Development and validation of a simple LC-MS/MS method for the quantification of MTHF in human cerebrospinal fluid using deuterated MTHF as internal standard

Dewulf, Joseph;Vanderstocken, Christophe;Nassogne, Marie-Cécile;Marie, Sandrine;Vincent, Marie-Françoise
(2013) International Congress of Inborn Errors of Metabolism (ICIEM) — Location: Barcelona, Spain (3.September.2013)

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Abstract
Objective: Development and validation of a LC-MS/MS method for the quantification of MTHF in cerebrospinal fluid (CSF) using deuterated MTHF as internal standard (IS) for the diagnosis of cerebral folate deficiency. Materials and methods: No pre-treatment was required and artificial CSF was chosen for the validation. The CSF was first diluted ½ with ascorbic acid IS solution to reduce ion suppression effect. The analyte and IS were eluted (same retention time) on LiCrospher RP-18 5μ 150mm x 4,6mm (Grace) using an isocratic elution method. Detection and quantification were performed by electrospray ionization (ESI) in the positive ionization mode by multiple reaction monitoring (MRM). Results: The assay was linear over the concentration range of 10-300nM (R2 >0,999) with the lower limit of quantification (LLOQ) of 10nM. The intra- and inter-day precision in terms of relative standard deviation (RSD %) was within 7, 2% and accuracy in terms of relative error (RE %) was within 13, 75%. Conclusion: This method fulfills all the regulatory requirements for specificity, linearity, LLOQ, precision and accuracy for the determination of MTHF in human CSF.
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Dewulf, J., Vanderstocken, C., Nassogne, M.-C., Marie, S., & Vincent, M.-F. (2013). Development and validation of a simple LC-MS/MS method for the quantification of MTHF in human cerebrospinal fluid using deuterated MTHF as internal standard. International Congress of Inborn Errors of Metabolism (ICIEM), Barcelona, Spain. https://hdl.handle.net/2078.5/170563