A novel bis-Schiff base 4 has been synthesized in this manuscript, it is well characterised by different spectroscopic technique such as FT-IR, 1H &13C NMR spectroscopy and Mass spectrometry, TGA analysis has done to check the thermal stability of compound 4 and it complexes with Al(III) and citric acid. Under the UV–visible spectrophotometer, chemosensor 4 detects Al(III) and citric acid and then comprehensive analysis of real sample using aluminium foil and lemon juice solution toward compound 4 were carried out to enhance to support for sensing application. The limit of detection (LOD) and association constant (Ka) of synthesized chemosensor 4 towards Al(III) are found to be 9 × 10−6 M and 7 × 104 M whereas with citric acid are 2.6 × 10−6 M and 1.8 × 105 M respectively. The binding ratio of the compound 4 with Al(III) and citric acid are found to be 1:1 ratio. Anti-cancer activity of the chemosensor 4 has responded maximum reduction of cell viability up to 61.5 % to compare with untreated control cell, furthermore to assess anticancer activity computationally, molecular docking study of the compound 4 against protein 5hu9 has shown maximum binding energy (−11.41 kcal/mole).