The aim of the study to investigate the Physicochemical analysis, Fluorescence analysis, Extraction, Phytochemical screening, Thin-layer chromatography (TLC) and High Performance thin-layer chromatography (HPTLC) of leaves of Amaranthus cruentus (A. cruentus). Physicochemical analysis, including the determination of various parameters such as loss on drying, swelling index, foaming index, total ash, acid-insoluble ash, water-soluble ash, extractive value, and fluorescence analysis was conducted on the powdered leaves. To extract the bioactive compounds, a Continuous hot Soxhlet apparatus was employed using solvents with increasing polarity, namely petroleum ether, chloroform, and ethyl acetate. Phytochemical screening was then performed on these extracts to identify the presence of various phytoconstituents. Thinlayer chromatography (TLC) and High-performance thin layer chromatography (HPTLC) were carried out exclusively on the ethyl acetate extract. The solvent system used for TLC and HPTLC was a mixture of toluene and ethyl acetate in a ratio of 7:3. HPTLC was performed with two marker compounds, Gallic acid, and Rutin, as these were the predominant phytoconstituents identified in the ethyl acetate extract. For the quantitative HPTLC analysis, a precoated silica gel 60 F254 plate was utilized, and UV detection was conducted at a wavelength of 254nm. This allowed for the precise determination of the amounts of Gallic acid and Rutin present in the ethyl acetate extract. The Physicochemical evaluation of powder indicated value of loss on drying was found 5.1%, swelling index value was 0.23%, the foaming index less than 100, total ash value was 11.86%, acid-insoluble ash value was 2.56%, water soluble ash value was 3.33% and pet. ether extractive value was 1.45%, chloroform extractive value was 1.21% and ethyl-acetate extractive value was 1.36%. In UV and visible light, many fluorescent colors were seen during fluorescence analysis. Pet. ether extraction value was 3.04%, chloroform extraction value was 3.41% and ethyl acetate extraction value was 3.64%. The preliminary phytochemical analysis of various extracts revealed the presence of alkaloids, carbohydrates, glycosides, amino acids, proteins, steroids, tannins and phenolics, saponins, flavonoids, fats, and oils. Additionally, TLC was performed on the ethyl acetate extract, resulting in the identification of seven unknown phytoconstituents. To further investigate the presence of specific compounds, HPTLC technique was employed. This technique enabled the detection and quantification of gallic acid and rutin. The Rf (retention factor) values obtained for gallic acid and rutin were 0.05 and 0.88, respectively. The amounts of gallic acid and rutin were determined to be 958.69 µg/100 mg and 107.65 µg/100 mg, respectively. These findings provide valuable insights into the composition and quantity of these compounds in the analyzed extract. The present physicochemical parameters, fluorescence, extraction, phytochemical, TLC and HPTLC study results could help in standar...