Pancreatic infection is the most important risk factor in patients with necrotizing pancreatitis. It occurs in 40-70% of all patients suffering from the disease and influences their prognosis and clinical management. Whether antibiotic treatment can influence the infection rate and improve the patient’s prognosis is a subject of intense discussion. Little data on the concentrations of antibiotics in human pancreatic tissue are available, although they are regarded as the gold standard for evaluating antibiotic treatment protocols. We determined the concentrations of twelve broad-spectrum antibiotics and one β-lactamase inhibitor in human pancreatic tissue. All antibiotics were administered intravenously in patients undergoing pancreatic resection. Tissue concentrations were determined by high-pressure liquid chromatography. The potential clinical effectiveness in necrotizing pancreatitis of each antibiotic drug was evaluated by calculating an efficacy factor which took into account the frequency of bacteria found in the pancreatic infection, the measured pancreatic tissue levels and the minimal inhibitory concentrations (MIC’s) against the relevant bacteria. Among the tested antibiotics, imipenem, ciprofloxacin, ofloxacin, ceftriaxone, cefotaxime, ceftizoxime, cefotiam, piperacillin, mezlozillin, metronidazole and tazobactam were detected in pancreatic tissue at concentrations exceeding the MICs of most of the relevant bacteria. According to efficacy factor analysis, imipenem, ciprofloxacin and ofloxacin are the antibiotics which should be preferred for treatment of pancreatic infection. A first controlled clinical study with prophylactic imipenem in patients with severe acute pancreatitis showed a reduction in the rate of sepsis. This promising result needs further confirmation and emphasizes that the antibiotic regimen in necrotizing pancreatitis should be chosen according to the pharmacokinetic properties of the antibiotic drug in the human pancreas.