Biomarker Profiling for Pyridoxine Dependent Epilepsy in Dried Blood Spots by HILIC-ESI-MS

Mathew, Elizabeth Mary and Moorkoth, Sudheer and Lewis, Leslie Edward Simon and Rao, Pragna (2018) Biomarker Profiling for Pyridoxine Dependent Epilepsy in Dried Blood Spots by HILIC-ESI-MS. International Journal of Analytical Chemistry, 2018. pp. 1-8. ISSN 1687-8760

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Abstract

Pyridoxine dependent epilepsy is a condition where the affected infant or child has prolonged seizures (status epilepticus), which are nonresponsive to anticonvulsant therapy but can be treated with pharmacological doses of pyridoxine. If identified earlier and treated prophylactically with pyridoxine, severe brain damage due to seizures can be prevented. Alpha-amino adipic semialdehyde (AASA), piperidine-6-carboxylic acid (P6C), and pipecolic acid (PA) are known biomarkers of pyridoxine dependent epilepsy.We report the development and validation of a hydrophilic interaction liquid chromatography (HILIC) hyphenated with mass spectroscopy for the quantification of the above analytes fromdried blood spot samples.The samples were extracted usingmethanol and analysed on a iHILIC fusion plus column with formic acid buffer (pH 2.5): acetonitrile (20:80) at a flow rate of 0.5mL/min within 3 minutes. The method demonstrated a LOD of 10 ng/mL, LOQ of 50 ng/mL, linearity of r2 ≥ 0.990, and recovery of 92-101.98% for all analytes. The intra- and interday precision CVs were < 8% and 6%, respectively. Extensive stability studies demonstrated that the analyteswere stable in stock solution and inmatrixwhen stored at -80∘C.We performedmethod comparison studies of the developed method with the literature reported method using normal samples and matrix matched spiked samples at pathological concentrations tomimic clinical validity.The Bland-Altman analysis for comparison of the analytical suitability of the method for the biomarkers in healthy and spiked samples with the literature reported method revealed a bias which suggested that the method was comparable.The newly developed method involves no derivatisation and has a simple sample preparation and a low run time enabling it to be easily automated with a high sample throughput in a cost-effective manner.

Item Type: Article
Subjects: Medicine > KMC Manipal > Biochemistry
Medicine > KMC Manipal > Paediatrics
Pharmacy > MCOPS Manipal > Pharmaceutical Quality Assurance
Depositing User: KMC Library
Date Deposited: 22 Nov 2018 09:17
Last Modified: 22 Nov 2018 09:17
URI: http://eprints.manipal.edu/id/eprint/152326

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