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Phytochemical Screening and Therapeutic Potential Evaluation from the leaves of Alangium chinense (Lour.) Harms to Validate Traditional Medicinal Knowledge

L
Lakshmikanta Khundrakpam1,*
A
Ajit Kumar Ngangbam1
B
Bijayalakshmi Devi Nongmaithem1
L
Laiphrakpam Pinky Chanu1
L
Laishram Lenin Singh1
U
Urikhimbam Bipinchandra Singh1
K
Kayenpaibam Monorama Devi1
1School of Biological Sciences, Manipur International University, Imphal-795 140, Manipur, India.

ABSTRACT

Background: Alangium chinense is a traditionally important medicinal plant among the indigenous community of Manipur, India. However, its bioactive profile has not been scientifically investigated to date. This study presents the preliminary chemical profiling of A. chinense leaves using LC-MS and GC-MS.

Methods: Chemical profiling of A. chinense leaves was conducted using solvent extraction methods, followed by LC-MS and GC-MS analysis to identify and evaluate the bioactive compounds.

Result: This study reveals the presence of several phytochemicals including sinapic acid, leonurine, pyridinoline, kyotorphin, goyaglycoside C, (R)-hydnocarpic acid, pentanoic acid, 5-hydroxy-2,4-di-t-butylphenyl, 2-methylhexacosane, tetracosane, nonacosane, phenol, 2,4-bis (1,1-dimethylethyl)- phosphite (3:1) and 9,12-octadecadienoic acid many of which are known for their antioxidant, anticancer, anti-inflammatory, antimicrobial, neuroprotective and antidiabetic properties. A novel compound, medicanine and several other metabolites with unknown bioactivities were also detected during the study, which highlights the need for further studies to explore their potential bioactive roles. Overall, this research highlights the potential of A. chinense as a promising candidate for prospective drug discovery, dermo-cosmetics and functional foods, while promoting conservation of ethno-medicine and biodiversity.

INTRODUCTION

Humans have a long history of using herbs and shrubs as a source of medicine since ancient times. Ancient civilisations relied significantly on plants that were concurrently synergistic for sustenance and the treatment of certain illnesses. India is not only a land of diversity, but also stands out for its rich abundance of an immense variety of medicinal plants. The Botanical Survey of India stated that around 45,000 plant species were native to India, of which around 8,000 species were used as a source of medicine by traditional practitioners (Ved et al., 2001). Nearly 50% of India’s total plant diversity flourishes in the Northeast region (Dutta and Dutta, 2005). Manipur is one of the Northeast states of India, encircled by nine enchanting hill ranges, covered with a variety of lush green species, forming an oval-shaped valley at the heart of the state. Manipur can be termed the “Concentrated biodiversity hotspot” of India and is renowned for its diverse flora and fauna, which are loaded with significant amounts of potent bioactive compounds. The indigenous community of Manipur shares a symbiotic relationship with the biome’s biodiversity and the utilisation of plants as medicine for treating various infectious and non-infectious maladies has been practiced since ancient times. Approximately 1,500 species of medicinal plants native to Manipur have been recorded to date (Dutta et al., 2023). Plant-derived compounds are also emerging as a sustainable and eco-friendly alternative to synthetic chemicals, offering antimicrobial and protective functions (Malagi et al., 2024). Plant-derived phytochemicals neutralise free radicals and oxidative stress, thereby preventing cellular damage, with flavonoids and phenolic-rich extracts exhibiting significant therapeutic potential (Pant et al., 2024). Medicinal plants with high levels of secondary metabolites have been extensively investigated for their antimicrobial properties. Their diverse phytochemical composition strengthens their biological efficacy, supporting their use in ethnomedicine (Singh et al., 2023).
       
Alangium spp. distinguishes itself as one of the potent medicinal plants exhibiting a broad spectrum of health benefits, notably, antidiabetic, anticancer, diuretic, anti-inflammatory, anti-microbial, etc. (Shetty, 2003; Xavier et al., 2005; Jain et al., 2010). A. chinense (Lour.) Harms (Manipuri name Kokal) is a tropical plant with a rich ethno-pharmacological history and perceived protective properties. However, due to the lack of in-depth research and insufficient scientific investigation, the potential of the bioactive compounds in A. chinense remains unknown in the region’s population. A comprehensive survey was carried out during March–June, 2024 across different regions of Manipur to gather therapeutic insights from local communities. Local respondents reported the traditional use of leaves during solar eclipses as a protective measure against perceived harmful radiation; however, this practice remains anecdotal and lacks scientific validation (Personal communication). This study presents the preliminary chemical profiling of A. chinense leaves using LC-MS and GC-MS, providing baseline phytochemical evidence to support traditional medicinal knowledge, forming a foundation for future bioactivity-guided and in vivo investigations.

