A comprehensive approach for detection of biotin deficiency from dried blood spot samples using liquid chromatography-mass spectrometry

<b>Aim:</b> The aim of the present study is to develop a liquid chromatography-mass spectrometry method to measure two important biomarkers of biotin deficiency from dried blood spot samples for effective management of the disorder. <b>Materials &amp; methods:</b> The method was developed on a liquid chromatography-mass spectrometry system using pentafluorophenyl column employing a mobile phase composition of methanol and water in the isocratic mode. A full validation of the method was performed as per relevant guidelines. <b>Results &amp; conclusion:</b> Correlation between the results of dried blood spot and plasma method was evaluated to determine the interconvertibility of the method. The developed method was successfully applied for establishing the reference ranges for these biomarkers in the population of Udupi, a coastal district of South India. Biotin deficiency can lead to many complications such as impaired growth, compromised immune function, depression, myalgia and may even lead to death. The disorder can be managed by supplementation of biotin. Early detection is crucial in managing biotin deficiency. In this paper we describe a comprehensive method for the early detection of biotin deficiency. The method employs the use of minimally invasive blood sampling such as dried blood spot that is suitable for vulnerable neonatal population. The LC-MS method targets the simultaneous determination of 3-hydroxyisovaleric acid (3-HIV) and 3-hydroxyisovaleryl carnitine (3-HIC) in dried blood spots (DBS), providing a comprehensive analysis for diagnosing and preventing biotin deficiency in newborns and infants. Simple, selective, rapid, specific and highly sensitive. The method was validated for accuracy, precision, recovery, matrix effect, hematocrit effect, stability and blood volume/punch position effects. High linearity (R2 &gt;0.99), good sensitivity (LLOQ 0.088 μM and 0.040 μM for 3-HIV and 3-HIC respectively). Recovery of QC samples was within acceptable limits for both analytes. Strong correlation and minimal bias were found between DBS and plasma measurements. Applied successfully for establishing reference ranges in the population of Udupi, South India. 3-HIV (0.06–18.59 μM) and 3-HIC (0.01–0.49 μM) in Udupi population. The newly developed method is a promising tool for confirmatory diagnosis of biotin deficiency in a clinical setting.