Introduction: Peptic ulcer is a disease which is unrelated to non-steroidal anti-inflammatory drugs (NSAIDs) or Helicobacter pylori infection. The higher risk of bleeding and ulcer recurrence highlights the clinical importance of analyzing the changing trends of peptic ulcer disease in developing countries. Aims: To assess the proportion of non-NSAID, non-H. pylori peptic ulcer disease in a Pakistani cohort of patients with peptic ulcer disease managed at Mayo Hospital Lahore; and to compare the gastric and duodenal ulcer subgroups in these patients. Methods: The methods that we have used needed patients who had been diagnosed with peptic ulcer disease to be screened for a history of NSAID use and after the screening of those with a negative history were tested for H. pylori through combined tests of rapid urease and 14C-urea breath test (UBT). Only the cases which showed negative results for these two tests were considered ‘H. pylori-negative’. Serum gastrin was measured in all patients included in the study. Results: 74 gastric ulcer (GU) and fifty-four duodenal ulcer (DU) patients were included in the study. Of these, thirty-six patients GU (45.9%) and sixteen patients DU (29.6%) were H. pylori-negative. The proportion of non- NSAID and non-H. pylori gastric ulcers was significantly higher than duodenal ulcers (p <0.05). However, patients who tested negative for H. pylori did not differ significantly from those who tested positive with regard to age, gender, serum gastrin level, and presence of risk factors, like smoking and alcoholism. Conclusion: The current study indicates existence of high proportion of non-NSAID, non-H. pylori peptic ulcer disease in Pakistani patients.