Next Article in Journal
Study on Separation of Rhenium, a Surrogate Element of Fissiogenic Technetium, from Aqueous Matrices Using Ion-Selective Extraction Chromatographic Resins
Next Article in Special Issue
Box–Behenken-Supported Development and Validation of UPLC Method for the Estimation of Eugenol in Syzygium aromaticum, Cinnamomum tamala, and Myristica fragrance
Previous Article in Journal
Mechanism of Separation of Contaminants from Activated Carbon by Closed Cycle Temperature Swing Desorption
Previous Article in Special Issue
The Role of Selective Flavonoids on Triple-Negative Breast Cancer: An Update
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:

LC-MS/MS and GC-MS Analysis for the Identification of Bioactive Metabolites Responsible for the Antioxidant and Antibacterial Activities of Lygodium microphyllum (Cav.) R. Br.

Faculty of Pharmacy and Health Sciences, Universiti Kuala Lumpur-Royal College of Medicine Perak, Ipoh 30450, Malaysia
Laboratory Centre, Xiamen University Malaysia, Jalan Sunsuria, Bandar Sunsuria, Sepang 43900, Malaysia
School of Applied Sciences, Faculty of Integrated Life Sciences, Quest International University, Jalan Raja Permaisuri Bainun, Ipoh 30250, Malaysia
Borneo Marine Research Institute, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu 88400, Malaysia
Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur 56000, Malaysia
Department of Nutrition, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Malaysia
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Separations 2023, 10(3), 215;
Received: 23 February 2023 / Revised: 12 March 2023 / Accepted: 15 March 2023 / Published: 20 March 2023


Natural products serve as a valuable source of antioxidants with potential health benefits for various conditions. Lygodium microphyllum (Cav.) R. Br., also known as Old World climbing fern, is an invasive climbing fern native to Southeast Asia, Africa, South America, Australia, and Melanesia. It has been reported to possess interesting pharmacological properties including hepatoprotective and anti-inflammatory mechanisms. This study analyzed the potential bioactive metabolites that contribute to the antioxidant and antimicrobial effects of L. microphyllum (LM) by profiling the crude extract using high-resolution LC-MS/MS and GC-MS systems. Several classes of compounds such as phenolics, flavonoids, terpenoids, steroids, macrolides, vitamins, lipids, and other hydrocarbons were found in the crude extract of LM through non-targeted analysis. A total of 74 compounds were detected in LC-MS/MS, whereas a total of nine compounds were identified in GC-MS. Out of the 74 compounds detected in LC-MS/MS, 34 compounds, primarily quercetin, kaempferol, trifolin, pyroglutamic acid, arachidonic acid, and rutin were reported with antioxidant, antimicrobial, anti-inflammatory, and hepatoprotective activities. The presence of phenolic and flavonoid compounds with reported bioactivities in the crude extract of LM evidence its pharmacological properties.

1. Introduction

Medicinal plants have many important roles in the pharmaceutical and food industries. The phytochemical compounds obtained from plants have been developed into commercial medicine and have always been a source for the discovery of new medicinal drugs. Bioactive compounds of plant origin are known to contribute to human health improvement, especially in treating diseases related to oxidative stress. Flavonoids are the largest group of polyphenolic compounds that are present in high concentrations in medicinal plants [1]. These compounds majorly contribute to the proclaimed pharmacological properties of the plants and have also been widely reported to possess therapeutic effects as individual compounds. Numerous polyphenols have been studied for their therapeutic values, and most of the compounds have been developed into commercial drugs for various diseases [2]. The ability to scavenge free radicals is the fundamental requirement for a bioactive compound to exert pharmacological effects.
Free radicals are formed during the body’s normal physiological activities but can be controlled under normal conditions due to the presence of an antioxidant defense system comprising reduced glutathione and antioxidant enzymes [3]. Excessive free radical formation is caused by unfavorable conditions such as ingestion of toxicants or exposure to radiation, thus inflicting an imbalance in the body’s antioxidant defense system, which leads to oxidative stress. Numerous free radicals in the body at the cellular level cause an increase in malondialdehyde (MDA) formation, which is also a result of lipid peroxidation at the cell membrane. This formation triggers a cascade of events that could lead to tissue damage and eventually organ failures. Lipid peroxidation at the cellular level implies the activation of inflammatory and apoptosis events [4]. Oxidative stress and inflammation remain pathophysiological factors for the development of many ailments, including cancer, liver damage, diabetes, brain disorders, and heart problems [5]. Therefore, bioactive compounds from plants are among the most suitable drug candidates to counter oxidative stress-related diseases by exerting antioxidant effects. External supplementation of antioxidants has been proven to accelerate the endogenous antioxidant defense mechanism as these antioxidant compounds have the tendency to pass the gastrointestinal barrier as cytochrome P-450 enzymes cannot alter the compounds and cause them to be excreted out of the body. Bioactive compounds that are detected as xenobiotics in the liver could alter the mechanism of cytochrome P-450 by escaping biotransformation and hence could be present as an active compound in the same chemical configuration to reach the target site [6]. Crude extracts of medicinal plants tend to express synergistic effects in regard to antioxidant activities with minimal or no toxicity, especially in preclinical assessments. Bioactive compounds, especially flavonoids, tend to be commercialized into therapeutic drugs, but the bioavailability of the individual compounds in in vivo trials remains a hurdle. Therefore, the administration of crude extracts or a mixture of bioactive compounds could be regarded as a suitable measure to overcome the problem of bioavailability.
Lygodium microphyllum (Cav.) R. Br. (LM) is an invasive fern species native to American and Asian countries. Although this invasive plant is considered a threat to the forest as it grows rapidly, it contains important phytochemical compounds that are useful for the treatment of diseases. The native community of Sabah claimed that this plant could be used for the treatment of skin ailments and dysentery through the consumption of aqueous decoctions. Our previous study on LM reported the hepatoprotective and immunosuppressive effects and effective antioxidant properties in the aqueous extracts of the leaves [7]. The antioxidant properties of LM in an aqueous extract were noteworthy, with the total phenolic content of 206.38 ± 9.62 mg/g gallic acid equivalent (GAE) and the IC50 value for the 2,2-diphenylpicrylhydrazine (DPPH) study of 65 µg/mL. Other reports on the anthelmintic, antipyretic, anti-inflammatory, and antioxidant activities and the qualitative phytochemical analysis of LM support the medicinal claims made about this plant [8]. This study was conducted to screen the potential bioactive compounds present in an aqueous extract of LM using liquid chromatography–high resolution tandem mass spectrometry (LC-MS/MS) and gas chromatography–mass spectrometry (GC-MS) and to evaluate its antibacterial properties on several bacterial strains.

