256	-
20	Strychane, 1-acetyl-20.alpha.-hydroxy-16-methylene-		C21H26N2O2	338	Antimicrobial activity68
21	n-hexadecanoic acid		C16H32O2	256	Anti-inflammatory activity69; Antioxidant, hypocholesterolemic, nematicide, pesticide, antiandrogenic, flavour, hemolytic, 5-alpha reductase inhibitor70
22	Ethyl 14-methyl-hexadecanoate		C19H38O2	298	Insecticidal and Anti-helminthic activity71
23	octadecanoic acid, 17-methyl-, methyl ester		C20H40O2	312	-
24	10-bromodecanoic acid, ethyl ester		C12H23BrO2	278	Antioxidant and Antibacterial activity72
25	nonanoic acid, 9-bromo-, ethyl ester		C11H21BrO2	265	Antioxidant and Antimicrobial activity73
26	Octadecanoic acid, ethyl ester		C20H40O2	312	Antimicrobial activity74
27	Hexadecanoic acid, ethyl ester			284	Antioxidant, hypocholesterolemic, nematicide, pesticide, antiandrogenic, flavor, hemolytic, 5-alpha reductase inhibitor71
28	Ethyl 13-methyl-tetradecanoate		C17H34O2	270	Antimicrobial, Antioxidant and Anticancer activity75
29	methyl 19-methyl-eicosanoate		C22H44O2	340	-
30	Tetradecanoic acid, ethyl ester		C16H32O2	256	Cytotoxic activity76
31	Docosanoic acid, docosyl ester		C44H88O2	648	Antidepressant and Cytotoxic activity56
32	Dotriacontyl isopropyl ether		C35H72O	508	-
33	cis-2-methyl-4-n-pentylthiane, s,s-dioxide		C11H22O2S	218	Anti-proliferative activity57
34	5-methyl-z-5-docosene		C23H46	322	Antibacterial, antidiabetic, antitumour activities13
35	Chloroacetic acid, tetradecyl ester		C16H31ClO2	290	Antioxidant, antimicrobial and bacteriocide, anti- infammatoryactivity13
36	triarachine		C63H122O6	975	Anti-hyperglycemic activity62
37	Trans-2,4-dimethylthiane, s,s-dioxide		C7H14O2S	162	Anti-infammatory activity60
38	Eicosyl nonyl ether		C29H60O	424	Antioxidant activity61
39	6,10-dimethyl-4-undecanol		C13H28O	200	Antioxidant And Anti-cancer activity77
40	Trans-2-methyl-4-n-pentylthiane, s,s-dioxide		C11H22O2S	218	Antioxidant activity63
41	Trans-2-methyl-4-n-butylthiane, s,s-dioxide		C10H20O2S	204	Antimicrobial, Antioxidant, cytotoxic activity78; Anti-proliferative and apoptosis-inducing activity79
42	Dodecane, 1-fluoro-		C12H25F	188	Antioxidant activity80
43	Butyl 9-hexadecenoate		C20H38O2	310	-
44	cis-1-chloro-9-octadecene		C18H35Cl	286	Antiviral activity81
45	Undec-10-ynoic acid, tetradecyl ester		C25H46O2	378	Antimicrobial activity18
46	9-octadecenoic acid, 1,2,3-propanetriyl ester, (e,e,e)-		C57H104O6	884	Antispasmodic and immune modulators82
47	z,z-6,28-heptatriactontadien-2-one		C37H70O	530	Vasodilator13
48	oxirane, tetradecyl-		C16H32O	240	Antimicrobial activity83
49	3-methyl-2-(2-oxopropyl)furan		C8H10O2	138	Antimicrobial activity10-11, Antibiofilm,  Anti-quorum sensing activity12; antioxidant, antipyretic, anti-inflammatory activity13
50	2-methyltetracosane		C25H52	352	Antioxidant Activity84
51	1-nonylcycloheptane		C16H32	224	Antimicrobial activity85
52	1,10-hexadecanediol		C16H34O2	258	Antioxidant and Antibacterial activity86

*Molecular structures were generated using ChemDraw Ultra 12.0

