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Background and objectives: Laboratory medicine is equally challenged by economic and new technological pressures. Clinical laboratories have undergone major change due to advancement of technology, which has improved the decision making of clinicians but introduced the risk of errors. This study aimed to evaluate the errors that occurred in the pre-analytical phase of laboratory testing. Methods: This was a prospective observational study that was done in the Clinical Biochemistry Laboratory of a tertiary care center from June 2016 to May 2017. The path of the sample was analyzed from sample collection to transport. Frequency of deficiencies in the request forms and different types of pre-analytical errors were recorded. Results: During the study period, the frequency of pre-analytical errors was about 3.1%. Sample hemolysis was the predominant error in sample collected from both indoor and outdoor patients. Conclusion: Proper management of pre-analytical errors requires continuous evaluation of source of errors, taking corrective measures, and significant interdepartmental cooperation.