2. Materials and Methods

2.1. Plant Collection and Extraction

The fresh plant specimen was collected from the area of Papar, Kota Kinabalu, Sabah, and a voucher specimen was deposited at Universiti Malaysia Sabah (voucher number CG005). The plant specimen was verified by a field botanist of the same institute upon collection from the wild. The mature leaves (1 kg) were cleaned, air-dried for 5 days, homogenized using a heavy-duty blender, and subjected to aqueous extraction as described previously in a ratio of 1:10 in distilled water [6]. Briefly, 100 g of LM was boiled in 1 L of distilled water for 10 min, followed by cooling for 1 h and filtration. The extracts were lyophilized and kept at −20 °C for further analysis.

2.2. LC-MS/MS Analysis

High-resolution MS/MS analysis was performed as described by Haron et al. [8] using the Thermo Scientific Q Exactive HF Orbitrap mass spectrometry system (Thermo Fisher Scientific, Waltham, MA, USA). Prior to the analysis, metabolite separation was performed with the Dionex UltiMate 3000 ultra-high-performance liquid chromatography system (Thermo Fisher Scientific, Waltham, MA, USA) coupled with a Thermo Syncronis C18 column (2.1 mm × 100 mm × 1.7 µm; Thermo Fisher Scientific, Waltham, MA, USA). The column was maintained at 55 °C at a flow rate of 450 µL/min during analysis. All instrumental settings, elution gradients, and calibration were performed as described previously [9,10]. The mobile phases were prepared with HPLC-grade deionized water with formic acid 0.1% (Solvent A) and acetonitrile with formic acid 0.1% (Solvent B). The elution gradient program was started with 0.5% of Solvent B for 1 min, followed by 0.5% gradually to 95.5% of Solvent B for 15 min which was maintained for 4 min. The injection volume of the sample was set at 2 µL. The column was later conditioned for the next injection through flushing for 2 min as the initial cycle.
The acquired data were processed and analyzed using the Thermo Scientific Compound Discoverer 3.3 SP1 software (Thermo Fisher Scientific, Waltham, MA, USA) with minor adjustments on the default settings for the natural product workflow. Briefly, the workflow included background subtraction with blank data, retention time alignment, feature detection, elemental composition determination, library matching, and fragment ion search (FISh) scoring. The identification of compounds was primarily based on the matching of MS/MS data against the mzCloud database. Identification of unmatched signals was re-attempted on the ChemSpider database [11] using MS data and supported with a FISh scoring of above 50.

2.3. GC-MS Analysis

LM aqueous extract was diluted in methanol, and the sample was injected (1 µL) into a GC-MS system consisting of a gas chromatography system (Agilent 7890A) coupled with a mass spectrometry detector (Agilent 5975C). An HP-5MS (30 m × 0.25 mm) capillary column was used with 0.25 µm film thickness of coated material. The injector temperature (250 °C) was set; the temperature program was as follows: starting at 40 °C, hold for 3 min, from 40 to 300 °C (3 °C/min), and hold for 3 min. A post-run at 300 °C for 5 min was performed to prepare for the next injection. Gas chromatography was performed in spitless mode using helium gas as a carrier at a constant flow rate of 1 mL/min. Compounds were identified with reference to the NIST 11 library, and compositions were computed with reference to the abundance of compounds in the chromatogram. The complete analysis was performed in triplicate, together with a blank solvent.

2.4. Disc Diffusion Assay for Antibacterial Activity

Staphylococcus aureus (ATCC 33862), Bacillus cereus (ATCC 14579), Escherichia coli (ATCC 25922), and Salmonella sp. (ATCC 29890) were used as test bacteria to assess the antibacterial activity of LM. All test bacteria were obtained from UniKL-RCMP and cultured using Mueller–Hinton agar (MHA). The antibacterial activity of LM was evaluated using the disc diffusion method [12]. First, test bacteria were suspended in 0.9% saline solution. The optical density (OD) of bacterial suspension was adjusted to match the 0.5 McFarland standard (0.08 to 0.12 at 625 nm). Then, 100 μL of the bacterial suspension was pipetted onto the MHA plate to prepare bacterial lawn using the spread plate technique. LM aqueous extract was dissolved in 0.9% saline solution into two different concentrations (500 mg/mL and 100 mg/mL) to test for antibacterial activity. Then, 20 µL of the sample were pipetted onto sterile filter paper discs (Whatman No. 3; 6 mm diameter) and placed on the surface of the agar. A 10 µg gentamicin antibiotic disc (Oxoid) was used as a positive control, while a filter paper disc with 0.9% saline solution added acted as a negative control. The plates were incubated at 37 °C for 18 h. The diameter of the inhibition zone (mm) was measured, and the experiment was performed in triplicates.

2.5. Determination of Minimum Inhibitory Concentration (MIC)

The broth microdilution method was used to determine MIC [12]. LM water extract was dissolved in Mueller–Hinton broth to achieve 12.5 mg/mL. One hundred microliters of 12.5 mg/mL water extract was loaded into each well of a 96-well plate and 2-fold serially diluted using Mueller–Hinton broth (50 µL in each well as diluent). Then, Staphylococcus aureus suspension was prepared with an OD of 0.08 to 0.12 at 625 nm wavelength. This bacterial suspension was diluted 150 times using Mueller–Hinton broth to obtain an inoculum at 106 CFU/mL. Fifty microliters of inoculum was added into each well containing the extract. The 96-well plate was incubated for 18 h at 37 °C. The MIC was determined based on the lowest water extract concentration with the absence of turbidity. Gentamicin was used as positive control, whereas Mueller–Hinton broth was used as negative control.

2.6. Statistical Analysis

All data were analyzed using SPSS 25.0 (IBM, Armonk, NY, USA) Windows statistical package. Results are shown as mean ± standard deviation of triplicate measurements.

