Background <p>The standardization and safety evaluation of traditional herbal formulations are critical prerequisites for their scientific validation and clinical translation, as emphasized by the World Health Organization. Chronic kidney disease, affecting more than 800&#xa0;million individuals globally, represents a major public health challenge with limited therapeutic options. Although several polyherbal formulations are described in traditional medicinal systems for renal disorders, systematic quality control and safety profiling remain largely inadequate. The present study aimed to comprehensively standardize and assess the safety of a classical Unani PHF, <i>Safoof Karafs</i>, traditionally used for renal ailments, in accordance with WHO and API guidelines.</p> Methods <p>The PHF was prepared using authenticated plant materials as per classical Unani texts. Standardization encompassed detailed physicochemical characterization and phytochemical profiling. Safety evaluation focused on the assessment of heavy metals, microbial load, aflatoxins and pesticide residues employing validated analytical techniques such as ICP-MS, LC-MS/MS, and GC-MS/MS. All analyses were performed in triplicate, and data were expressed as mean ± SEM.</p> Results <p>The PHF met all established physicochemical and phytochemical quality parameters and showed full compliance with WHO and API guidelines. Safety analysis demonstrated that heavy metals, aflatoxins, pesticide residues, microbial load, and pathogenic contaminants were within permissible limits.</p> Conclusion <p>This study establishes a comprehensive standardization and safety profile of <i>Safoof Karafs</i>, validating its quality, purity, and suitability for further pharmacological, preclinical, and clinical investigations. The findings support the integration of traditional PHF into evidence-based healthcare and align with WHO strategies for the global acceptance of traditional medicines.</p> Graphical abstract <p></p>