By in vitro fertilization, oocytes can be removed and the embryo can be cultured, and then trans cervically replaced when they reach cleavage or at the blastocyst stage. The characterization of the follicular fluid is important for the treatment process. Women who applied to the Academic Hospital in vitro fertilization (IVF) Center diagnosed with idiopathic female infertility (IFI) were sought in the patient group. Demographics and clinical gonadotropin measurements of the study population were recorded. Of the 116 follicular fluid samples (n=58 male-induced infertility; n=58 control) were analyzed using the FTIR system. To identify FTIR spectral characteristics of follicular fluids associated with an ovarian reserve and reproductive hormone levels from control and IFI, six machine learning methods and multivariate analysis were used. To assess the quantitative information about the total biochemical composition of a follicular fluid across various diagnoses. FTIR spectra showed a higher level of vibrations corresponding to lipids and a lower level of amide vibrations in the IFI group. Furthermore, the T square plot from Partial Last Square (PLS) analysis showed, that these vibrations can be used to distinguish IFI from the control group which was obtained by principal component analysis (PCA). Proteins and lipids play an important role in the development of IFI. The absorption dynamics of FTIR spectra showed wavenumbers with around 100% discrimination probability, which means, that the presented wavenumbers can be used as a spectroscopic marker of IFI. Also, six machine learning methods showed, that classification accuracy for the original set was from 93.75% to 100% depending on the learning algorithm used. These results can inform about IFI women's follicular fluid has biomacromolecular differentiation in their follicular fluid. By using a safe and effective tool for the characterization of changes in follicular fluid during in vitro fertilization, this study builds upon a comprehensive examination of the idiopathic female infertility remodeling process in human studies. We anticipate that this technology will be a valuable adjunct for clinical studies.