Analysis and detection of micronutrients is important for the reduction of the global burden of malnutrition-related disease. A relatively new technique, plasma pen atmospheric mass spectrometry (PPAMS) was applied in a comprehensive evaluation for rapid, simultaneous detection of the key micronutrients: zinc, iron, folate, vitamin A, and iodine. PPAMS was performed through the coupling of a low-temperature plasma pen to an atmospheric mass spectrometer (MS). The effectiveness of the PPAMS system was demonstrated through the generation of characteristic mass spectra and tandem mass spectra on neat micronutrient powders suspended on double sided tape. The analytical performance and ability to qualitatively separate out the nutrients from a complex biological solution and each other was then assessed through the application of PPAMS on a sample matrix of micronutrients in porcine plasma in which nutrient concentration is varied from high blood level concentrations (HBLCs) to low blood level concentrations (LBLCs). A multivariate software model, principal components analysis (PCA) was then used to qualitatively separate the fragments obtained by nutrient type. The resulting PCA scores plots of the positive ion spectra from each mixed sample showed excellent separation of HBLCs and LBLCs of single nutrients at the 95% confidence level.1 The associated PCA loadings plots showed that key loadings could be attributed to the expected micronutrient fragments. The PPAMS technique was successfully demonstrated and compared with traditional MS techniques: time-of-flight secondary ion mass spectrometry (ToF-SIMS) and electrospray ionization mass spectrometry (ESI-MS). Separation of the nutrients at concentrations relevant for human blood-based nutrient detection was possible in both ESI-MS and PPAMS. However, only PPAMS could detect the nutrients at physiological concentrations from porcine plasma. ToF-SIMS could detect the nutrients from plasma solution but required 5× to 1000× higher concentrations of folate, vitamin A, and iodine to achieve adequate separation of the micronutrients via PCA.