β-Lactam antibiotics are the most widely used class of drugs for the treatment of bacterial infections in human for many years. As antibiotic resistance can be an issue, rapid methods of analysis are needed. Here, a method for rapid LC-MS/MS quantification of 9 B-lactam antibiotics in human samples on the SCIEX QTRAP 4500 System was developed. The method uses a simple protein precipitation and fast chromatography and provides sensitivity, linearity, and robustness for such analysis in plasma samples.