The analytical detection and quantification of abuse drugs such as morphine (MOR) in biological samples are vital missions and remains to attract challenges for forensic toxicology, law enforcement, world antidoping organization, and social health fields. MOR, a benchmark analgesic drug known as "pain killer", is one of the powerful opioid medications for relieving pain, and overdose of MOR is toxic. In this article, novel promising chromium metal−organic framework nanoparticles [Cr(III)-MOF-NPs] were produced via facile synthesis and characterized using high-resolution transmission electron microscopy, field-emission scanning electron microscopy/energy-dispersive X-ray spectroscopy, mass spectrometry, X-ray photoelectron spectroscopy, elemental analysis, UV−vis, Fourier transform infrared, and thermogravimetry/differential scanning calorimetry, as well as photoluminescence (PL) investigation and magnetic properties. The PL study results revealed that the Cr(III)-MOF-NPs exhibited an emission band at 593 nm. The Cr(III)-MOF-NPs could be used in fast, selective, and sensitive MOR detection and quantification. Under the optimum experimental conditions, with the addition of MOR, a blueshift from 593 to 566 nm occurred with a remarkable PL intensity enhancement, and the color changed from brown to yellow (visually/naked-eye detection). The Cr(III)-MOF-NPs optical chemosensor exhibited a stable response for MOR in a concentration range between 0.1 and 350 nM. The detection and quantification limits were 0.167 and 0.443 nM, respectively, with a correlation coefficient (r 2 ) of 0.96. The developed PL chemosensor showed high selectivity for MOR over other competing interfering matrices. Moreover, the ultrasensitive chemosensor was extensively used for the determination of MOR spiked in different real samples (serum and urine samples) with acceptable recoveries and satisfactory results.