3. Results and Discussion

3.1. LM Extraction Yield and Antioxidant Properties

The extraction yield of LM was 18.3% for aqueous extract, which is considered sufficient for water extracts. In our previous study, we performed the extraction of LM with different solvents in various polarities, i.e., methanol, hexane, ethyl acetate, chloroform, and butanol [13]. Total phenolic content (TPC), total flavonoid content (TFC), and DPPH antioxidant tests were performed for all the extracts. The TPC was high in hexane (354.38 ± 0.57 mg/g GAE) and ethyl acetate (347.18 ± 0.28 mg/g GAE) extracts but was also comparable in the other extracts. The aqueous extract of LM had a TFC of 20.68 ± 3.67 mg/g catechin equivalent (CE), which was slightly lower than the TFC of the methanol extract (39.36 ± 2.73 mg/g CE), the highest TFC among the extracts. The DPPH results were convincing, with effective antioxidant effects shown in all the extracts of LM. IC50 values for the solvent extracts, i.e., methanol (60 µg/mL), ethyl acetate (52 µg/mL), hexane (61 µg/mL), butanol (81 µg/mL), and chloroform (76 µg/mL), were also comparable with that of the aqueous extract (65 µg/mL). The reason for choosing aqueous extract for this study is to validate the medicinal claim made by the indigenous people of Sabah that an aqueous decoction of the leaves of LM can treat several ailments. Moreover, we had previously published results showing the hepatoprotective effect of LM aqueous extract against carbon tetrachloride (CCl4)-induced liver damage in rats [6]. The metabolites responsible for hepatoprotective and immunosuppressive effects were not elucidated in our previous study. These results indicate that the aqueous extract of LM had adequate phenolic and flavonoid contents although organic solvents had slightly higher TPC and TFC contents. The DPPH results indicate the ability of the aqueous extract to demonstrate excellent antioxidant effects by scavenging free radicals. This property is essential for therapeutic interventions, especially for oxidative stress-based diseases. It is suggested that organic solvent extracts of LM such as hexane, ethyl acetate, and methanol with higher TPC and TFC values should be evaluated for their pharmacological properties and the bioactive compounds responsible for the activities should be elucidated.

3.2. Identified Bioactive Compounds in LC-MS/MS

A total of 74 compounds were identified (59 in positive mode and 15 in negative mode) in the aqueous extract of LM using the MS/MS spectra (Table 1 and Table 2). A few important classes of bioactive compounds such as flavonoids, phenolics, terpenoids, steroids, alkaloids, and vitamin B were detected in the extract of LM. Out of the 74 compounds, 13 flavonoids, 12 phenolic, and 6 terpenoids were detected as major contributors to the antioxidant activity and pharmacological functions of LM. The remaining classes of compounds such as fatty acids, aromatic compounds, amino acids, lactones, and heterocyclic ketones also partly contributed to the medicinal value of LM since some of the compounds were reported to possess pharmacological properties [7].
Our previous study on the hepatoprotective and immunosuppressive effects of LM aqueous extract reported the ability of LM to reverse the effect of CCl4 administration in rats and demonstrated the immunosuppressive effect on proinflammatory cytokines and oxidative stress markers [6]. The bioactive compounds identified in LM aqueous extract that could potentially be responsible for the hepatoprotective and anti-inflammatory effects are jasmonic acid [14], maltol [15,16], kaempferol [17,18], luteolin 7-O-malonylglucoside [19], nicotiflorin [20,21], quercetin [22,23], robinin [24], trifolin [25], adenine [26], adenosine [27,28], coniine [29], guanine [30], massoilactone [31], caffeic acid [32,33], coniferol [34], demethoxycurcumin [35], esculin [36,37], paradol [38], shogaol [39,40], (3R)-hydroxy-beta-ionone [41], caryophyllene oxide [42,43], costunolide [44,45], nootkatone [46,47], nicotinamide [48,49], pantothenic acid [50], pyridoxine [51,52], astragalin [53,54], and rutin [55,56].
Two unknown steroidal compounds were also detected in the aqueous extract of LM but were not identified since the identification of underivatized steroidal compounds via tandem mass spectrometry is impossible and any putative identity could be misleading due to their stable four-ring skeleton and diverse stereoisomerisms [57]. Kuncoro et al. [58] reported two new steroidal compounds in a methanol extract of LM, stigmast-5 (6)-en-3β-ol and stigmast-4-en-3-one, identified using NMR spectroscopy. Hence, the two unidentified steroidal compounds detected in MS/MS analysis could be the same as those reported earlier or their derivatives, as the suggested molecular formulas of the compounds are almost the same.

3.3. Identification of Metabolites in GC-MS

The aqueous extract of LM was subjected to GC-MS analysis to screen volatile bioactive compounds that might also be responsible for the pharmacological properties of this plant. The compounds identified are listed in Table 3. The identified bioactive compounds from different classes such as cyclic aldehyde (5-hydroxymethylfurfural), phenolic acid (p-coumaric acid), and fatty acyls (E-15-heptadecenal, n-hexadecanoic acid) were reported to possess pharmacological properties such as anti-inflammatory, anticancer, and antimicrobial effects [59,60,61,62]. The detected volatile compounds could have exerted synergistic effects to mitigate oxidative stress by enhancing the antioxidant and immunosuppressive properties of LM. GC-MS analysis was performed to identify the volatile compounds that could have been missed in the LC-MS/MS analysis. LM leaves exerted a strong aroma while being boiled for extraction, indicating the presence of essential oils and other volatile aromatic compounds in the leaves. Hence, the leaves of LM can be subjected to essential oil extraction and organic solvent extracts for a future direction to elucidate the volatile compounds present using GC-MS analysis since most of the thermolabile compounds would be lost due to high temperature during boiling for aqueous extract.

3.4. Antibacterial Effect of LM

LM was tested against two Gram-positive bacteria (S. aureus and B. cereus) and two Gram-negative bacteria (E. coli and Salmonella sp.). LM showed antibacterial activity at high and low concentrations against S. aureus with 15 mm and 13 mm inhibition zones, respectively, but did not show activity against B. cereus (Table 4). This suggested certain Gram-positive bacteria species are susceptible to the phytochemicals present in the aqueous extract. LM was not able to inhibit the growth of the tested Gram-negative bacteria. There is a distinct difference between Gram-positive and Gram-negative bacterial cell walls. Gram-negative bacteria have three cell wall layers (outer membrane, peptidoglycan, and inner membrane), while Gram-positive bacteria have two cell wall layers (peptidoglycan and inner membrane) only. The extra outer membrane of Gram-negative bacteria is the main reason for their lower sensitivity to many antibacterial compounds [63]. The antibacterial effect of LM aqueous extract was further determined to estimate the MIC against S. aureus using the broth microdilution method. The results indicated LM required more than 6.25 mg/mL to achieve the MIC value. This value is considered low as compared to the positive control gentamicin that exhibited MIC at 2.5 µg/mL. The broth microdilution method used for the determination of MIC in LM aqueous extract showed the cloudy and intense color of the plant extract, which interfered with the MIC evaluation. Therefore, a disc diffusion test on the aqueous extract of LM at low concentration should be performed to determine the exact MIC value. However, the MIC value of LM was not strong enough to motivate the continuation of the experiment. Compounds with antioxidant properties are known to possess antimicrobial effects [64,65]. The bioactive compounds that could have contributed to the antibacterial activities of LM, apart from flavonoids, phenolics, and terpenoids, are fatty acyl groups and amino acids such as pyroglutamic acid [66], 3-indoleacrylic acid [67], trifolin [68], and arachidonic acid [69].
The negative control did not show a growth inhibition effect. Values are given as mean ± SD of three replicate samples.

