Protoporphyrin IX plasma and blood pharmacokinetics and brain tumor distribution determined by a validated LC-MS/MS method

Authors

William Knight, Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, 350 West Thomas Road, NRC 403, Phoenix, AZ, 85013-4496, USA.
Tigran Margaryan, Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, 350 West Thomas Road, NRC 403, Phoenix, AZ, 85013-4496, USA.
Kamal Shaik, Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, 350 West Thomas Road, NRC 403, Phoenix, AZ, 85013-4496, USA.
Nader Sanai, Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, 350 West Thomas Road, NRC 403, Phoenix, AZ, 85013-4496, USA.
Artak Tovmasyan, Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph's Hospital & Medical Center, 350 West Thomas Road, NRC 403, Phoenix, AZ, 85013-4496, USA. artak.tovmasyan@barrowneuro.org.

Document Type

Article

Abstract

Protoporphyrin IX (PPIX) is a fluorescent metabolite in the heme biosynthesis pathway, and cancer cells accumulate it when 5-aminolevulinic acid (5-ALA), a precursor, is administered. In the U.S., 5-ALA is approved for visualizing high-grade gliomas (HGG) during fluorescence-guided surgery. PPIX is also central to experimental photodynamic and sonodynamic therapies for HGG. Additionally, PPIX measurement is critical for diagnosing and monitoring porphyrias. Despite the need for a sensitive and rapid bioanalytical method for accurate quantification of PPIX in biospecimens, no reliable validation of an LC-MS/MS method is available due to challenges related to its chemical instability, poor solubility, and tendency to aggregate. This work is the first to present a fully validated, sensitive, and rapid LC-MS/MS method for determining PPIX levels in human plasma, blood, and brain tumors. The method overcomes stability concerns, achieving a 3.5-min total run-time with a concentration range of 1-2000 nmol/L for plasma and tumors, and 10-2000 nmol/L for blood. Application of the method in a clinical trial, which assesses sonodynamic therapy for HGG patients, shows significant PPIX production, peaking in plasma and blood six hours post-5-ALA administration. In recurrent HGG patients, PPIX levels were notably higher in gadolinium-enhancing tumor regions compared to non-enhancing areas, indicating preferential accumulation in tumors.

Keywords

5-aminolevulinic acid (5-ALA), Brain tumor, Liquid chromatography tandem mass spectrometry (LC–MS/MS), Photodynamic therapy, Protoporphyrin IX (PPIX), Sonodynamic therapy

Medical Subject Headings

Protoporphyrins (blood, pharmacokinetics); Humans; Tandem Mass Spectrometry (methods); Brain Neoplasms (blood, metabolism, pathology); Chromatography, Liquid (methods); Aminolevulinic Acid; Glioma (blood, metabolism); Photosensitizing Agents (pharmacokinetics); Male; Female; Liquid Chromatography-Mass Spectrometry

Publication Date

7-1-2025

Publication Title

Scientific reports

E-ISSN

2045-2322

Volume

15

Issue

1

First Page

21521

PubMed ID

40595976

Digital Object Identifier (DOI)

10.1038/s41598-025-05780-w

Recommended Citation

Knight, William; Margaryan, Tigran; Shaik, Kamal; Sanai, Nader; and Tovmasyan, Artak, "Protoporphyrin IX plasma and blood pharmacokinetics and brain tumor distribution determined by a validated LC-MS/MS method" (2025). Translational Neuroscience. 2480.
https://scholar.barrowneuro.org/neurobiology/2480