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Poster Presentations
Day 2, June 23（Mon.）
Room P (Maesato East, Foyer, Ocean Wing)
- 2P-AM-22
Mass spectrometry based on a nitrilotriacetic acid affinity probe for the rapid enrichment and comprehensive profiling of cellular porphyrins
(1IoC Academia Sinica, 2FHS Macau Univ, 3DoC NTU)
oMei-Chun Tseng1, Elias Mernie1, Rofeamor Obena1, Fu-Lien Huang2, Tzu-Ming Liu2, Yu-Ju Chen1,3
Porphyrins are crucial macromolecules in heme biosynthesis, essential for oxygen transport, electron transfer, and nitric oxide synthesis. Disruptions in this pathway can lead to the accumulation of intermediate porphyrins, which serve as biomarkers for diseases such as porphyria and cancer. High-performance liquid chromatography (HPLC) with UV-visible and fluorescence detection is commonly used for porphyrin analysis. However, spectral overlap and interference from biological metabolites complicate accurate detection. To address this, we developed a novel analytical platform integrating immobilized metal affinity chromatography (IMAC) with magnetic nanoparticles (MNP). Using nitrilotriacetic acid (NTA)-conjugated MNP (NTA@MNP), we efficiently purified and enriched porphyrins with minimal sample loss. Fe³⁺-activated NTA@MNP enabled identification via matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Additionally, liquid chromatography-electrospray ionization-ion mobility tandem mass spectrometry (LC-MS/MS) facilitated the separation of porphyrin species in human lung cancer cell lines. This approach identified protoporphyrin IX (PPIX) and 12 putative endogenous porphyrin metabolites in PC9 cancer cells. Our method offers a robust tool for porphyrin metabolomics, enhancing the study of porphyrin-related diseases