MATERIALS AND METHODS

The study was conducted at the Department of Biotechnology, Manipur International University, from November 2024 to April 2025. Samples of A. chinense leaves were collected from Uchekon, Manipur, India, in October 2024. Leaf samples were collected from three independent plants (n = 3 biological replicates). A qualified taxonomist authenticated the plant material and a voucher specimen has been deposited at the Herbarium of Manipur International University (Voucher No.: MIU-AC-2024-01). The extraction yields were calculated as percentages (w/w) relative to dried plant material, yielding approximately 8.6% (methanol extract) and 6.4% (chloroform extract). Instrument calibration was performed using standard tuning mixtures supplied by the manufacturer before analysis to ensure mass accuracy and reproducibility.
       
To prepare a sequential lipophilic solution (chloroform) and polar solvent (methanol) extracts, the freshly collected leaves of A. chinense were stored at room temperature and processed within 24 hours to preserve their chemical integrity. The leaves were gently rinsed with sterile distilled water to remove surface contaminants, excised using sterile scissors and dried. The dried leaves were homogenised to create a uniform mixture and subsequently subjected to solvent extraction. Extractions were performed using chloroform to isolate lipophilic compounds and methanol to extract polar metabolites. Both extracts were filtered to remove particulate matter. The resulting extracts were analysed using LC-MS and GC-MS to profile and quantify the bioactive compounds. Strict Laboratory protocols were followed to prevent cross-contamination between samples and all equipment was thoroughly sterilised to ensure the reliability of the results.
 
Extraction of lipophilic and polar compounds from the leaf tissues of A. chinense
 
The fresh leaf tissues of A. chinense (25 g) were ground using a mortar and pestle to facilitate the disruption of cellular structures and enhance the release of bioactive compounds. The finely ground leaf tissues were then immersed in 100 ml each of HPLC-grade (Sigma-Aldrich, St. Louis, MO, USA) chloroform and methanol solvents for 48 hours. Finally, the solvents were decanted and the samples were macerated overnight in freshly replenished solvents at 4°C in a refrigerator (Videocon 190 L, Videocon, Mumbai, India). The entire extraction process was conducted under controlled conditions to minimise the degradation of sensitive compounds and ensure optimal extraction efficiency. The extracts were filtered through Whatman No. 1 filter paper (Whatman, Buckinghamshire, UK) to eliminate particulate matter. The filtrate was then concentrated under reduced pressure using a rotary evaporator (Buchi, Flawil, Switzerland) set to 40°C and 150 mb pressure to evaporate the solvents. The dried leaf extracts (10g) were reconstituted in 15 ml of the respective solvents and then transferred to pre-weighed glass vials. The final weight of the extracts was recorded and stored at -20°C in a Blue Star freezer for subsequent analysis. The profiling of bioactive compounds from the leaves of A. chinense was carried out by LC-MS with an Agilent 1260 Infinity II/LC-MSD iQ system. The volatile compounds were identified by GC-MS on an Agilent 7890A GC (Agilent Technologies, Palo Alto, CA, USA) coupled with an MS (5977B VL MSD, Agilent Technologies).
 
LC-MS analysis of leaf extracts of A. chinense
 
The chloroform and methanol extracts were subjected to LC-MS analysis for the identification of bioactive compounds. A 2.1 × 50 mm C18 column was used and gradient elution was applied over 7 minutes, with a mobile phase starting at 5:95 acetonitrile/water containing 0.1% formic acid, gradually transitioning to 95:5 acetonitrile/water with 0.1% formic acid. The LC-MS analysis was performed on an Agilent 1260 Infinity II/LCMSD iQ system (Agilent technologies) following the protocols outlined by Nongmaithem et al., (2017). Mass spectrometry data were acquired within the range of 100-800 m/z in both ESI+ and ESI- ionisation modes, with an injection volume of 10 μL for all samples. Electrospray ionisation was operated with a capillary voltage of 3500 V, drying gas temperature of 325°C, gas flow rate of 10 L min-1 and nebuliser pressure of 35 psi. The LC-MS data were analysed using Agilent ChemStation software and the inferred chemical formulas were cross-referenced with the National Library of Medicine, USA, to identify potential bioactive metabolites. 
 
