Surface-enhanced laser desorption/ionization (SELDI) is a versatile and high-throughput mass spectrometry (MS) technique that uses a probe for extraction, enrichment, desorption, and ionization of target analytes. Here we report ordered mesoporous carbon as a new SELDI probe for rapid screening and identification of trace amount of toxic chemicals in a single drop of human whole blood without complicated sample preparation procedures. We demonstrate that ordered mesoporous carbon not only can selectively enrich a wide variety of low-mass toxic compounds from whole blood samples but also can be used as an excellent matrix to assist the laser desorption/ionization process of small molecules with low background noise, high repeatability, and good salt tolerance. High sensitivity (detection limits at ppt levels) and good reproducibility for typical toxic compounds were obtained. With CMK-8 as a SELDI probe, we successfully identified and screened six perfluorinated compounds in a single drop of whole blood collected from workers in a perfluorochemical plant. The method was also validated with complex samples such as human urine and environmental water samples. With distinct advantages such as simplicity, rapidness, minimal sample requirement, and high reliability, this method keeps great promise for various aspects of application.