Efficient and cost effective measurement of the refractive index profile in an optical fiber is a significant technical job to design and manufacture in-fiber photonic devices and communication systems. For instance, to design fiber gratings, it is required to estimate the refractive index modulation to be inscribed by the fabrication apparatus such as ultraviolet or infrared lasers. Mach-Zehnder interferometer (MZI) based quantification of refractive index change written in single mode microfiber by femtosecond laser radiation is presented in this study. The MZI is constructed by splicing a microfiber (core diameter: 3.75 μm, cladding diameter: 40 μm) between standard single mode fibers. To measure the RI inscribed by infrared femtosecond laser, 200 μm length of the core within the MZI was scanned with laser radiation. As the higher index was written within 200 μm length of the core, the transmission spectrum of the interferometer displayed a corresponding red shift. The observed spectral shift was used to calculate the amount of refractive index change inscribed by the femtosecond irradiation. For the MZI length of 3.25 mm, and spectral shift of 0.8 nm, the calculated refractive index was found to be 0.00022. The reported results display excellent agreement between theory and experimental findings. Demonstrated method provides simple yet very effective on-site measurement of index change in optical fibers. Since the MZI can be constructed in diverse fiber types, this technique offers flexibility to quantify index change in various optical fibers.