GC-MS analysis of leaf extracts of A. chinense
 
The chloroform and methanol leaf extracts of A. chinense were subjected to GC-MS (60 min) for the profiling of volatile organic compounds (VOCs) using an Agilent 7890A gas chromatograph, interfaced with a 5977B VL MSD mass spectrometer (Agilent Technologies), adopting the procedures provided by Nongmaithem et al., (2017). The GC temperature was initiated at 40°C, held for 5 min, increased to 120°C (2 min hold) with a ramp rate of 3°C/min and finally set to 250°C at the rate of 8°C/min for 10 min. The volatile compounds were identified and characterised by comparing the obtained mass spectra with those listed in the National Institute of Standards and Technology (NIST) 2017 library database.

RESULTS AND DISCUSSION

The LC-MS and GC-MS analyses identified 29 major metabolites that had not been previously documented in the leaves of A. chinense. Our research findings highlight the metabolic complexity and phytochemical diversity of A. chinense, showcasing its potential pharmaceutical applications. The LC-MS and GC-MS chromatograms (Methanolic and Chloroform extracts) are shown in Fig 1 and 2, respectively. The compounds identified via LC-MS and GC-MS (Methanolic and chloroform extracts) were presented in Table 1 and 2, respectively, along with their chemical formula, retention time, major ion, score and bioactive properties. Compound identification was based on spectral matching scores and accurate mass comparison; however, these annotations remain putative and require structural confirmation using authentic standards or NMR analysis. The LC-MS and GC-MS analyses revealed the presence of several pharmacologically significant bioactive metabolites, which lay a foundation for further exploration of A. chinense in drug discovery and its potential as a promising source of novel bioactive compounds.

Fig 1: LC-MS chromatograms of A. chinense leaf extracts.



Fig 2: GC-MS chromatograms of A. chinense leaf extracts.



Table 1: Chemical constituents of A. chinense (Lour.) harms leaves-methanol and chloroform extracts using LC-MS.



Table 2: Chemical constituents of A. chinense (Lour.) harms leaves-chloroform and methanolic extracts using GC-MS.


       
While several detected compounds have been shown to exhibit bioactivity in previous studies, their effects in A. chinense extracts remain speculative and require experimental validation. Some of the major bioactive compounds detected using LC-MS include sinapic acid which is known for strong antioxidant and anti-inflammatory properties (Nićiforović and Abramovič, 2014; Yun et al., 2008), Leonurine has been shown to have both cardioprotective and neuroprotective effects (Liu et al., 2009; Qi et al., 2010), 11-amino-undecanoic acid for its anti-inflammatory and antioxidant properties (Ikeda et al., 2008), leonuriside A for its antioxidant properties (Sugaya et al., 1998),  corchoionol C 9-glucoside, an antihyperglycemic and antioxidant (Lestari et al., 2024) and icaceine which exhibits anticonvulsant properties (Dixit and Reddy, 2017). Along with these major compounds, pyridinoline is also known to be a collagen cross-linker, suggesting possible dermatological and osteological uses (Tapia-Vázquez et al., 2025).
       
Moreover, the results support the phyto-therapeutic potential and functional properties as well as their traditional medicinal applications. The GC-MS analysis revealed the presence of various chemical compounds, which show anti-inflammatory, antibacterial and antifungal properties. Interestingly, tetracosane and nonacosane showed apoptotic effects in cell lines and possess anti-mutagenic properties, suggesting potential anticancer properties, which could be used in cancer prevention and related therapeutic applications (Uddin et al., 2012; Kalsum et al., 2016). Besides these compounds, phenol, 2,4-bis (1,1-dimethylethyl)-, phosphite (3:1) and 9,12-octadecadienoic acid (Z,Z), also known as linoleic acid, detected during this study exhibited antioxidant, antiviral, anticancer, antifungal, anti-enterococcal, anti-inflammatory, anticancer and antihistaminic activities (Patil and Singh, 2022; Hnbgu et al., 2021; Yan et al., 2024). Medicanine (Table 1) was tentatively annotated based on LC-MS spectral matching and is reported here as a putatively identified compound.
               