The various medicinal uses of pteridophytes ^87-88 may be due to the bioactive chemicals found in the fractions, which have been documented to exhibit a variety of medicinal properties Table 4-7. With the existence of these bioactive chemicals, the MIC and MBC values of the pteridophyte extracts in the current investigation may also be confirmed. To have a better understanding of the therapeutic activities of the examined pteridophytes, it will be interesting to document the biological roles of some of the compounds present in greater quantities, such as Tridecane, 6-Cyclohexyl-, Undecane, 3-Cyclohexyl-, Ethane, 1,1-Dichloro-, 2-Chloroethyl Methyl Sulfone, (Z)-1-Chloro-2-(Methylsulfonyl) Ethylene, 2-Chloropropionyl Chloride, Propane, 1,2-Dichloro-, Disilane, 1,1,2,2-Tetrachloro-1,2-Dimethyl-, Disparlure, 4-(3,5-Di-Tert-Butyl-4-Hydroxyphenyl)Butyl Acrylate, 3-(3, 5-Di-Tert-Butyl – 4 - Hydroxyphenyl) Propyl Methacrylate, 9-Octadecen-1-Ol, (Z)-, 1,16-Hexadecanediol, 17-Octadecynoic Acid, Z,E-2-Methyl - 3, 13 – Octadecadien – 1 - Ol, 1, 21-Docosadiene, Pentadecanoic Acid, 14-Bromo-, Tetracosanoic Acid, D-Mannitol, 1-O-(22-Hydroxydocosyl)- and Decanoic Acid, Silver(1+) Salt, which has not yet been published.

CONCLUSION: The present work found that the four pteridophyte species possess antibacterial and antifungal activities. The study revealed encouraging outcomes for fatty acids as antimicrobial agents.

The GCMS analysis of the fatty acid fractions revealed bioactive compounds with reported biological activity, which supports the study's results. The present work supports the use of fern species in traditional medicine and therapies. These results will also be useful for undertaking in-silico and cell-line studies for potential pharmacological lead compounds for drug discovery in future research.

ACKNOWLEDGMENTS: The authors are grateful to Dr. Juri Bharat Kalita and Dr. Himadri Dutta from the Microbiology Department of Ayursundra superspeciality Hospital for supplying the clinical microbial isolates for antimicrobial studies. The authors are also grateful to Dr. Rajiv Chandra Goswami, Guwahati Biotech Park, for the guidance and information on GC-MS analysis.

CONFLICTS OF INTEREST: The authors (S. Borkotoky and V. V. Borah) have no conflicts of interest to declare.

REFERENCES:

1. Tillotson G: The Fight against Bacterial Resistance – New Initiatives But Much Still Needed.Journal of Infectious Diseases and Therapy 2016. doi:10.4172/2332-0877.1000e109.
2. Giancarlo CV, Carlimar OM, Solymar M, Christian MG, Valle RGD, Carballeira N and Sanabria-Ríos DJ: Antibacterial fatty acids: An update of possible mechanisms of action and implications in the development of the next-generation of antibacterial agents, Progress in Lipid Research 2021; 82: 101093, https://doi.org/10.1016/j.plipres.2021.101093.
3. Kumar P, Lee JH and Lee HBJ: Fatty Acids as Antibiofilm and Antivirulence Agents, Trends in Microbiology, 2020; 28(9): 753-768.
4. Sen A and Ghosh PD: A note on the ethnobotanical studies of some pteridophytes in Assam, IJTK 2011; 10(2): 292-295.
5. Twitchell E: The Precipitation of Solid Fatty Acids with Lead Acetate in Alcoholic Solution. Ind Eng Chem 1921; 13(9): 806–807 https://doi.org/10.1021/ie50141a024
6. Khanal L, Sharma K, Pokharel Y and Kalauni S: Assessment of Phytochemical, Antioxidant and Antimicrobial Activities of Some Medicinal Plants from Kaski District of Nepal. American Journal of Plant Sciences 2020; 11: 1383-1397.
7. Antonisamy MA, Johnson & Johnson, Antonisamy, Madona C, Ray A, Cruz-Martins N and Coutinho H: Antibiotics in-vitro toxicity, antioxidant, anti-inflammatory, and antidiabetic potential of Sphaerostephanos unitus (L.) Holttum. Antibiotics 2020; 9: 333. 10.doi: 10.3390/antibiotics9060333
8. Desbois A: Potential applications of antimicrobial fatty acids in medicine, agriculture and other industries. Recent Pat Antiinfect Drug Discov 2012; 7(2): 111–22.
