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N-carbamoylglutamate (NCG) is a feed additives obtained synthetically from sodium glutamate, which is a structural analogue of N-acetylglutamate. Current study is aimed at developing ion chromatographic method for the determination of NCG in rumen fluid. Successful determination was performed on cation-exchange column and non-suppressed conductivity detection with total rune time of 11 minutes. Simple one-step cleanup using anion exchange cartridge was used to remove interfering compounds. Method was applied to determine NCG in rumen fluid, which was obtained from three rumen fistulated dairy cows. Incubated NCG sample in rumen fluid was assessed at various time intervals (0, 2, 4, 6, 12 and 24 h at 39 ºC). Results showed that NCG was disappeared at the rate of 7.8, 12.8, 13.3, 14.2, and 17.8 % at 2, 4, 6, 12 and 24 h, respectively. It is concluded from the results that the procedure is quick and uncomplicated. NCG can be detected in rumen fluid without lengthy separation using cation-exchange chromatography. This technique can be applied for determination of NCG in rumen fluid as well as for practical use in animals feed.