A sensitive optical sensor based on hollow core microstructure optical fibers modified with deep eutectic solvent was produced for the first time. An easy procedure for the modification of hollow-core microstructure optical fibers with deep eutectic solvent was developed. Deep eutectic solvents based on natural monoterpenoids and fatty acids were investigated for glass surface modification. The sensor was used for the determination of non-steroidal anti-inflammatory drugs (mefenamic acid, diclofenac, flurbiprofen and ketoprofen) in human urine samples. The mechanism of the sensor response was investigated and discussed. Liquid-phase microextraction of non-steroidal anti-inflammatory drugs was implemented in deep eutectic solvent phase supported in the inner surface of hollow-core microstructure optical fibers followed by transmission spectra measurement in one analytical device. The preconcentration step performed directly in the analytical device allowed to obtain high sensitivity and selectivity. The limits of detection calculated from the calibration plots based on 3σ were 3 μg L⁻¹ for all target analytes.