There are many factors that contribute to the variation in detection probabilities of proteins in LC-MS/MS experiments, and currently little is known about their relative importance. In this study, we analyze the effect of competition for detection between coeluting peptides on peptide detection probability. Using a novel method for estimating peptide detection probabilities, we show that these probabilities can vary by an order of magnitude between peptides that elute from the liquid chromatograph at the same time as many other peptides and those that elute with fewer other peptides. To explore these results, we use a mathematical model to show that competition for detection between peptides is expected to be a major source of missed detections in complex mixtures because there will be many MS/MS scanning intervals that contain more coeluting peptides than can be subjected to MS/MS analysis. Our data and simulation results show that the number of coeluting peptides is a primary determinant of whether a peptide will be detected. In our data, this had a several-fold larger effect on peptide detection probability than did peptide abundance. Furthermore, the distribution of elution times for the most frequently detected peptides was strongly shifted toward values where there were few coeluting peptides, indicating that the number of coeluting peptides is a major determinant of whether a peptide is proteotypic.