9. Yoon B, Jackman J, Valle-Gonz´ alez E and Cho N: Antibacterial free fatty acids and monoglycerides: biological activities, experimental testing, and therapeutic applications. Int J Mol Sci 2018; 19(4): 1114.
10. Singh N, Sudandiradoss C and Abraham J: Screening of furanone in cucurbitamelo and evaluation of its bioactive potential using in-silico Interdiscip Sci Comput Life Sci 2016; 8: 395–402. https://doi.org/10.1007/s12539-016-0161-z
11. Karthik R, Saravanan R and Ebenezar KK: Isolation, Purification, and Characterization of Avian Antimicrobial Glycopeptide from the Posterior Salivary Gland of Sepia pharaonis. Appl Biochem Biotechnol 2015; 175: 1507–1518. https://doi.org/10.1007/s12010-014-1370-8
12. Gupta K, Singh SP and Manhar AK: Inhibition of Staphylococcus aureus and Pseudomonas aeruginosa Biofilm and Virulence by Active Fraction of Syzygium cumini (L.) Skeels Leaf Extract: In-Vitro and In-Silico Studies. Indian J Microbiol 2019; 59: 13–21. https://doi.org/10.1007/s12088-018-0770-9
13. Ralte L, Khiangte L and Thangjam NM: GC–MS and molecular docking analyses of phytochemicals from the underutilized plant, Parkiatimoriana revealed candidate anti-cancerous and anti-inflammatory agents. Sci Rep 2022; 12: 3395. https://doi.org/10.1038/s41598-022-07320-2
14. Manjari MS, Karthi S, Ramkumar G, Muthusamy R, Natarajan D and Shivakumar MS: Chemical composition and larvicidal activity of plant extracts from Clausena dentata (Willd) (Rutaceae) against dengue, malaria, and filariasis vectors. Parasitol Res 2014; 113(7): 2475-81. doi: 10.1007/s00436-014-3896-7. Epub 2014 May 7. PMID: 24802866.
15. Ramkumar G, Karthi S, Muthusamy R, Suganya P, Natarajan D, Kweka J and Eliningaya: Chemical composition of chloroform leaf extract from Glycosmis pentaphylla. PLOS ONE. Dataset. 2016https://doi.org/10.1371/journal.pone.0158088.t004
16. Mallikadevi T, Paulsamy S, Jamuna S and Karthika K: Asian J Pharm Clinical Res., 2012, 5(4), 163-168.
17. Nithyadevi J and Sivakumar R: Phytochemical screening and GC-MS, FT-IR analysis of methanolic extract leaves of Solanumtorvum Sw. Int J Res Stud Bio 2015; 3: 61–66.
18. Hasan MD, Basher M, Shyama M, Fatema J, Islam MD and Sarmina Y: Antimicrobial activities and GC-MS chemical profiling of methanol extracts of different parts of Morus alba (Tut) in Bangladesh. IJB 2021; 18: 156-166. 10.12692/ijb/18.5.156-166.
19. Khiralla A, Spina R, Varbanov M, Philippot S, Lemiere P, Slezack-Deschaumes S, André P, Mohamed I, Yagi SM and Laurain-Mattar D: Evaluation of Antiviral, Antibacterial and Antiproliferative Activities of the Endophytic Fungus Curvularia papendorfii, and Isolation of a New Polyhydroxyacid. Microorganisms 2020; 8(9): 1353. https://doi.org/10.3390/microorganisms8091353
20. Abdel-Motaal FF, Maher ZM, Ibrahim SF, El-Mleeh A, Behery M and Metwally AA: Comparative Studies on the Antioxidant, Antifungal, and Wound Healing Activities of Solenostemmaarghel Ethyl Acetate and Methanolic Extracts. Applied Sciences 2022; 12(9): 4121. https://doi.org/10.3390/app12094121
21. Ratheesh M, Sunil S and Sheethal S: Anti-inflammatory and anti-COVID-19 effect of a novel polyherbal formulation (Imusil) via modulating oxidative stress, inflammatory mediators and cytokine storm. Inflammopharmacol 2022; 30: 173–184. https://doi.org/10.1007/s10787-021-00911-x
22. Bhardwaj M, Sali VK and Mani S: Neophytadiene from Turbinariaornata Suppresses LPS-Induced Inflammatory Response in RAW 264.7 Macrophages and Sprague Dawley Rats. Inflammation 2020; 43: 937–950. https://doi.org/10.1007/s10753-020-01179-z
23. Ganesh M and Mohankumar M: Extraction and identification of bioactive components in Sidacordata (Burm.f.) using gas chromatography-mass spectrometry. J Food Sci Technol 2017; 54(10): 3082-3091. doi: 10.1007/s13197-017-2744-z. Epub 2017 Jul 17. PMID: 28974793; PMCID: PMC5602971.