4. Conclusions

Screening of bioactive compounds in the aqueous extract of LM using LC-MS/MS and GC-MS resulted in the successful identification of flavonoids, phenolics, terpenoids, amino acids, cyclic ketones, lactones, amino acids, fatty acyls, and aromatic compounds that contributed to the antioxidant, hepatoprotective, anti-inflammatory, and antibacterial activities of the plant. Almost half of the list of compounds identified were reported to have pharmacological properties. The high presence of flavonoids such as quercetin, kaempferol, trifolin, and rutin could be attributed to the efficient antioxidant, free radical scavenging, and antibacterial effects of LM. Therefore, the presence of important phytochemicals could be responsible for the medicinal properties of LM against diseases prevailing from oxidative stress. It can be suggested that organic solvent extracts of LM such as methanol, ethyl acetate, and hexane could potentially have additional bioactive compounds; therefore, studies should be directed to identify the metabolites in the extracts as well to evaluate their pharmacological properties on various diseases relating to oxidative stress.

Author Contributions

W.Y.T. and Y.S.Y. equally contributed to the curation and writing the manuscript; F.N.R., S.S. and J.K.T. contributed to data analysis; M.D.S. and N.M.E. contributed in editing the manuscript; C.G. contributed in conceptualization and writing the manuscript. All authors have read and agreed to the published version of the manuscript.


This research was funded by the Ministry of Higher Education (MOHE), Malaysia via the Fundamental Research Grant Scheme [Ref: FRGS/1/2020/SKK06/UNIKL/03/2].

Data Availability Statement

The data presented in this study are available on request from the corresponding author.


The authors Charles Gnanaraj, Norhaizan Mohd Esa, and Sarah Stephenie would like to thank the Ministry of Higher Education (MOHE), Malaysia, for the financial support provided via the Fundamental Research Grant Scheme (FRGS/1/2020/SKK06/UNIKL/03/2).

Conflicts of Interest

The authors declare no conflict of interest.


