Adapted MS/MS^ALL Shotgun Lipidomics Approach for Analysis of Cardiolipin Molecular Species

Cardiolipin (Ptd₂ Gro) is a complex, doubly charged phospholipid located in the inner mitochondrial membrane where it plays an essential role in regulating bioenergetics. Abnormalities in Ptd₂ Gro content or composition have been associated with mitochondrial dysfunction in a variety of disease states. Here, we report the development of an adapted high-resolution data-independent acquisition (DIA) MS/MS^ALL shotgun lipidomic method to enhance the accuracy and reproducibility of Ptd₂ Gro molecular species quantitation from biological samples. Utilizing the doubly charged molecular ions and the isotopic pattern with negative mode electrospray ionization mass spectrometry (ESI-MS) using an adapted MS/MS^ALL approach, we profiled more than 150 individual Ptd₂ Gro species, including monolysocardiolipin (MLPtd₂ Gro). The method described in this study demonstrated high reproducibility, sensitivity, and throughput with a wide dynamic range. This high-resolution MS/MS^ALL shotgun lipidomics approach could be extended to screening aberrations of Ptd₂ Gro metabolism involved in mitochondrial dysfunction in various pathological conditions and diseases.