Increasing amounts of ACV (Selleckchem, USA) dissolved in DMSO was suspended in PBS to make a concentration gradient. NanoDrop UV Spectrophotometer was used to determine the absorption peak of ACV at 252 nm. Typically, 2 μl of the desired solution was dropped onto the analysis port of the NanoDrop before closing the lid and starting the measurement. Each measurement was conducted thrice with three individual drops, and three separate replicates were used to determine the absorbance of every ACV concentration. The readings were noted down manually and entered into GraphPad Prism software to prepare the concentration versus absorbance curve.

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