Medicinal plants have long been used to cure a variety of infections (Tadese et al., 2022). Herbal medicines are prepared from the extract of the whole plant and from the leaves, roots, bark, seeds, and flowers. They can be used orally, inhaled, or directly on the skin (Bele et al., 2011). Medicinal plants contain a variety of molecules, many of which have antimicrobial and antioxidant effects, protecting the human body from infections as well as cellular oxidation. Consequently, it is important to describe the antioxidant and antimicrobial value of various medicinal plants (Juca et al., 2020).

The medicinal plant Solanum nigrum belongs to the Solanaceae family of plants. In most countries, they are semi-cultivated and mainly used as a vegetable and fruit supply. It is famous, as black nightshade is a weed that grows in damp conditions in a variety of soil types, including dry, rocky, shallow, and rich soil (Chen et al., 2022, Chen et al., 2022). Traditionally the use of Solanum nigrum is not limited to dysentery, skin eczema, swelling, prostatitis, and bronchitis. It has a wide range of antioxidant, immunomodulating, larvicidal, and antibacterial activities (Chen et al., 2022, Gao et al., 2021, Jaradat et al., 2016). The edible berries of Solanum nigrum are a rich source of bioactive compounds, which possess antioxidant and anti-inflammatory properties (Mani et al., 2022, Muthu et al., 2001). The antimicrobial activity of Solanum nigrum is attributed to the presence of bioactive compounds not limited to flavonoids, alkaloids, steroids, organic acids, and glycosides (Chen et al., 2022, Chen et al., 2022).

The global burden of bacterial skin diseases in 2021 was nearly 90 million, while it is expected to reach 1.2 billion in 2045. Bacterial skin infections are highly prevalent in developing countries like Pakistan (Aman et al., 2017, Gu et al., 2025). Staphylococcus aureus and Staphylococcus epidermidis are among the common bacteria causing skin infections (Del, 2020, Serra et al., 2015).

The recent emergence of multi-drug resistance (MDR) is alarming in S. aureus and S. epidermidis (Moglad and Altayb, 2022). Further, methicillin-resistant S. epidermidis (MRSE) and methicillin-resistant S. aureus (MRSA) are global challenges to public health. MRSA and MRSE harbour resistance to antibiotics due to MecA penicillin-binding protein 2 (MecA-PBP2), a 668 amino acid product of the mecA gene that has a transpeptidase activity and is involved in the peptidoglycan synthesis of bacterial cell wall (Mariana et al., 2001). Localised skin infections caused by resistant bacteria are difficult to treat using available antibiotics (Chen et al., 2022, Mozzillo et al., 2010, Parmanik et al., 2022). There is a global urgency for exploring novel antimicrobial sources to combat MRSA and MRSE. Further, toxicity, efficacy, and cost-effectiveness issues related to current antibiotics have driven research into medicinal plants as alternative sources of anti-infective compounds (Abdallah et al., 2023).

Liquid chromatography mass spectrometry has been successfully employed to identify bioactive compounds from plants (Tabassum et al., 2022, Juhar et al., 2025). Similarly, online tools have been significantly used to explore medicinal molecules from natural products. Lipinski’s rule, water solubility prediction, absorption, distribution, metabolism, excretion, and toxicity (ADMET), and molecular docking are useful for biological and synthetic chemistry, ligand interactions, drug-likeness, and toxicity of lead molecules (Bouamrane et al., 2022, Priyanka et al., 2024). The molecular docking and dynamics tools were previously used to explore antidiabetic, anticancer, and antibacterial compounds of Solanum nigrum and other medicinal plants (Belitibo et al., 2024, Chen et al., 2023, Sapkota et al., 2021). Furthermore, computational tools are vital to modern drug discovery and help to screen large compound libraries, ease the process, and optimise antibacterial compounds against MDR bacterial and fungal human pathogens (Mustapha et al., 2024, Shah et al., 2025).

From literature mining, few studies have reported plant activity against MRSA and MRSE (An et al., 2025; He et al., 2024; Janesha et al., 2020). Several studies are available on the antibacterial activity of Solanum nigrum extract (Belitibo et al., 2024, Chen et al., 2022); however, there is a lack of data on the Solanum nigrum high-performance liquid chromatography (HPLC) fractions against MRSA and MRSE.

The present study for the first time reported the HPLC fractions of Solanum nigrum ethanol extracts against MRSA and MRSE skin isolates. Bioactive fractions were further processed for compound identification and druggable lead molecules. Findings of the study will help to characterise the antibacterial compounds against MRSA and MRSE.