  1. Tang, J.; Dunshea, F.R.; Suleria, H.A. LC-ESI-QTOF/MS characterization of phenolic compounds from medicinal plants (hops and juniper berries) and their antioxidant activity. Foods 2019, 9, 7. [Google Scholar] [CrossRef] [PubMed][Green Version]
  2. Chen, L.; Gnanaraj, C.; Arulselvan, P.; El-Seedi, H.; Teng, H. A review on advanced microencapsulation technology to enhance bioavailability of phenolic compounds: Based on its activity in the treatment of type 2 diabetes. Trends Food Sci. Technol. 2019, 85, 149–162. [Google Scholar] [CrossRef]
  3. Alsarhan, A.; Sultana, M.; Khatib, A.A.; Kadir, M.R.A. Review on some Malaysian traditional medicinal plants with therapeutic properties. J. Basic Appl. Sci. 2014, 10, 149–159. [Google Scholar] [CrossRef][Green Version]
  4. Amoateng, P.; Koffuor, G.A.; Sarpong, K.; Agyapong, K.O. Free radical scavenging and anti-lipid peroxidative effects of a hydro-ethanolic extract of the whole plant of Synedrella nodiflora (L.) Gaertn (asteraceae). Free Rad. Antiox. 2011, 1, 70–78. [Google Scholar] [CrossRef][Green Version]
  5. Gnanaraj, C.; Shah, M.D.; Makki, J.S.; Iqbal, M. Hepatoprotective and immunosuppressive effect of Synedrella nodiflora L. in carbon tetrachloride (CCl4)-intoxicated rats. J. Environ. Pathol. Toxicol. Oncol. 2016, 35, 29–42. [Google Scholar] [CrossRef] [PubMed][Green Version]
  6. Androutsopoulos, V.P.; Papakyriakou, A.; Vourloumis, D.; Tsatsakis, A.M.; Spandidos, D.A. Dietary flavonoids in cancer therapy and prevention: Substrates and inhibitors of cytochrome P450 CYP1 enzymes. Pharmacol. Ther. 2010, 126, 9–20. [Google Scholar] [CrossRef]
  7. Gnanaraj, C.; Shah, M.D.; Song, T.T.; Iqbal, M. Hepatoprotective mechanism of Lygodium microphyllum (Cav.) R.Br. through ultrastructural signaling prevention against carbon tetrachloride (CCl4)-mediated oxidative stress. Biomed. Pharmacother. 2017, 92, 1010–1022. [Google Scholar] [CrossRef] [PubMed]
  8. Alam, M.M.; Emon, N.U.; Alam, S.; Rudra, S.; Akhter, N.; Mamun, M.M.; Ganguly, A. Assessment of pharmacological activities of Lygodium Microphyllum Cav. leaves in the management of pain, inflammation, pyrexia, diarrhea, and helminths: In vivo, in vitro and in silico approaches. Biomed. Pharmacother. 2021, 139, 111644. [Google Scholar] [CrossRef]
  9. Haron, F.K.; Shah, M.D.; Yong, Y.S.; Tan, J.K.; Lal, M.T.M.; Venmathi Maran, B.A. Antiparasitic potential of methanol extract of brown alga Sargassum polycystum (Phaeophyceae) and its LC-MS/MS metabolite profiling. Diversity 2022, 14, 796. [Google Scholar] [CrossRef]
  10. Shah, M.D.; Venmathi Maran, B.A.; Tan, J.K.; Yong, Y.S.; Fui Fui, C.; Shaleh, S.R.M.; Shapawi, R. The anti-leech potential of the solvent extract of Bornean neem leaves and ultra-high performance liquid chromatography-high-resolution mass spectrometry profiling. J. King Saud Univ. Sci. 2021, 33, 101541. [Google Scholar] [CrossRef]
  11. Venmathi Maran, B.A.; Josmeh, D.; Tan, J.K.; Yong, Y.S.; Shah, M.D. Efficacy of the aqueous extract of Azadirachta indica against the marine parasitic leech and its phytochemical profiling. Molecules 2021, 26, 1908. [Google Scholar] [CrossRef]
  12. Pence, H.E.; Williams, A. ChemSpider: An Online Chemical Information Resource. J. Chem. Educ. 2021, 87, 1123–1124. [Google Scholar] [CrossRef]
  13. Balouiri, M.; Sadiki, M.; Ibnsouda, S.K. Methods for in vitro evaluating antimicrobial activity: A review. J. Pharm. Anal. 2016, 6, 71–79. [Google Scholar] [CrossRef] [PubMed][Green Version]
  14. Gnanaraj, C.; Iqbal, M. Total phenolic contents and free radical scavenging activity in various extracts of Lygodium microphyllum. Short Communic. Biotech. 2017, 4, 35–42. [Google Scholar]
  15. Jarocka-Karpowicz, I.; Markowska, A. Therapeutic potential of jasmonic acid and its derivatives. Int. J. Mol. Sci. 2021, 22, 8437. [Google Scholar] [CrossRef]
  16. Zhu, D.; Wang, Y.; Lin, J.; Miao, Z.; Xu, J.; Wu, Y. Maltol inhibits the progression of osteoarthritis via the nuclear factor-erythroid 2–related factor-2/heme oxygenase-1 signal pathway in vitro and in vivo. Food Func. 2021, 12, 1327–1337. [Google Scholar] [CrossRef]
  17. Han, Y.; Xu, Q.; Hu, J.; Han, X.; Li, W.; Zhao, L. Maltol, a food flavoring agent, attenuates acute alcohol-induced oxidative damage in mice. Nutrients 2015, 7, 682–696. [Google Scholar] [CrossRef][Green Version]
  18. Zang, Y.; Zhang, D.; Yu, C.; Jin, C.; Igarashi, K. Antioxidant and hepatoprotective activity of kaempferol 3-O-β-D-(2,6-di-O-α-L-rhamnopyranosyl)galactopyronoside against carbon tetrachloride-induced liver injury in mice. Food Sci. Biotechnol. 2017, 26, 1071–1076. [Google Scholar] [CrossRef] [PubMed]
  19. Alam, W.; Khan, H.; Shah, M.A.; Cauli, O.; Saso, L. Kaempferol as a dietary anti-inflammatory agent: Current therapeutic standing. Molecules 2020, 25, 4073. [Google Scholar] [CrossRef] [PubMed]
  20. Park, C.M.; Song, Y. Luteolin and luteolin-7-o-glucoside protect against acute liver injury through regulation of inflammatory mediators and antioxidative enzymes in Galn/LPS-induced hepatitic ICR MICE. Nutr. Res. Pract. 2019, 13, 473–479. [Google Scholar] [CrossRef] [PubMed]
  21. Yu, S.; Guo, Q.; Jia, T.; Zhang, X.; Guo, D.; Jia, Y.; Li, J.; Sun, J. Mechanism of action of nicotiflorin from Tricyrtis maculata in the treatment of acute myocardial infarction: From Network Pharmacology to Experimental Pharmacology. Drug Des. Devel. Ther. 2021, 15, 2179–2191. [Google Scholar] [CrossRef] [PubMed]
  22. Zhao, J.; Zhang, S.; You, S.; Liu, T.; Xu, F.; Ji, T.; Gu, Z. Hepatoprotective effects of Nicotiflorin from Nymphaea candida against concanavalin a-induced and D-galactosamine-induced liver injury in mice. Int. J. Mol. Sci. 2017, 8, 587. [Google Scholar] [CrossRef] [PubMed][Green Version]
