A method for the quantification of 8-methoxypsoralen by mass spectrometry for offline extracorporeal photopheresis

Background : Extracorporeal photopheresis (ECP) is an efficient method to treat various autoimmune diseases, cutaneous T-cell lymphoma, and graft- versus -host disease. It is based on the ex vivo inactivation of lymphocytes by 8-methoxypsoralen (8-MOP)/UV light treatment. Despite the adhesive, lipop...

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Published in:Photochemical & photobiological sciences 2017-02, Vol.16 (2), p.193-2
Main Authors: Hähnel, Viola, Dormann, Frauke, Nitsopoulos, Athanasios, Friedle, Albrecht, Ahrens, Norbert
Format: Article
Language:English
Subjects:
Adolescent
Adult
Biochemistry
Biomaterials
Blood Chemical Analysis - methods
Chemistry
Child
Chromatography, High Pressure Liquid
Female
Gas Chromatography-Mass Spectrometry
Humans
Lymphocytes - drug effects
Lymphocytes - metabolism
Lymphocytes - radiation effects
Male
Methoxsalen - analysis
Middle Aged
Photopheresis
Photosensitizing Agents - analysis
Physical Chemistry
Plant Sciences
Tandem Mass Spectrometry
Ultraviolet Rays
Young Adult
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Summary:Background : Extracorporeal photopheresis (ECP) is an efficient method to treat various autoimmune diseases, cutaneous T-cell lymphoma, and graft- versus -host disease. It is based on the ex vivo inactivation of lymphocytes by 8-methoxypsoralen (8-MOP)/UV light treatment. Despite the adhesive, lipophilic nature of 8-MOP, no quality control is established for the ECP procedure. Methods : We developed a sensitive high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) assay to monitor residual 8-MOP concentration after UVA irradiation in the whole blood supernatant after acetonitrile precipitation. Results : The preanalytical stability of 8-MOP exceeded 7 days, allowing batch mode analysis. Linearity was determined with R 2 above 0.99. The 8-MOP concentrations decreased exponentially after UV exposure, with decay constants of 0.0259 in plasma and 0.0528 in saline. The recovery of 8-MOP in photopheresates was about 68%, indicating binding to DNA as well as to plastic structures. UVA induced no 8-MOP fragmentation, but caused self-adducts under extreme conditions (10-fold UV dosage). Conclusions : Detection of 8-MOP proved to be feasible and demonstrated that the doses were in the pharmaceutically active range. An LC-MS/MS assay for analysis of 8-methoxypsoralene was developed as assay to monitor extracorporeal photopheresis. This allows quantification of 8-MOP adhering to plastic surface and of the UV light-dependent decay constant.
ISSN:1474-905X
1474-9092
DOI:10.1039/c6pp00327c