Background interferences are perhaps an unavoidable part of analytical detection schemes, particularly when analyzing trace level samples or when using detectors with high levels of sensitivity. In liquid chromatography, mobile phase solvents are routinely filtered using membrane filter disks to trap particulates in hopes of minimizing contamination, providing improvements in data output and instrumental operation. In this study, we report that one such filter disk leads to a significant level of contamination in LC and LC/MS experiments. Extractable compounds from nylon membrane filters generate significant background signals in UV absorption chromatograms at 214 nm, and are also detected by electrospray ionization mass spectrometry, with nominal m/z values of 453 and 679. It is shown that rinsing the nylon membranes before their use can reduce, but will not eliminate, the extractable contaminants from the mobile phase. Through MS and tandem MS analysis, we have identified these contaminants as cyclic oligomers of polyamide 66. Based on these results, it is recommended that nylon membrane filters be avoided when conducting trace level analysis, particularly when conducting LC/MS experiments. (J Am Soc Mass Spectrom 2006, 17, 652-656)