The chemical diversity of A. chinense shows its pharmacological potential, which may likely be developed due to environmental and microbial interactions in the biodiversity hotspot of Manipur. This aligns with earlier studies regarding the rich diversity of medicinal plants and ethno-pharmacology in this region (Mao and Roy, 2016).  A. chinense warrants further investigation through in vivo studies, bioactivity-guided fractionation and mechanism of action studies.  More studies on A. chinense chemical profile are needed, including cytotoxicity and other pharmacological verifications to confirm the efficacy, safety and validation of the plant-based compounds. Nevertheless, this research provides the foundation for future strategies in bioprospecting, integrative medicine and underlines the significance of respecting traditional knowledge systems and biological diversity. The observed metabolite diversity may reflect adaptive biochemical responses to environmental stressors, microbial interactions and ecological pressures characteristic of the Manipur biodiversity hotspot. 

CONCLUSION

This study provides the preliminary phytochemical data that supports the traditional medicinal use of A. chinense through LC-MS and GC-MS. The presence of several bioactive compounds such as sinapic acid, leonurine, kyotorphin and hydnocarpic acid highlights its possible therapeutic activities as an antioxidant, anti-inflammatory, antimicrobial, neuroprotective and antidiabetic agent. This study further supports the ethnobotanical knowledge of the indigenous communities of Manipur and highlights the importance of conserving and exploring traditional remedies. Moreover, the study opens new research areas for bioprospecting of A. chinense as a promising source of functional ingredients for pharmaceuticals, nutraceuticals and cosmeceuticals.  Bioassay-guided fractionation, in vivo studies and clinical trials will be necessary to fully harness the pharmaceutical potential of this underutilised A. chinense.

ACKNOWLEDGEMENT

The authors sincerely acknowledge Mr. M. Joychandra, Ph.D., research scholar, for his valuable support during the research work. The laboratory facilities and analytical instrumentation provided at Manipur International University were instrumental in successfully conducting this research work.

CONFLICT OF INTEREST

The authors declare that no conflict of interest was reported regarding the publication of this article. 

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    Full Research Article

    Phytochemical Screening and Therapeutic Potential Evaluation from the leaves of Alangium chinense (Lour.) Harms to Validate Traditional Medicinal Knowledge

    L
    Lakshmikanta Khundrakpam1,*
    A
    Ajit Kumar Ngangbam1
    B
    Bijayalakshmi Devi Nongmaithem1
    L
    Laiphrakpam Pinky Chanu1
    L
    Laishram Lenin Singh1
    U
    Urikhimbam Bipinchandra Singh1
    K
    Kayenpaibam Monorama Devi1
    1School of Biological Sciences, Manipur International University, Imphal-795 140, Manipur, India.

    ABSTRACT

    Background: Alangium chinense is a traditionally important medicinal plant among the indigenous community of Manipur, India. However, its bioactive profile has not been scientifically investigated to date. This study presents the preliminary chemical profiling of A. chinense leaves using LC-MS and GC-MS.

    Methods: Chemical profiling of A. chinense leaves was conducted using solvent extraction methods, followed by LC-MS and GC-MS analysis to identify and evaluate the bioactive compounds.

    Result: This study reveals the presence of several phytochemicals including sinapic acid, leonurine, pyridinoline, kyotorphin, goyaglycoside C, (R)-hydnocarpic acid, pentanoic acid, 5-hydroxy-2,4-di-t-butylphenyl, 2-methylhexacosane, tetracosane, nonacosane, phenol, 2,4-bis (1,1-dimethylethyl)- phosphite (3:1) and 9,12-octadecadienoic acid many of which are known for their antioxidant, anticancer, anti-inflammatory, antimicrobial, neuroprotective and antidiabetic properties. A novel compound, medicanine and several other metabolites with unknown bioactivities were also detected during the study, which highlights the need for further studies to explore their potential bioactive roles. Overall, this research highlights the potential of A. chinense as a promising candidate for prospective drug discovery, dermo-cosmetics and functional foods, while promoting conservation of ethno-medicine and biodiversity.

