Comprehensive contaminant screening and microbial safety evaluation of a classical Unani anti-arthritic formulation

Safety assurance and chemical profiling of traditional herbal formulations are critical due to global reports of toxic contamination in raw materials. This analytical study aimed to comprehensively evaluate the toxicological and microbial safety profile of a classical Unani Pharmacopoeial Formulation (UPF), traditionally used in the management of osteoarthritis, by rigorously applying advanced analytical techniques aligned with modern quality control standards. The formulation, prepared from detoxified crude ingredients in pharmacopoeial proportions, was subjected to a comprehensive chemical safety screening. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was employed for the precise quantification of four priority toxic heavy metals (As, Pb, Cd, Hg). Aflatoxins (AFB₁, AFB₂, AFG₁, AFG₂) were analyzed using highly sensitive Liquid Chromatography-Tandem Mass Spectrometry (LC–MS/MS). Furthermore, 34 target pesticide residues were screened and quantified via Gas Chromatography-Tandem Mass Spectrometry (GC–MS/MS) under optimized Multiple Reaction Monitoring (MRM) conditions. A final microbial evaluation included Total Aerobic Microbial Count and targeted pathogen detection. The analytical results confirmed the high purity of the final formulation. All four toxic heavy metals (As, Pb, Cd, Hg) were found to be below the Limit of Quantification (LOQ) or well within the permissible limits set by WHO/API guidelines. Similarly, trace levels of aflatoxins were detected but remained significantly below the established thresholds. Critically, all 34 screened pesticide residues were found to be below the LOQ. Furthermore, the microbial assessment demonstrated that the Total Aerobic Bacterial and Fungal counts were within acceptable limits, with no targeted pathogens identified. The findings conclusively establish that the Unani Pharmacopoeial Formulation, prepared using detoxified ingredients, meets stringent contemporary chemical and microbial safety standards. This comprehensive toxicological profile validates the quality of the preparation, providing the essential chemical evidence required to support its safe transition for further pharmacological and rigorous clinical development in musculoskeletal disorders.