Direct proof of use
The clearest direct-use incident is the 8 August 2014 attack on the Ukrainian border service department in Milove, Luhansk province. International Partnership for Human Rights and Truth Hounds reported that the facility was attacked with RPO Shmel flamethrowers, SPG grenades, and GP-25 grenades, and that their field mission found several GP-25 hits on the garage of the border department facility.
The Milove report treated the attack as part of a set of cross-border incidents in Luhansk province and attributed the documented attacks to people operating from or returning to Russian territory. The GP-25 evidence in that report is therefore evidence of grenade-launcher use in a specific border-facility attack, not a complete accounting of every GP-25 in the theater.
Sources: Investigation of Cross-Border Attacks in Eastern Ukraine
Recovered and traced launchers
Conflict Armament Research separately documented two 40 mm GP-25 under-barrel grenade launchers in Mariupol on 18 September 2019 during its investigation of weapons recovered from the armed formations of the self-declared Donetsk and Luhansk People's Republics. CAR determined that the two launchers were manufactured on Russian territory.
Ukrainian trace responses to CAR said the two GP-25 launchers with serial numbers 15 4657 and 52 6734 had not been in service with the Armed Forces of Ukraine, had not been recorded as stolen, lost, or written off, and had not been transferred to other military units. That evidence supports recovered conflict materiel and Russian-side provenance, while the report does not by itself identify a firing incident for those two recovered launchers.
Sources: Weapons of the War in Ukraine
Ukrainian-side training
Ukrainian forces also had GP-25s in the conflict-era force structure. DVIDS documented soldiers from the 1st Airmobile Battalion, 79th Air Assault Brigade training with GP-25 grenade launchers at the Yavoriv Combat Training Center on 3 May 2017.
The DVIDS caption says the soldiers were being taught to employ the grenade launchers during defensive operations during the battalion's rotation at the training center. This is training and fielding evidence for the Ukrainian side, distinct from the Milove firing incident and the CAR recovery evidence.
Sources: Ukrainian GP-25 Grenade Launcher Training