Formaldehyde is a prominent volatile organic compound and also considered an indoor air pollutant. Chlorophytum comosum, an indoor plant, has been reported to metabolize indoor formaldehyde. But the phytotoxic effects of formaldehyde, being a pollutant, on C. comosum is not well explored. Furthermore, C. comocum responses that can be considered as markers at the physiological and biochemical level against formaldehyde stress are not yet investigated. Therefore, the current research study was aimed to evaluate such potential markers against formaldehyde in C. comosum. Briefly, C. comosum was exposed to 5, 10, and 20 ppm formaldehyde doses in an airtight glass chamber. Plant samples were then taken to analyze morpho-anatomical, physiological, and biochemical responses after short (2, 4, and 6 hours), medium (12 and 24 hours) and extended durations (48 and 96 hours) for each tested dose. Two-ways ANOVA followed by Tukey’s test at p ≤ 0.05 indicated that application of 10 and 20 ppm formaldehyde doses led to a significant incline in enzymatic antioxidants like CAT, GPX and SOD, and non-enzymatic parameters including TPC, TFC, T-AOC, carotenoids and intercellular CO2. However, formaldehyde application negatively affected the physiological responses of C. comosum by reducing its photosynthetic rate, transpiration rate and stomatal conductance. Additionally, extended exposure of C. comosum to 10 and 20 ppm formaldehyde doses led to visible leaf damage. Principal Component Analysis indicated that enzymatic (SOD, CAT and GPX) and non-enzymatic (MDA, TPC, TFC, TAO, carotenoids, TSS and intercellular CO2) parameters contributed the most to the total variance. Thus, these parameters have potential to serve as physiological and biochemical markers in C. comosum against formaldehyde stress.