Background: Incidences of death and morbidity are high worldwide for acute neuroinflammatory disorders like encephalitis. The most helpful fluid for examining brain metabolism is cerebral spinal fluid, which also offers a crucial chance to identify neuro-inflammation in conditions affecting the human central nervous system. This study aimed to assess the role of nitric oxide as a cerebrospinal fluid metabolomics in diagnosis of neuroinflammatory diseases. Patients and methods: This case-control study was conducted in the neurology unit in the pediatric and clinical pathology departments of Zagazig University Hospital on 52 cases with neuroinflammatory conditions, they were equally divided into two groups; Control group included 26 patients (16 males and 10 females) with gray and white matter diseases and a case group of 26 patients (12 males and 14 females) with inflammatory neurological diseases. Cerebrospinal fluid were analysis by ELISA to assay the level of Nitric oxide in samples. Results: This study revealed that cerebrospinal fluid Nitric oxide levels, along with CSF Protein and CSF Cell Count, were significantly elevated in children with inflammatory neurological diseases compared to those with grey and white matter neurological conditions p<0.05. CSF Nitric oxide, with a cut-off level of ≥60.38 (mol/l), emerges as a highly sensitive (100%) and reasonably specific (88.5%) biomarker for discriminating neuroinflammatory diseases in children from other neurological conditions. Conclusion: Can be used CSF Nitric oxide as a valuable tool for diagnosing and monitoring neuroinflammatory disorders in pediatric patients, offering a potential avenue for early intervention and improved patient care