  23. Li, Y.; Yao, J.; Han, C.; Yang, J.; Chaudhry, M.; Wang, S.; Liu, H.; Yin, Y. Quercetin, inflammation and immunity. Nutrients 2016, 8, 167. [Google Scholar] [CrossRef] [PubMed][Green Version]
  24. Miltonprabu, S.; Tomczyk, M.; Skalicka-Woźniak, K.; Rastrelli, L.; Daglia, M.; Nabavi, S.F.; Alavian, S.M.; Nabavi, S.M. Hepatoprotective effect of quercetin: From chemistry to medicine. Food Chem. Toxicol. 2017, 108, 365–374. [Google Scholar] [CrossRef] [PubMed]
  25. Tsiklauri, L.; Švík, K.; Chrastina, M.; Poništ, S.; Dráfi, F.; Slovák, L.; Alania, M.; Kemertelidze, E.; Bauerova, K. Bioflavonoid Robinin from Astragalus falcatus lam. mildly improves the effect of Metothrexate in rats with adjuvant arthritis. Nutrients 2021, 13, 1268. [Google Scholar] [CrossRef]
  26. Kim, S.M.; Kang, K.; Jho, E.H.; Jung, Y.; Nho, C.W.; Um, B.; Pan, C. Hepatoprotective effect of flavonoid glycosides from Lespedeza cuneata against oxidative stress induced by tert-butyl hyperoxide. Phytother. Res. 2011, 25, 1011–1017. [Google Scholar] [CrossRef]
  27. Chen, Y.; Chu, Y.; Tsuang, Y.; Wu, Y.; Kuo, C.; Kuo, Y. Anti-inflammatory effects of adenine enhance osteogenesis in the osteoblast-like MG-63 cells. Life 2020, 10, 116. [Google Scholar] [CrossRef]
  28. Tanaka, Y.; Ohashi, S.; Ohtsuki, A.; Kiyono, T.; Park, E.Y.; Nakamura, Y.; Sato, K.; Oishi, M.; Miki, H.; Tokuhara, K.; et al. Adenosine, a Hepato-protective component in active hexose correlated compound: Its identification and inos suppression mechanism. Nitric Oxide 2014, 40, 75–86. [Google Scholar] [CrossRef] [PubMed]
  29. Haskó, G.; Cronstein, B. Regulation of inflammation by adenosine. Front. Immunol. 2013, 4, 75–86. [Google Scholar] [CrossRef] [PubMed][Green Version]
  30. Arihan, O.; Boz, M.; Iskit, A.B.; Ilhan, M. Antinociceptive activity of coniine in mice. J. Ethnopharm. 2009, 125, 274–278. [Google Scholar] [CrossRef]
  31. Tasca, C.I.; Lanznaster, D.; Oliveira, K.A.; Fernández-Dueñas, V.; Ciruela, F. Neuromodulatory effects of guanine-based purines in health and disease. Front. Cell. Neurosci. 2018, 12, 376. [Google Scholar] [CrossRef] [PubMed][Green Version]
  32. Barros, M.E.; Freitas, J.C.; Oliveira, J.M.; Da Cruz, C.H.; Da Silva, P.B.; De Araújo, L.C.; Militão, G.C.; da Silva, T.G.; Oliveira, R.A.; Menezes, P.H. Synthesis and evaluation of (−)-massoialactone and analogues as potential anticancer and anti-inflammatory agents. Eur. J. Med. Chem. 2014, 76, 291–300. [Google Scholar] [CrossRef]
  33. Choi, H.G.; Tran, P.T.; Lee, J.; Min, B.S.; Kim, J.A. Anti-inflammatory activity of caffeic acid derivatives isolated from the roots of Salvia Miltiorrhiza Bunge. Arch. Pharm. Res. 2018, 41, 64–70. [Google Scholar] [CrossRef] [PubMed]
  34. Yang, S.; Hong, C.; Lee, G.P.; Kim, C.; Lee, K. The hepatoprotection of caffeic acid and rosmarinic acid, major compounds of perilla frutescens, against T-bhp-induced oxidative liver damage. Food Chem. Toxicol. 2013, 55, 92–99. [Google Scholar] [CrossRef]
  35. Wang, Y.; Gao, Y.; Li, X.; Sun, X.; Wang, Z.; Wang, H.; Nie, R.; Yu, W.; Zhou, Y. Coniferyl aldehyde inhibits the inflammatory effects of leptomeningeal cells by suppressing the JAK2 signaling. BioMed Res. Int. 2020, 2020, 4616308. [Google Scholar] [CrossRef] [PubMed]
  36. Hatamipour, M.; Ramezani, M.; Tabassi, S.A.; Johnston, T.P.; Sahebkar, A. Demethoxycurcumin: A naturally occurring curcumin analogue for treating non-cancerous diseases. J. Cell. Physiol. 2019, 234, 19320–19330. [Google Scholar] [CrossRef]
  37. Yang, X.; Chen, Z.; Ye, L.; Chen, J.; Jang, Y. Esculin protects against methionine choline-deficient diet-induced non-alcoholic steatohepatitis by regulating the SIRT1/NF-ΚB P65 pathway. Pharm. Biol. 2021, 59, 920–930. [Google Scholar] [CrossRef]
  38. Niu, X.; Wang, Y.; Li, W.; Zhang, H.; Wang, X.; Mu, Q.; He, Z.; Yao, H. Esculin exhibited anti-inflammatory activities in vivo and regulated TNF-α and IL-6 production in LPS-stimulated mouse peritoneal macrophages in vitro through MAPK pathway. Int. Immunopharmacol. 2015, 29, 779–786. [Google Scholar] [CrossRef]
  39. Rafeeq, M.; Murad, H.A.; Abdallah, H.M.; El-Halawany, A.M. Protective effect of 6-paradol in acetic acid-induced ulcerative colitis in rats. BMC Complement. Med. Ther. 2021, 21, 60. [Google Scholar] [CrossRef]
  40. Zhang, H.; Wang, Q.; Sun, C.; Zhu, Y.; Yang, Q.; Wei, Q.; Chen, J.; Deng, W.; Adu-Frimpong, M.; Yu, J.; et al. Enhanced oral bioavailability, anti-tumor activity and hepatoprotective effect of 6-shogaol loaded in a type of novel micelles of polyethylene glycol and linoleic acid conjugate. Pharmaceutics 2019, 11, 107. [Google Scholar] [CrossRef][Green Version]
  41. Shim, S.; Kim, S.; Choi, D.; Kwon, Y.; Kwon, J. Anti-inflammatory effects of [6]-shogaol: Potential roles of HDAC inhibition and hsp70 induction. Food Chem. Toxicol. 2011, 49, 2734–2740. [Google Scholar] [CrossRef] [PubMed]
  42. Aloum, L.; Alefishat, E.; Adem, A.; Petroianu, G. Ionone is more than a Violet’s fragrance: A Review. Molecules 2020, 25, 5822. [Google Scholar] [CrossRef] [PubMed]
  43. Chavan, M.; Wakte, P.; Shinde, D. Analgesic and anti-inflammatory activity of caryophyllene oxide from Annona squamosa L. Bark. Phytomedicine 2010, 17, 149–151. [Google Scholar] [CrossRef]
  44. Francomano, F.; Caruso, A.; Barbarossa, A.; Fazio, A.; La Torre, C.; Ceramella, J.; Mallamaci, R.; Saturnino, C.; Iacopetta, D.; Sinicropi, M.S. B-caryophyllene: A sesquiterpene with countless biological properties. Appl. Sci. 2019, 9, 5420. [Google Scholar] [CrossRef][Green Version]
  45. Mao, J.; Yi, M.; Wang, R.; Huang, Y.; Chen, M. Protective effects of costunolide against D-galactosamine and lipopolysaccharide-induced acute liver injury in mice. Front. Pharmacol. 2018, 9, 1469. [Google Scholar] [CrossRef][Green Version]