24. Shareef HK, Muhammed HJ, Hussein HM and Hameed IH: Antibacterial effect of ginger (Zingiber officinale) roscoe and bioactive chemical analysis using gas chromatography mass spectrum. Oriental Journal of Chemistry 2016; 32(2): 817-837. Doi: 10.13005/ojc/320207
25. Shankar VK, Wang M, Ajjarapu S, Kolimi P, Avula B, Murthy R, Khan I and Murthy SM: Analysis of docosanol using GC/MS: Method development, validation and application to ex vivo human skin permeation studies. Journal of Pharmaceutical Analysis 2022; 12(2): 287-292. https://doi.org/10.1016/j.jpha.2021.08.004.
26. Paudel MR, Chand MB, Pant B and Pant B: Antioxidant and cytotoxic activities of Dendrobium moniliforme extracts and the detection of related compounds by GC-MS. BMC Complement Altern Med 2018; 18(1): 134. doi: 10.1186/s12906-018-2197-6. PMID: 29685150; PMCID: PMC5913799.
27. Paudel MR, Chand MB, Pant B and Pant B: Assessment of Antioxidant and Cytotoxic Activities of Extracts of Dendrobium crepidatum. Biomolecules 2019; 9(9): 478. https://doi.org/10.3390/biom9090478
28. Butawan M, Benjamin RL and Bloomer RJ: Methylsulfonylmethane: Applications and Safety of a Novel Dietary Supplement. Nutrients 2017; 9(3): 290..
29. Hari I and Mathew N: Larvicidal activity of selected plant extracts and their combination against the mosquito vectors Culexquinque fasciatus and Aedesaegypti. Environ Sci Pollut Res 2018; 25: 9176–9185. https://doi.org/10.1007/s11356-018-1515-3
30. Prakash J and Arora NK: Novel metabolites from Bacillus safensis and their antifungal property against Alternaria alternata. Antonie van Leeuwenhoek 2021; 114: 1245–1258. https://doi.org/10.1007/s10482-021-01598-4
31. Mani G: Extraction and identification of bioactive components in Sida cordata (Burm.f.) using gas chromatography–mass spectrometry. Journal of Food Science and Technology 2017; 54. Doi: 10.1007/s13197-017-2744-z.
32. Xiong H, Qi S, Xu Y, Miao L and Qian PY: Antibiotic and antifouling compound production by the marine-derived fungus Cladosporium sp. F14. J Hydro Environ Res 2009; 2: 264–271. https://doi.org/10.1016/j.jher.2008.12.002
33. Zeb A, Ullah F, Ayaz M, Ahmad S and Sadiq A: Demonstration of biological activities of extracts from Isodonrugosus Wall. Ex Benth: Separation and identification of bioactive phytoconstituents by GC-MS analysis in the ethyl acetate extract. BMC Complement Altern Med 2017; 17(1): 284. doi: 10.1186/s12906-017-1798-9. PMID: 28558679; PMCID: PMC5450350.
34. Chatterjee S, Karmakar A and Azmi SA: Antibacterial Activity of Long-Chain Primary Alcohols from Solena amplexicaulis Proc Zool Soc 2018, 71, 313–319. https://doi.org/10.1007/s12595-017-0208-0
35. Katz DH, Marcelletti JF, Khalil MH, Pope LE and Katz LR: Antiviral activity of 1-docosanol, an inhibitor of lipid-enveloped viruses including Herpes simplex. Proc Natl Acad Sci USA 1991; 88(23): 10825-9. doi: 10.1073/pnas.88.23.10825. PMID: 1660151; PMCID: PMC53024.
