The use of phthalates as plasticizers has been omnipresent, especially in cosmetics and food packaging, despite the proven effects on some organs of humans and animals. Therefore, alterations in living organisms due to phthalate exposure attract the attention of many scientists. Here, we demonstrate a mechanical and chemical investigation of the mentioned effects of di(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) on rat liver by utilizing scanning acoustic microscopy (SAM), Raman spectroscopy (RS) and inductively coupled plasma optical emission spectrometry (ICP-OES) for the first time in the literature, as far as we know. The combined analysis gives insights into the degree of modification in the tissue components and which chemicals lead to these modifications. Our study shows that the acoustic impedance values of tissues of DEHP and DBP delivered mother rats are higher than those of tissues of the control mother rat, while the acoustic impedance values of tissues of offspring rats of DEHP and DBP delivered mother rats do not differ significantly from those of tissues of the control offspring rats of the control mother rat. Besides, RS analysis shows how the incorporation of DEHP into liver tissues changes the configuration and conformation of lipids and fatty acids. ICP-OES results show increased element levels within the tissues of DEHP and DBP delivered rats. Therefore, we can say that phthalates cause modifications within the liver. This study is a preliminary effort to investigate tissues with a mechano-chemical probe.