  46. Kim, D.Y.; Choi, B.Y. Costunolide—A bioactive sesquiterpene lactone with diverse therapeutic potential. Int. J. Mol. Sci. 2019, 20, 2926. [Google Scholar] [CrossRef] [PubMed][Green Version]
  47. Park, J.; Park, J.; Leem, Y.; Kim, D.; Kim, H. NQO1 mediates the anti-inflammatory effects of Nootkatone in lipopolysaccharide-induced neuroinflammation by modulating the AMPK Signaling pathway. Free Radic. Biol. Med. 2021, 164, 354–368. [Google Scholar] [CrossRef] [PubMed]
  48. Kurdi, A.; Hassan, K.; Venkataraman, B.; Rajesh, M. Nootkatone confers hepatoprotective and anti-fibrotic actions in a murine model of liver fibrosis by suppressing oxidative stress, inflammation, and apoptosis. J. Biochem. Mol. Toxicol. 2017, 32, e22017. [Google Scholar] [CrossRef]
  49. Xu, J.; Zhang, L.; Jiang, R.; Hu, K.; Hu, D.; Liao, C.; Jiang, S.; Yang, Y.; Huang, J.; Tang, L.; et al. Nicotinamide improves NAD+ levels to protect against acetaminophen-induced acute liver injury in mice. Hum. Exp. Toxicol. 2021, 40, 1938–1946. [Google Scholar] [CrossRef]
  50. Lappas, M.; Permezel, M. The anti-inflammatory and antioxidative effects of nicotinamide, a vitamin B3 derivative, are elicited by FoxO3 in human gestational tissues: Implications for preterm birth. J. Nutr. Biochem. 2011, 22, 1195–1201. [Google Scholar] [CrossRef]
  51. Eidi, A.; Mortazavi, P.; Tehrani, M.E.; Rohani, A.H.; Safi, S. Hepatoprotective effects of pantothenic acid on carbon tetrachloride-induced toxicity in rats. EXCLI J. 2012, 11, 748–759. [Google Scholar] [PubMed]
  52. Roh, T.; De, U.; Lim, S.K.; Kim, M.K.; Choi, S.M.; Lim, D.S. Detoxifying effect of pyridoxine on acetaminophen-induced hepatotoxicity via suppressing oxidative stress injury. Food Chem. Toxicol. 2018, 114, 11–22. [Google Scholar] [CrossRef] [PubMed]
  53. Bird, R.P. The emerging role of vitamin B6 in inflammation and carcinogenesis. Adv. Food Nutr. Res. 2018, 83, 151–194. [Google Scholar] [CrossRef]
  54. Hu, X.; Wang, M.; Pan, Y.; Xie, Y.; Han, J.; Zhang, X.; Niayale, R.; He, H.; Li, Q.; Zhao, T.; et al. Anti-inflammatory effect of Astragalin and chlorogenic acid on Escherichia coli-induced inflammation of sheep endometrial epithelium cells. Front. Vet. Sci. 2020, 7, 201. [Google Scholar] [CrossRef]
  55. Riaz, A.; Rasul, A.; Hussain, G.; Zahoor, M.K.; Jabeen, F.; Subhani, Z.; Younis, T.; Ali, M.; Sarfraz, I.; Selamoglu, Z. Astragalin: A bioactive phytochemical with potential therapeutic activities. Adv. Pharmacol. Sci. 2018, 2018, 9794625. [Google Scholar] [CrossRef][Green Version]
  56. Liu, Q.; Pan, R.; Ding, L.; Zhang, F.; Hu, L.; Ding, B.; Zhu, L.; Xia, Y.; Dou, X. Rutin exhibits hepatoprotective effects in a mouse model of non-alcoholic fatty liver disease by reducing hepatic lipid levels and mitigating lipid-induced oxidative injuries. Int. Immunopharmacol. 2017, 49, 132–141. [Google Scholar] [CrossRef]
  57. Yoo, H.; Ku, S.; Baek, Y.; Bae, J. Anti-inflammatory effects of Rutin on HMGB1-induced inflammatory responses in vitro and in vivo. Inflamm. Res. 2013, 63, 197–206. [Google Scholar] [CrossRef] [PubMed]
  58. Murphy, R.C. Tandem Mass Spectrometry of Lipids: Molecular Analysis of Complex Lipids, 1st ed.; Royal Society of Chemistry: Cambridge, UK, 2015. [Google Scholar]
  59. Kuncoro, H.; Farabi, K.; Rijai, L. Steroids and isoquercetin from Lygodium microphyllum. J. Appl. Pharm. Sci. 2017, 7, 136–141. [Google Scholar] [CrossRef][Green Version]
  60. Kowalski, S. Changes of antioxidant activity and formation of 5-hydroxymethylfurfural in honey during thermal and microwave processing. Food Chem. 2013, 141, 1378–1382. [Google Scholar] [CrossRef]
  61. Ghaderi, S.; Gholipour, P.; Komaki, A.; Salehi, I.; Rashidi, K.; Esmaeil Khoshnam, S.; Rashno, M. P-coumaric acid ameliorates cognitive and non-cognitive disturbances in a rat model of Alzheimer’s disease: The role of oxidative stress and inflammation. Int. Immunopharmacol. 2022, 112, 109295. [Google Scholar] [CrossRef]
  62. Aparna, V.; Dileep, K.V.; Mandal, P.K.; Karthe, P.; Sadasivan, C.; Haridas, M. Anti-inflammatory property of n-hexadecanoic acid: Structural evidence and kinetic assessment. Chem. Biol. Drug Des. 2012, 80, 434–439. [Google Scholar] [CrossRef] [PubMed]
  63. Yogeswari, S.; Ramalakshmy, S.; Neelavathy, R.; Muthumary, J. Identification and comparative studies of different volatile fractions from Monochaetia kansensis by GCMS. Glob. J. Pharmacol. 2012, 6, 65–71. [Google Scholar]
  64. Breijyeh, Z.; Jubeh, B.; Karaman, R. Resistance of Gram-negative bacteria to current antibacterial agents and approaches to resolve it. Molecules 2020, 16, 1340. [Google Scholar] [CrossRef][Green Version]
  65. Urbanek, A.; Szadziewski, R.; Stepnowski, P.; Boros-Majewska, J.; Gabriel, I.; Dawgul, M.; Kamysz, W.; Sosnowska, D.; Gołębiowski, M. Composition and antimicrobial activity of fatty acids detected in the hygroscopic secretion collected from the secretory setae of larvae of the biting midge Forcipomyia nigra (Diptera: Ceratopogonidae). J. Insect Physiol. 2012, 58, 1265–1276. [Google Scholar] [CrossRef] [PubMed]
  66. Azam, F.; Chaudhry, B.A.; Ijaz, H.; Qadir, M.I. Caffeoyl-β-D-glucopyranoside and 1,3-dihydroxy-2-tetracosanoylamino-4-(e)-nonadecene isolated from ranunculus muricatus exhibit antioxidant activity. Sci. Rep. 2019, 9, 15613. [Google Scholar] [CrossRef][Green Version]
  67. Wlodarska, M.; Luo, C.; Kolde, R.; d’Hennezel, E.; Annand, J.W.; Heim, C.E.; Krastel, P.; Schmitt, E.K.; Omar, A.S.; Creasey, E.A.; et al. Indoleacrylic acid produced by commensal peptostreptococcus species suppresses inflammation. Cell Host Microbe. 2017, 22, 25–37. [Google Scholar] [CrossRef][Green Version]