36. Oliveira AMD, Conserva LM, De Souza Ferro JN, BritoFd A, Lemos RPL and Barreto E: Antinociceptive and Anti-Inflammatory Effects of Octacosanol from the Leaves of Sabiceagrisea var. grisea in Mice. International Journal of Molecular Sciences 2012; 13(2): 1598-1611. https://doi.org/10.3390/ijms13021598
37. Rhetso T, Shubharani R and Roopa MS: Chemical constituents, antioxidant, and antimicrobial activity of Allium chinense Don.Futur J Pharm Sci 2022; 6: 102. https://doi.org/10.1186/s43094-020-00100-7
38. Hirpara DG, Gajera HP and Bhimani RD: The SRAP based molecular diversity related to antifungal and antioxidant bioactive constituents for biocontrol potentials of Trichoderma against Sclerotium rolfsii.Curr Genet 2016; 62: 619–641. https://doi.org/10.1007/s00294-016-0567-5
39. Moin S, Mahalakshmipriya A and Kuppusamy S: Chemical Composition and In-vitro Antimicrobial Activity of Barlerialupulina Essential Oil. Journal of Herbs, Spices & Medicinal Plants 2012; 18: 101-109. 10.1080/10496475.2011.653711.
40. Aina DA, Oloke JK, Awoyinka OA, Adebayo EA, Akoni OI and Agbolade JO: Comparative cytotoxic effect of metabolites from wild and mutant strains of Schizophylum commune grown in submerged liquid medium. Am J Res Comm 2013; 1: 219-40.
41. Shah MD, Yong YS and Iqbal M: Phytochemical investigation and free radical scavenging activities of essential oil, methanol extract and methanol fractions of Nephrolepis biserrata. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6: 269-77, https://innovareacademics.in/journals/index.php/ijpps/article/view/1512.
42. Alqahtani SS, Makeen HA, Menachery SJ and Moni SS: Documentation of bioactive principles of the flower from Carallum aretrospiciens (Ehrenb) and in vitro antibacterial activity – Part B, Arabian Journal of Chemistry 2020; 13(10): 7370-7377.
43. Suluvoy JK and Berlin Grace VM: Phytochemical profile and free radical nitric oxide (NO) scavenging activity of Averrhoa bilimbi fruit extract 3 Biotech 2017; 7(1): 85..
44. Fries E and Puttmann W: Analysis of the antioxidant butylated hydroxytoluene (BHT) in water by means of solid phase extraction combined with GC/MS. Water Research 2002; 36: 2319-27. 10.1016/S0043-1354(01)00453-5.
45. Roselin I, Srinivasan S, Mahalingam G and Sara C: Gas Chromatography and mass spectroscopy analysis of bioactive compounds of Adiantum latifolium International Research Journal of Pharmacy 2018; 9: 125-130. 10.7897/2230-8407.0910239.
46. Manjari MS, Karthi S, Ramkumar G, Muthusamy R, Natarajan D and Shivakumar MS: Chemical composition and larvicidal activity of plant extracts from Claus enadentata (Willd) (Rutaceae) against dengue, malaria, and filariasis vectors. Parasitol Res 2014; 113(7): 2475-81. doi: 10.1007/s00436-014-3896-7. Epub 2014 May 7. PMID: 24802866.
47. Padmini R, Uma Maheshwari V and Saravanan P: Identification of novel bioactive molecules from garlic bulbs: A special effort to determine the anticancer potential against lung cancer with targeted drugs. Saudi Journal of Biological Sciences 2020; 27(12): 3274-3289. DOI: 10.1016/j.sjbs.2020.09.041. PMID: 33304133; PMCID: PMC7715046.
