Acute pancreatitis is a rather common abdominal disorder. In most patients the disease is mild, but about 20% of cases develop a severe necrotizing form of the disease with complications. In an emergency setting, the diagnosis of acute pancreatitis remains problematic and several patients with severe disease are diagnosed only at autopsy. Measurements of amylase or lipase are the principal laboratory methods for diagnosing acute pancreatitis. However, their sensitivity and specificity are generally considered unsatisfactory. Recent advances in the knowledge of the pathogenesis of acute pancreatitis and advances in laboratory technology have revealed new diagnostic possibilities. Especially assays based on trypsin pathophysiology have brought new alternatives for diagnostics and severity grading of the disease. Additionally, development of phospholipase A2 determinations and discovery of a new pancreatic protein, pancreatitis-associated protein, are very interesting. This article summarizes the value of new methods in the laboratory diagnostics of acute pancreatitis.