    INTRODUCTION

    Humans have a long history of using herbs and shrubs as a source of medicine since ancient times. Ancient civilisations relied significantly on plants that were concurrently synergistic for sustenance and the treatment of certain illnesses. India is not only a land of diversity, but also stands out for its rich abundance of an immense variety of medicinal plants. The Botanical Survey of India stated that around 45,000 plant species were native to India, of which around 8,000 species were used as a source of medicine by traditional practitioners (Ved et al., 2001). Nearly 50% of India’s total plant diversity flourishes in the Northeast region (Dutta and Dutta, 2005). Manipur is one of the Northeast states of India, encircled by nine enchanting hill ranges, covered with a variety of lush green species, forming an oval-shaped valley at the heart of the state. Manipur can be termed the “Concentrated biodiversity hotspot” of India and is renowned for its diverse flora and fauna, which are loaded with significant amounts of potent bioactive compounds. The indigenous community of Manipur shares a symbiotic relationship with the biome’s biodiversity and the utilisation of plants as medicine for treating various infectious and non-infectious maladies has been practiced since ancient times. Approximately 1,500 species of medicinal plants native to Manipur have been recorded to date (Dutta et al., 2023). Plant-derived compounds are also emerging as a sustainable and eco-friendly alternative to synthetic chemicals, offering antimicrobial and protective functions (Malagi et al., 2024). Plant-derived phytochemicals neutralise free radicals and oxidative stress, thereby preventing cellular damage, with flavonoids and phenolic-rich extracts exhibiting significant therapeutic potential (Pant et al., 2024). Medicinal plants with high levels of secondary metabolites have been extensively investigated for their antimicrobial properties. Their diverse phytochemical composition strengthens their biological efficacy, supporting their use in ethnomedicine (Singh et al., 2023).
           
    Alangium     spp. distinguishes itself as one of the potent medicinal plants exhibiting a broad spectrum of health benefits, notably, antidiabetic, anticancer, diuretic, anti-inflammatory, anti-microbial, etc. (Shetty, 2003; Xavier et al., 2005; Jain et al., 2010). A. chinense (Lour.) Harms (Manipuri name Kokal) is a tropical plant with a rich ethno-pharmacological history and perceived protective properties. However, due to the lack of in-depth research and insufficient scientific investigation, the potential of the bioactive compounds in A. chinense remains unknown in the region’s population. A comprehensive survey was carried out during March–June, 2024 across different regions of Manipur to gather therapeutic insights from local communities. Local respondents reported the traditional use of leaves during solar eclipses as a protective measure against perceived harmful radiation; however, this practice remains anecdotal and lacks scientific validation (Personal communication). This study presents the preliminary chemical profiling of A. chinense leaves using LC-MS and GC-MS, providing baseline phytochemical evidence to support traditional medicinal knowledge, forming a foundation for future bioactivity-guided and in vivo investigations.

    MATERIALS AND METHODS

    The study was conducted at the Department of Biotechnology, Manipur International University, from November 2024 to April 2025. Samples of A. chinense leaves were collected from Uchekon, Manipur, India, in October 2024. Leaf samples were collected from three independent plants (n = 3 biological replicates). A qualified taxonomist authenticated the plant material and a voucher specimen has been deposited at the Herbarium of Manipur International University (Voucher No.: MIU-AC-2024-01). The extraction yields were calculated as percentages (w/w) relative to dried plant material, yielding approximately 8.6% (methanol extract) and 6.4% (chloroform extract). Instrument calibration was performed using standard tuning mixtures supplied by the manufacturer before analysis to ensure mass accuracy and reproducibility.
           
    To prepare a sequential lipophilic solution (chloroform) and polar solvent (methanol) extracts, the freshly collected leaves of A. chinense were stored at room temperature and processed within 24 hours to preserve their chemical integrity. The leaves were gently rinsed with sterile distilled water to remove surface contaminants, excised using sterile scissors and dried. The dried leaves were homogenised to create a uniform mixture and subsequently subjected to solvent extraction. Extractions were performed using chloroform to isolate lipophilic compounds and methanol to extract polar metabolites. Both extracts were filtered to remove particulate matter. The resulting extracts were analysed using LC-MS and GC-MS to profile and quantify the bioactive compounds. Strict Laboratory protocols were followed to prevent cross-contamination between samples and all equipment was thoroughly sterilised to ensure the reliability of the results.
     