48. Oluwasina OO, Ezenwosu IV and Ogidi CO: Antimicrobial potential of toothpaste formulated from extracts of Syzygium aromaticum, Dennettia tripetala and Jatrophacurcas latex against some oral pathogenic microorganisms. AMB Expr 2019; 9: 20. https://doi.org/10.1186/s13568-019-0744-2
49. Nagar N, Aswathanarayan JB and Vittal RR: Anti-quorum sensing and biofilm inhibitory activity of Apium graveolens L. oleoresin. J Food Sci Technol 2020; 57: 2414–2422. https://doi.org/10.1007/s13197-020-04275-y
50. Moodley K, Joseph K and Naidoo Y: Antioxidant, antidiabetic and hypolipidemic effects of Tulbaghia violacea (wild garlic) rhizome methanolic extract in a diabetic rat model. BMC Complement Altern Med 2015, 15, 408. https://doi.org/10.1186/s12906-015-0932-9
51. Youssef NH, Qari SH, Behiry SI, Dessoky ES, El-Hallous EI, Elshaer MM, Kordy A, Maresca V, Abdelkhalek A and Heflish AA: Antimycotoxigenic Activity of Beetroot Extracts against AlternariaalternataMycotoxins on Potato Crop. Applied Sciences 2021; 11(9): 4239. https://doi.org/10.3390/app11094239
52. Baky MH, Shawky EM, Elgindi MR and Ibrahim HA: Comparative Volatile Profiling of Ludwigiastolonifera Aerial Parts and Roots Using VSE-GC-MS/MS and Screening of Antioxidant and Metal Chelation Activities. ACS Omega 2021; 6(38): 24788-24794. DOI: 10.1021/acsomega.1c03627
53. Anyanwu GO, Anzaku D, Donwell CC, Usunobun U, Adegbegi AJ, Ofoha PC and Rauf K: Chemical composition and in-vitro antiobesity and in-vivo anti-hyperlipidemic effects of Ceratothecase samoides, Jatropha tanjorensis, Mucuna flagellipes, Pterocarpus mildbraedii and Piper guineense. Phytomedicine Plus 2021; 1(3): 100042, ISSN 2667-0313, https://doi.org/10.1016/j.phyplu.2021.100042.
54. Rizwana H: Chemical Composition, FTIR Studies and Antibacterial Activity of Passiflora edulis edulis (Fruit). Journal of Pure and Applied Microbiology 2019; 13(4): 2489. Gale Academic OneFile, link.gale.com/apps/doc/A674565479/AONE?u=anon~3a7a86d&sid=googleScholar&xid=ed1846d8. Accessed 6 Aug. 2022.
55. Al-Huqail AA, Elgaaly GA and Ibrahim MM: Identification of bioactive phytochemical from two Punica species using GC-MS and estimation of antioxidant activity of seed extracts. Saudi J Biol Sci 2018; 25(7): 1420-1428. doi: 10.1016/j.sjbs.2015.11.009. Epub 2015 Nov 26. PMID: 30505191; PMCID: PMC6252002.
56. Bravo-Santano N, Ellis JK, Calle Y, Keun HC, Behrends V and Letek M: Intracellular Staphylococcus aureus Elicits the Production of Host Very Long-Chain Saturated Fatty Acids with Antimicrobial Activity. Metabolites 2019; 9(7): 148. https://doi.org/10.3390/metabo9070148
57. Senyilmaz-Tiebe D, Pfaff DH, Virtue S, Schwarz KV, Fleming T, Altamura S, Muckenthaler MU, Okun JG, Vidal-Puig A, Nawroth P and Teleman AA: Dietary stearic acid regulates mitochondria in-vivo in humans. Nat Commun 2018; 9(1): 3129. doi: 10.1038/s41467-018-05614-6. PMID: 30087348; PMCID: PMC6081440.
58. Rahman J, Tareq A, Hossain MM, Sakib S, Islam M, Hazra M, Uddin ABMN, Hoque M, Nasrin S, Emran T, Capasso R, Reza ASMA and Simal-Gandara J: Biological Evaluation, DFT Calculations and Molecular Docking Studies on the Antidepressant and Cytotoxicity Activities of Cycaspectinata Buch.-Ham. Compounds. Pharmaceuticals 2020; 13: 232. 10.3390/ph13090232.
59. Hakkim FL, Al-Buloshi M and Achankunju J: Chemical composition and anti-proliferative effect of Oman's Ganoderma applanatum on breast cancer and cervical cancer cells, Journal of Taibah University Medical Sciences 2016; 11(2): 145-151, ISSN 1658-3612, https://doi.org/10.1016/j.jtumed.2016.01.007.
60. Agarwal H and Shanmugam VK: Anti-inflammatory activity screening of Kalanchoe pinnata methanol extract and its validation using a computational simulation approach, Informatics in Medicine Unlocked 2019; 14: 6-14., https://doi.org/10.1016/j.imu.2019.01.002.
61. Erwin E, Pusparohmana W, Sari I, Hairani R and Usman U: GC-MS profiling and DPPH radical scavenging activity of the bark of Tampoi (Baccaureama crocarpa). F1000Research 2019; 7: 1977.
