Purpose: Our institution, the demand for [¹³N]ammonia is increasing, so we will introduce the manufacturing process, quality testing, and other processes for [¹³N]ammonia injection until it is supplied to patients. Materials and Methods: [¹³N]ammonia is produced by irradiating water containing 5 mmol of ethanol as a target material in a cyclotron manufactured by IBA at about 90 mA for 15 minutes. The quality of the prepared [¹³N]ammonia injection is tested according to USP standards. Results: The [¹³N]ammonia injection solution produced in a cyclotron passes through a cartridge and a sterile filter, and the final amount is about 200 mCi on average in about 2 mL volume, and the production time is about 20 minutes. All quality test results met USP standards. Conclusion: [¹³N]ammonia injection is a drug whose demand is gradually increasing due to its simple manufacturing process and manufacturing time that does not require synthesis equipment, relatively uncomplicated quality testing, and good visual image.