Identification and Quantitation of Signature Constituents of Dant‐Kanti, a Herbo‐Dental Formulation Using UHPLC, HPTLC, and GC–MS, and its Anti‐Inflammatory Effects on <i>Porphyromonas gingivalis</i> Lipopolysaccharide‐Induced Human Macrophages

ABSTRACT A herbo‐dental formulation, Dant‐Kanti natural toothpaste consists of 13 potent medicinal herbs. The present study aims to comprehensively identify the signature phytometabolites present in Dant‐Kanti natural toothpaste using high‐performance thin layer chromatography (HPTLC), ultra‐high‐performance liquid chromatography (UHPLC), and gas chromatography–mass spectrometry (GC–MS) methods. Among the identified phytometabolites, gallic acid, piperine, and anethole, were quantified by UHPLC, whereas camphor, menthol, thymol, and eugenol were quantified using GC–MS. An inductively coupled plasma‐mass spectrometry was used to estimate the mineral contents in Dant‐Kanti natural toothpaste. To the best of our knowledge, these studies are the first‐ever to identify all the signature components in an herbal dental formulation. These developed and validated methodologies were then used across 25 distinct batches of Dant‐Kanti natural toothpaste, in oder to ascertain the consistency and homogeneity. Both the UHPLC, and GC–MS methods used in this study were validated as per ICH Q2 (R1) guidelines, and were found to be suitable, linear ( r &gt; 0.99), and precise. In the cell‐biological study, Dant‐Kanti natural toothpaste showed a concentration‐dependent reduction in proinflammatory cytokine release, in human macrophages induced with Porphyromonas gingivalis lipopolysaccharide. This robust study methodology, validation, and results provide valuable insights into the complex composition of Dant‐Kanti natural toothpaste, and contribute to the understanding of its potential oral health benefits.