62. Bhaskar A, Nithya V and Gopalakrishnan V: Phytochemical evaluation by GC-MS and antihyperglycemic activity of Mucuna pruriens on streptozotocin induced diabetes in rats. Journal of Chemical and Pharmaceutical Research 2011; 3: 689-696.
63. Khan M, Khan Y, Samina, Kaun L, Shah M and Idris R: Chemical composition and antioxidant activity of essential oil of leaves and flowers of Alternanthera sessilis red from Sabah. Journal of Applied Pharmaceutical Science 2016; 6: 157-161. 10.7324/JAPS.2016.601222.
64. Chao M: Antioxidant synergism between synthesised alkylated diphenylamine and dilaurylthiodipropionate in polyolefin base fluid. Journal of Thermal Analysis and Calorimetry 2014; 117(2): 925. Gale Academic OneFile, link.gale.com/apps/doc/A384543099/AONE?u=anon~7158b1e3&sid=googleScholar&xid=ce4d26b2. Accessed 17 Aug. 2022.
65. Juárez-Rodríguez MM, Cortes-López H, García-Contreras R, González-Pedrajo B, Díaz-Guerrero M, Martínez-Vázquez M, Rivera-Chávez JA, Soto-Hernández RM Ceratothecasesamoides, Castillo-Juárez I: Tetradecanoic acids with anti-virulence properties increase the pathogenicity of Pseudomonas aeruginosa in a murine cutaneous infection model. Front Cell Infect Microbiol 2021; 10: 597517. doi: 10.3389/fcimb.2020.597517. PMID: 33585272; PMCID: PMC7876447.
66. Ghazala B, Shameel M, Choudhary MI, Shahzad S and Leghari SM: Phycochemistry and bioactivity of Tetraspora (Volvocophyta) from Sindh. Pakistan Journal of Botany 2004; 36(3): 531-548.
67. Ololade Z: Pharmacological Potential of the Stem Extract of Melissa officinalis: Compositional Profile, Polyphenol, Ascorbic Acid Contents, Antioxidant, Anti-Inflammatory and Antimicrobial Activities 2018; 1: 43-52.
68. Kadhim MJ, Mohammed GJ and Hameed IH: In-vitro antibacterial, antifungal and phytochemical analysis of methanolic extract of fruit Cassia fistula. Orient J Chem 2016; 32(3).
69. Aparna V, Dileep K, Mandal P, Karthe P, Sadasivan C and Haridas M: Anti-Inflammatory Property of n-Hexadecanoic Acid: Structural Evidence and Kinetic Assessment. Chemical Biology &amp; Drug Design 2012; 80(3): 434-439.
70. Zayed MZ, Ahmad FB, Ho WS and Pang SL: GC-MS analysis of phytochemical constituents in leaf extracts of Neolamarckia cadamba (Rubiaceae) from Malaysia. Int J Pharm Pharm Sci 2014; 6(9): 123-127.
71. Innovare I: Phytochemical screening, gc-ms analysis and biological activitiys of Ipomoea eriocarpa leaf extracts. Int J Pharm Pharm Sci 2014; 6: 4.
72. Akwu NA, Naidoo Y, Singh M, Nundkumar N Ceratothecasesamoides, Lin J: Phytochemical screening, in-vitro evaluation of the antimicrobial, antioxidant and cytotoxicity potentials of Grewialasiocarpa E. Mey. ex Harv., South African Journal of Botany 2019; 123: 180-192
73. Jodallah N and Ali-Shtayeh MS: Antioxidant and antimicrobial activity of Mandragoraau tumnalis Bertol Extracts 2013.
74. Fernandes CP, Corrêa AL, Lobo JF, Caramel OP, De Almeida FB, Castro ES and Rocha L: Triterpene esters and biological activities from edible fruits of Manilkara subsericea (Mart.) Dubard, Sapotaceae. BioMed Research International 2013.
75. Rahman S, Ismail M, Shah MR, Iriti M and Shahid M: GC/MS analysis, free radical scavenging, anticancer and β-glucuronidase inhibitory activities of Trillium govanianum Bangladesh Journal of Pharmacology 2015; 10(3): 577-583.
