An anaphylactic reaction during general anesthesia may cause fatal complications. Perioperative anaphylaxis can occur in response to different medications. To our knowledge, pyridostigmine or combination of pyridostigmine with glycopyrrolate-induced anaphylactic reaction during general anesthesia has not been reported. We report a case of a suspected anaphylactic reaction to these drugs. To reduce the risk of complicated perioperative anaphylactic events, prompt assessment and treatment are needed. Moreover, any undesirable exposure to confirmed allergens should be avoided as part of secondary prevention.