Chemical Fingerprinting of Spiranthes spiralis L. Methanol Seed Extract: Spectroscopic, Chromatographic, and Computational Approaches

TL;DR

This study identifies the chemical composition and bioactive potential of Spiranthes spiralis seeds using various analytical and computational methods.

Contribution

The study provides a comprehensive chemical fingerprint and pharmacological evaluation of S. spiralis seeds using integrated experimental and computational approaches.

Findings

S. spiralis seeds contain high phenolic content and strong antioxidant activity with an IC50 of 0.21 mg/mL.

GC–MS identified 20–22 bioactive compounds, including 2,2-dimethoxybutane and hydrazinecarbothioamide, with high binding affinity to GPR52.

Computational analyses revealed varying reactivity and toxicity profiles among the identified compounds.

Abstract

In this study, the morphological properties, antioxidant activities, and phytochemical content of Spiranthes spiralis L. seed methanol extracts were characterized and analyzed in silico. Microscopic analysis revealed a fusiform seed shape characterized by prominent basal cells with thick, slanted ridges and polygonal testa structures. Fourier transform infrared spectroscopy (FTIR) identified distinct absorption bands corresponding to O–H, –CH2–, C=O, and other functional groups, indicating the presence of phenolic compounds, proteins, and polysaccharides. The total phenolic content was measured at 24.65 ± 1.43 mg GAE/g dry weight, while flavonoid and tannin contents were determined to be 43.98 ± 3.10 mg QE/g dry weight and 2.13 ± 0.13 mg GAE/g dry weight, respectively. Antioxidant activity, assessed via DPPH radical scavenging, yielded an IC50 value of 0.21 mg/mL, indicating a strong antioxidant potential associated with the phenolic content. The gas chromatography–mass spectrometry (GC–MS) analysis revealed 20 bioactive compounds, with 2,2‐dimethoxybutane and hydrazinecarbothioamide among the major constituents. Molecular docking indicated high binding affinities to the GPR52 receptor, with 4,4,6,6‐tetramethyl‐1,3‐dioxane displaying the lowest binding energy (−6.3 kcal/mol). Absorption, distribution, metabolism, excretion, and toxicity (ADMET) predictions revealed mixed toxicity profiles, as hydroxyacetic acid, hydrazide, and hydrazinecarbothioamide were flagged for potential mutagenicity. Further computational analysis, including Molecular Electrostatic Potential (MEP) and non‐covalent interaction (NCI) and reduced density gradient (RDG) mapping, supported the presence of well‐defined electrostatic regions and weak interaction zones. Additionally, the highest occupied molecular orbital (HOMO)–lowest unoccupied molecular orbital (LUMO) and global reactivity descriptors suggested that hydrazinecarbothioamide exhibits high electrophilicity and reactivity. At the same time, 2,2‐dimethoxybutane was found to be more chemically stable and less reactive. Overall, the findings emphasize the phytochemical richness and bioactive potential of
S. spiralis
seeds, offering promising perspectives for future pharmacological and biotechnological applications.

Spiranthes spiralis L. seed methanol extract was morphologically and chemically characterized using microscopy, FT‐IR, GC–MS, and in silico analyses. The extract exhibited high phenolic content and strong DPPH radical scavenging activity, while GC–MS identified 22 bioactive compounds with notable binding affinity toward the GPR52 receptor. Molecular docking, ADMET, and electronic structure analyses highlighted the bioactive and pharmacological potential of
S. spiralis
seeds.

Figures