  68. Li, S.; Zhang, Z.; Cain, A.; Wang, B.; Long, M.; Taylor, J. Antifungal activity of Camptothecin, trifolin, and hyperoside isolated from Camptotheca acuminata. J. Agric. Food. Chem. 2005, 53, 32–37. [Google Scholar] [CrossRef]
  69. Das, U.N. Arachidonic acid and other unsaturated fatty acids and some of their metabolites function as endogenous antimicrobial molecules: A Review. J. Adv. Res. 2018, 11, 57–66. [Google Scholar] [CrossRef]
Table 1. Compounds identified in LM aqueous extract by positive mode of analysis.
Table 1. Compounds identified in LM aqueous extract by positive mode of analysis.
NameR. Time (min)FormulaMass Error (ppm)Calc. Molecular MassDatabaseMatching Score (MzCloud)/
FISh Score (Chemspider)
Pyroglutamic acid1.04C5H7NO30.56129.0427mzCloud95.6Amino acid
Succinylproline4.79C9H13NO50.45215.0795ChemSpider50.0Amino acid
3-Indoleacrylic acid6.35C11H9NO20.15187.0634mzCloud94.3Aromatic
Phenylpropiolic acid5.64C9H6O2−0.86146.0367ChemSpider50.0Aromatic
9S,13R-12-Oxophytodienoic acid8.13C18H28O3−0.84292.2036mzCloud92.1Cyclic ketone
Jasmonic acid8.51C12H18O3−0.28210.1255ChemSpider66.3Cyclic ketone
Maltol3.74C6H6O30.85126.0318mzCloud99.3Cyclic ketone
Vomifoliol4.97C13H20O3−0.44224.1412ChemSpider69.9Cyclic ketone
(12S)-12-Hydroxy-16-heptadecynoic acid10.81C17H30O3−0.10282.2195ChemSpider53.9Fatty acyl
(2E)-6-Hydroxy-2,6-dimethyl-2,7-octadienoic acid5.09C10H16O30.48184.1100ChemSpider54.6Fatty acyl
1-[(2-Hydroxyethyl)amino]-2-dodecanol21.44C14H31NO2−0.33245.2354ChemSpider55.6Fatty acyl
11-Methoxy-3,7,11-trimethyl-2,4-dodecadienoic acid8.43C16H28O3−0.12268.2038ChemSpider65.9Fatty acyl
13-Hydroxy-9,11,15-octadecatrienoic acid8.52C18H30O3−0.40294.2194mzCloud85.4Fatty acyl
4-Oxo-dodecanedioic acid6.44C12H20O5−0.52244.1310mzCloud80.2Fatty acyl
Arachidonic acid7.01C20H32O2−0.98304.2399ChemSpider90.5Fatty acyl
Levulinic acid1.10C5H8O31.96116.0476ChemSpider62.5Fatty acyl
Palmitoleyl oleate20.98C34H64O2−0.03504.4906ChemSpider85.8Fatty acyl
Parinaric acid11.45C18H28O2−0.74276.2087ChemSpider85.6Fatty acyl
Traumatin5.76C12H20O3−0.11212.1412ChemSpider56.4Fatty acyl
2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4-oxo-4H-chromen-3-yl 6-O-(carboxyacetyl)-β-D-threo-hexopyranoside5.84C24H22O150.78550.0963mzCloud99.0Flavonoid
Luteolin 7-O-malonylglucoside6.22C24H22O140.31534.1011ChemSpider62.5Flavonoid
Quercetin 3-O-rhamnoside-7-O-glucoside5.54C27H30O16−0.33610.1532mzCloud98.2Flavonoid
Caffeic acid4.10C9H8O4−0.62180.0422mzCloud99.2Phenolic
Steroidal compound19.77C29H46O−0.65410.3546ChemSpider62.3Steroid
Steroidal compound8.07C18H26O2−0.98274.1930mzCloud93.8Steroid
Caryophyllene oxide6.69C15H24O−0.85220.1825mzCloud80.2Terpenoid
Perillic acid5.56C10H14O2−0.54166.0993ChemSpider54.0Terpenoid
Nicotinamide1.05C6H6N2O1.13122.0482mzCloud97.4Vitamin B
Pantothenic acid3.50C9H17NO5−0.06219.1107ChemSpider95.2Vitamin B
Pyridoxine1.01C8H11NO3−0.14169.0739mzCloud96.2Vitamin B
Table 2. Compounds identified in LM aqueous extract by negative mode of analysis.
Table 2. Compounds identified in LM aqueous extract by negative mode of analysis.
NameR. Time (min)FormulaMass Error (ppm)Calc. Molecular MassDatabaseMatching Score (MzCloud)/
FISh Score (Chemspider)
(15Z)-9,12,13-Trihydroxy-15-octadecenoic acid8.53C18H34O50.53330.2408mzCloud88.3Fatty acyl
12,13-Dihydroxyoctadec-9-enoic acid10.75C18H34O40.20314.2458mzCloud90.0Fatty acyl
13-Hydroxy-9,11,15-octadecatrienoic acid11.46C18H30O30.83294.2197mzCloud89.4Fatty acyl
13-Hydroxy-9,11-octadecadienoic acid12.06C18H32O30.33296.2352mzCloud85.5Fatty acyl
16-Hydroxyhexadecanoic acid14.12C16H32O30.36272.2352mzCloud87.5Fatty acyl
9-Hydroperoxy-10,12-octadecadienoic acid10.22C18H32O40.67312.2303mzCloud87.8Fatty acyl
Corchorifatty acid F8.09C18H32O50.70328.2252mzCloud97.9Fatty acyl
Dodecanedioic acid6.60C12H22O4−0.67230.1517mzCloud95.7Fatty acyl
5,7-Dihydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-3-yl 6-O-(6-deoxyhexopyranosyl)hexopyranoside5.86C27H30O150.70594.1589mzCloud88.0Flavonoid
Table 3. Compounds identified in the aqueous extract of LM by GC-MS.
Table 3. Compounds identified in the aqueous extract of LM by GC-MS.
No.Compound NameMolecular FormulaMolecular WeightArea (%)RT
3.2-furancarboxaldehyde, 5-methyl-C6H6O2110.110.1611.4392
4.Benzyl alcoholC7H8O108.140.2814.6725
5.Acetic acid, phenylmethyl esterC9H10O2150.170.2020.7917
7.p-Coumaric acidC9H8O3164.160.3446.3873
9.n-Hexadecanoic acidC16H32O2256.420.1851.7683
Table 4. Inhibition zone produced by LM against test bacteria.
Table 4. Inhibition zone produced by LM against test bacteria.
Inhibition Zone (mm)
SamplesConcentration (mg/mL)S. aureusB. cereusE. coliSalmonella sp.
LM50015.0 ± 3.1---
10013.0 ± 1.7---
Gentamicin10 µg/disc23.0 ± 0.625.0 ± 1.524.0 ± 1.028.0 ± 0.6
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Teoh, W.Y.; Yong, Y.S.; Razali, F.N.; Stephenie, S.; Dawood Shah, M.; Tan, J.K.; Gnanaraj, C.; Mohd Esa, N. LC-MS/MS and GC-MS Analysis for the Identification of Bioactive Metabolites Responsible for the Antioxidant and Antibacterial Activities of Lygodium microphyllum (Cav.) R. Br. Separations 2023, 10, 215.

AMA Style

Teoh WY, Yong YS, Razali FN, Stephenie S, Dawood Shah M, Tan JK, Gnanaraj C, Mohd Esa N. LC-MS/MS and GC-MS Analysis for the Identification of Bioactive Metabolites Responsible for the Antioxidant and Antibacterial Activities of Lygodium microphyllum (Cav.) R. Br. Separations. 2023; 10(3):215.

Chicago/Turabian Style

Teoh, Wuen Yew, Yoong Soon Yong, Faizan Naeem Razali, Sarah Stephenie, Muhammad Dawood Shah, Jen Kit Tan, Charles Gnanaraj, and Norhaizan Mohd Esa. 2023. "LC-MS/MS and GC-MS Analysis for the Identification of Bioactive Metabolites Responsible for the Antioxidant and Antibacterial Activities of Lygodium microphyllum (Cav.) R. Br." Separations 10, no. 3: 215.

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop