<p>The potential of pollen from Indian bees as a practical therapeutic agent is still largely unknown. It is crucial to carefully assess, evaluate, and set national standards for the quality of Indian bee pollen because of the variations in chemical composition across different geographic regions. The chemical composition of bee pollen samples varies significantly between countries, and there are no common criteria for assessing the qualities of bee pollen. Therefore, the present study aimed to develop and validate a method for the simultaneous estimation of rutin, rosmarinic acid, kaempferol, and caffeic acid in Indian coconut bee pollen with the help of thin-layer chromatography–densitometric technique. A mixture of toluene–ethyl acetate–methanol–formic acid–water (3:6:1:0.5:0.5, <i>V/V</i>) was used as the mobile phase to accomplish good chromatographic separation using an ultraviolet–visible (UV–Vis) detector at wavelengths of 366 and 328&#xa0;nm. The retardation factors for rutin, rosmarinic acid, caffeic acid, and kaempferol were set as 0.090, 0.553, 0.610, and 0.683, respectively. A good linear relationship was achieved for rutin, rosmarinic acid, caffeic acid, and kaempferol in the linear regression data for the calibration curves over the concentration ranges of 400–900 (<i>R</i>² = 0.9979), 1000–4500 (<i>R</i>² = 0.9995) 500–3000 (<i>R</i>² = 0.9909), and 1500–4000&#xa0;ng/band (<i>R</i>² = 0.9979), respectively. The established method was found to be exact, accurate, and resilient. Following the method’s application to a commercial formulation (Natural Bee Pollen Capsules), the percentage amounts of rosmarinic acid and kaempferol were determined to be 4.13 ± 0.0001 and 0.205 ± 0.001, respectively.</p>