    Extraction of lipophilic and polar compounds from the leaf tissues of A. chinense
     
    The fresh leaf tissues of A. chinense (25 g) were ground using a mortar and pestle to facilitate the disruption of cellular structures and enhance the release of bioactive compounds. The finely ground leaf tissues were then immersed in 100 ml each of HPLC-grade (Sigma-Aldrich, St. Louis, MO, USA) chloroform and methanol solvents for 48 hours. Finally, the solvents were decanted and the samples were macerated overnight in freshly replenished solvents at 4°C in a refrigerator (Videocon 190 L, Videocon, Mumbai, India). The entire extraction process was conducted under controlled conditions to minimise the degradation of sensitive compounds and ensure optimal extraction efficiency. The extracts were filtered through Whatman No. 1 filter paper (Whatman, Buckinghamshire, UK) to eliminate particulate matter. The filtrate was then concentrated under reduced pressure using a rotary evaporator (Buchi, Flawil, Switzerland) set to 40°C and 150 mb pressure to evaporate the solvents. The dried leaf extracts (10g) were reconstituted in 15 ml of the respective solvents and then transferred to pre-weighed glass vials. The final weight of the extracts was recorded and stored at -20°C in a Blue Star freezer for subsequent analysis. The profiling of bioactive compounds from the leaves of A. chinense was carried out by LC-MS with an Agilent 1260 Infinity II/LC-MSD iQ system. The volatile compounds were identified by GC-MS on an Agilent 7890A GC (Agilent Technologies, Palo Alto, CA, USA) coupled with an MS (5977B VL MSD, Agilent Technologies).
     
    LC-MS analysis of leaf extracts of A. chinense
     
    The chloroform and methanol extracts were subjected to LC-MS analysis for the identification of bioactive compounds. A 2.1 × 50 mm C18 column was used and gradient elution was applied over 7 minutes, with a mobile phase starting at 5:95 acetonitrile/water containing 0.1% formic acid, gradually transitioning to 95:5 acetonitrile/water with 0.1% formic acid. The LC-MS analysis was performed on an Agilent 1260 Infinity II/LCMSD iQ system (Agilent technologies) following the protocols outlined by Nongmaithem et al., (2017). Mass spectrometry data were acquired within the range of 100-800 m/z in both ESI+ and ESI- ionisation modes, with an injection volume of 10 μL for all samples. Electrospray ionisation was operated with a capillary voltage of 3500 V, drying gas temperature of 325°C, gas flow rate of 10 L min-1 and nebuliser pressure of 35 psi. The LC-MS data were analysed using Agilent ChemStation software and the inferred chemical formulas were cross-referenced with the National Library of Medicine, USA, to identify potential bioactive metabolites. 
     
    GC-MS analysis of leaf extracts of A. chinense
     
    The chloroform and methanol leaf extracts of A. chinense were subjected to GC-MS (60 min) for the profiling of volatile organic compounds (VOCs) using an Agilent 7890A gas chromatograph, interfaced with a 5977B VL MSD mass spectrometer (Agilent Technologies), adopting the procedures provided by Nongmaithem et al., (2017). The GC temperature was initiated at 40°C, held for 5 min, increased to 120°C (2 min hold) with a ramp rate of 3°C/min and finally set to 250°C at the rate of 8°C/min for 10 min. The volatile compounds were identified and characterised by comparing the obtained mass spectra with those listed in the National Institute of Standards and Technology (NIST) 2017 library database.

    RESULTS AND DISCUSSION

    The LC-MS and GC-MS analyses identified 29 major metabolites that had not been previously documented in the leaves of A. chinense. Our research findings highlight the metabolic complexity and phytochemical diversity of A. chinense, showcasing its potential pharmaceutical applications. The LC-MS and GC-MS chromatograms (Methanolic and Chloroform extracts) are shown in Fig 1 and 2, respectively. The compounds identified via LC-MS and GC-MS (Methanolic and chloroform extracts) were presented in Table 1 and 2, respectively, along with their chemical formula, retention time, major ion, score and bioactive properties. Compound identification was based on spectral matching scores and accurate mass comparison; however, these annotations remain putative and require structural confirmation using authentic standards or NMR analysis. The LC-MS and GC-MS analyses revealed the presence of several pharmacologically significant bioactive metabolites, which lay a foundation for further exploration of A. chinense in drug discovery and its potential as a promising source of novel bioactive compounds.

    Fig 1: LC-MS chromatograms of A. chinense leaf extracts.



    Fig 2: GC-MS chromatograms of A. chinense leaf extracts.



