Development of Flow Injection Analysis Method for the Second-Tier Estimation of Succinylacetone in Dried Blood Spot of Newborn Screening

AbstractTyrosinemia type 1 (TYR1) is a devastating aminoacidopathy, leading to mortality without medical intervention. Although, detection and quantification of tyrosine in dried blood spot (DBS) is possible, but being a non-specific marker for TYR1 and its frequent association with transient neonatal tyrosinemia limits its applicability. Despite, Succinylacetone (SUAC) being a pathognomonic marker for TYR1, but not often detectable by routine newborn screening (NBS). We envisaged to determine SUAC in DBS by an in-house flow injection analysis method on a liquid chromatography/tandem mass spectrometry (LC –MS/MS). Succinylacetone was eluted from the residual 3.2 mm DBS of primary NBS by an extraction solution containing acetonitrile–water–formic acid mixture containing stable-isotope labelled internal standard (IS) for SUAC and hydrazine. Detection and quantification was performed by the mass spectrometer using multiple reaction monitoring mode at m/z 155.1 → 109.1 for SUAC and m/z 160.1 → 114.1 for the SUAC IS. The assay was linear over a calibration range of 0.122–117.434 µmol/L. The Intra-day and Inter-day precision and accuracy for the assay was determined at two diff erent levels of SUAC (2.542 µmol/L and 14.641 µmol/L), which showed a coefficient of variation of (6.91% and 12.65%) and (8.57% and 12.27%) respectively. The accuracy also ranged between 101.2 and 103.87%.This method provided the necessary sensitivity, precision, accuracy, recovery...
Source: Indian Journal of Clinical Biochemistry - Category: Biochemistry Source Type: research