76. Letaief T, Garzoli S, LaghezzaMasci V, Mejri J, Abderrabba M, Tiezzi A and Ovidi E: Chemical Composition and Biological Activities of Tunisian Ziziphus lotus Extracts: Evaluation of Drying Effect, Solvent Extraction, and Extracted Plant Parts. Plants 2021; 10(12): 2651. https://doi.org/10.3390/plants10122651
77. Paudel MR, Joshi PR, Chand K, Sah AK, Acharya S, Pant B and Pant B: Antioxidant, anticancer and antimicrobial effects of In-vitro developed protocorms of Dendrobium longicornu. Biotechnol Rep (Amst) 2020; 28: 00527. doi: 10.1016/j.btre.2020.e00527. PMID: 32983924; PMCID: PMC7494665.
78. Naine J, Devi CS, Mohanasrinivasan V and Vaishnavi B: Antimicrobial, Antioxidant and Cytotoxic Activity of Marine Streptomyces parvulus VITJS11 Crude Extract. Brazilian Archives of Biology and Technology 2015; 58: 198-207. 10.1590/S1516-8913201400173.
79. Swargiary A, Roy M, Boro H and Verma A: Phytochemical analysis, antiproliferative and apoptosis-inducing properties of Persicaria strigosa 2022 https://doi.org/10.21203/rs.3.rs-1383078/v1
80. Nonglang FP, Khale A and Bhan S: Phytochemical characterization of the ethanolic extract of Kaempferia galanga rhizome for anti-oxidant activities by HPTLC and GCMS. Futur J Pharm Sci 2022; 8(9). https://doi.org/10.1186/s43094-021-00394-1
81. Tassakka ACMAR, Sumule O, Massi MN, Sulfahri, Manggau M, Iskandar IW, Alam JF, Permana AD and Liao LM: Potential bioactive compounds as SARS-CoV-2 inhibitors from extracts of the marine red alga Halymenia durvillei (Rhodophyta) - A computational study. Arab J Chem 2021; 14(11): 103393. doi: 10.1016/j.arabjc.2021.103393. Epub 2021 Aug 23. PMID: 34909061; PMCID: PMC8381616.
82. Al-Marzoqi AH, Hadi MY and Hameed IH: Determination of metabolites products by Cassia angustifolia and evaluate antimicobial activity. Journal of Pharmacognosy and Phytotherapy 2016; 8(2): 25-48.
83. Musa AM, Ibrahim MA, Aliyu AB, Abdullahi MS, Tajuddeen N, Ibrahim H and Oyewale AO: Chemical composition and antimicrobial activity of hexane leaf extract of Anisopus mannii (Asclepiadaceae). J Intercult Ethnopharmacol 2015; 4(2): 129-33. doi: 10.5455/jice.20150106124652. Epub 2015 Jan 11. PMID: 26401399; PMCID: PMC4566775.
84. Thekkumalai M, Sivanandham V and Parthasarathy R: GC-MS analysis of bioactive compounds in Bryonopsis laciniosa Fruit Extract 2015; 10.13140/RG.2.2.20114.30401.
85. Nas F, Aissaoui N and Mahjoubi M: A comparative GC–MS analysis of bioactive secondary metabolites produced by halotolerant Bacillus spp. isolated from the Great Sebkha of Oran. Int Microbiol 2021; 24: 455–470. https://doi.org/10.1007/s10123-021-00185-x
86. Abbassy MMS, Salem MZM and Rashad NM: Nutritive and biocidal properties of agroforestry trees of Moringa oleifera , Cassia fistula L., and Ceratonia siliqua L. as non-conventional edible vegetable oils. Agroforest Syst 2020; 94: 1567–1579.
87. Baskaran XR, Geo Vigila AV, Zhang SZ, Feng SX and Liao WB: A review of the use of pteridophytes for treating human ailments. Journal of Zhejiang University-Science B 2018; 19(2): 85-119.
88. Sureshkumar J, Silambarasan R, Bharati KA, Krupa J, Amalraj S & Ayyanar M: A review on ethnomedicinally important pteridophytes of India. Journal of Ethnopharmacology 2018; 219: 269-287.
    

    ---

    How to cite this article:

    Borkotoky S and Borah VV: Antimicrobial potential of crude extracts and fatty acid fractions of four pteridophyte species from Assam, Northeast India, and their identification by GC-MS. Int J Pharm Sci & Res 2023; 14(9): 4489-08. doi: 10.13040/IJPSR.0975-8232.14(9).4489-08.

    ---

    All © 2023 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.