    Table 1: Chemical constituents of A. chinense (Lour.) harms leaves-methanol and chloroform extracts using LC-MS.



    Table 2: Chemical constituents of A. chinense (Lour.) harms leaves-chloroform and methanolic extracts using GC-MS.


           
    While several detected compounds have been shown to exhibit bioactivity in previous studies, their effects in A. chinense extracts remain speculative and require experimental validation. Some of the major bioactive compounds detected using LC-MS include sinapic acid which is known for strong antioxidant and anti-inflammatory properties (Nićiforović and Abramovič, 2014; Yun et al., 2008), Leonurine has been shown to have both cardioprotective and neuroprotective effects (Liu et al., 2009; Qi et al., 2010), 11-amino-undecanoic acid for its anti-inflammatory and antioxidant properties (Ikeda et al., 2008), leonuriside A for its antioxidant properties (Sugaya et al., 1998),  corchoionol C 9-glucoside, an antihyperglycemic and antioxidant (Lestari et al., 2024) and icaceine which exhibits anticonvulsant properties (Dixit and Reddy, 2017). Along with these major compounds, pyridinoline is also known to be a collagen cross-linker, suggesting possible dermatological and osteological uses (Tapia-Vázquez et al., 2025).
           
    Moreover, the results support the phyto-therapeutic potential and functional properties as well as their traditional medicinal applications. The GC-MS analysis revealed the presence of various chemical compounds, which show anti-inflammatory, antibacterial and antifungal properties. Interestingly, tetracosane and nonacosane showed apoptotic effects in cell lines and possess anti-mutagenic properties, suggesting potential anticancer properties, which could be used in cancer prevention and related therapeutic applications (Uddin et al., 2012; Kalsum et al., 2016). Besides these compounds, phenol, 2,4-bis (1,1-dimethylethyl)-, phosphite (3:1) and 9,12-octadecadienoic acid (Z,Z), also known as linoleic acid, detected during this study exhibited antioxidant, antiviral, anticancer, antifungal, anti-enterococcal, anti-inflammatory, anticancer and antihistaminic activities (Patil and Singh, 2022; Hnbgu et al., 2021; Yan et al., 2024). Medicanine (Table 1) was tentatively annotated based on LC-MS spectral matching and is reported here as a putatively identified compound.
                   
    The chemical diversity of A. chinense shows its pharmacological potential, which may likely be developed due to environmental and microbial interactions in the biodiversity hotspot of Manipur. This aligns with earlier studies regarding the rich diversity of medicinal plants and ethno-pharmacology in this region (Mao and Roy, 2016).  A. chinense warrants further investigation through in vivo studies, bioactivity-guided fractionation and mechanism of action studies.  More studies on A. chinense chemical profile are needed, including cytotoxicity and other pharmacological verifications to confirm the efficacy, safety and validation of the plant-based compounds. Nevertheless, this research provides the foundation for future strategies in bioprospecting, integrative medicine and underlines the significance of respecting traditional knowledge systems and biological diversity. The observed metabolite diversity may reflect adaptive biochemical responses to environmental stressors, microbial interactions and ecological pressures characteristic of the Manipur biodiversity hotspot. 

    CONCLUSION

    This study provides the preliminary phytochemical data that supports the traditional medicinal use of A. chinense through LC-MS and GC-MS. The presence of several bioactive compounds such as sinapic acid, leonurine, kyotorphin and hydnocarpic acid highlights its possible therapeutic activities as an antioxidant, anti-inflammatory, antimicrobial, neuroprotective and antidiabetic agent. This study further supports the ethnobotanical knowledge of the indigenous communities of Manipur and highlights the importance of conserving and exploring traditional remedies. Moreover, the study opens new research areas for bioprospecting of A. chinense as a promising source of functional ingredients for pharmaceuticals, nutraceuticals and cosmeceuticals.  Bioassay-guided fractionation, in vivo studies and clinical trials will be necessary to fully harness the pharmaceutical potential of this underutilised A. chinense.

    ACKNOWLEDGEMENT

    The authors sincerely acknowledge Mr. M. Joychandra, Ph.D., research scholar, for his valuable support during the research work. The laboratory facilities and analytical instrumentation provided at Manipur International University were instrumental in successfully conducting this research work.

    CONFLICT OF INTEREST

    The authors declare that no conflict of interest was reported regarding the publication of this article. 

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