Amino Acids In Dried Blood Spots Research Articles

Dried blood spot reference intervals for 18 amino acids in a 1 to 6 years old pediatric cohort

ObjectivesReference intervals are indispensable for the accurate clinical interpretation of clinical laboratory tests. The reference intervals of amino acids in dried blood spots (DBS) from nonnewborn children are limited. In this study, we aim to establish the pediatric reference intervals for amino acids in DBS from healthy Chinese children aged from 1 to 6 years and to investigate the effect of sex and age. Design and MethodsIn 301 healthy subjects aged from 1 to 6 years old, eighteen DBS amino acids were determined using ultra-performance chromatography-tandem mass spectrometry method. Amino acid concentrations were examined in relation to sex and age. Reference intervals were established according to the CLSI C28-A3 guidelines. ResultsReference intervals of 18 amino acids bounded by the 2.5 and 97.5 percentiles were calculated in DBS specimen. No significant influence of age was observed for all the target amino acid concentrations in 1- to 6-year-olds. Sex differences were found in leucine and aspartic acid. ConclusionsThe RIs established in the present study added value for diagnosing and managing the amino acid-related diseases in the pediatric population.

Validation of Accuracy-based Amino Acid Reference Materials in Dried-Blood Spots by Tandem Mass Spectrometry for Newborn Screening Assays

Advances in technology and the earlier release of newborns from hospitals have pressed the demand for accurate calibration and improved interlaboratory performance for newborn screening tests. As a first step toward standardization of newborn screening aminoacidopathy tests, we have produced six-pool sets of multianalyte dried-blood-spot amino acid reference materials (AARMs) containing predetermined quantities of five amino acids. We describe here the production of the AARMs, validation of their amino acid contents, and characterization of their homogeneity and their stability in storage. To each of six portions of a pool of washed erythrocytes suspended in serum we added Phe (0-200 mg/L), Leu (0-200 mg/L), Met (0-125 mg/L), Tyr (0-125 mg/L), and Val (0-125 mg/L). Six-pool sets (1300) were prepared, dried, and packaged. We used isotope-dilution mass spectrometry to estimate the endogenous amino acid concentrations of the AARMs and validate their final amino acid concentrations. We used additional tandem mass spectrometry analyses to examine the homogeneity of amino acid distribution in each AARM, and HPLC analyses to evaluate the stability of the amino acid contents of the AARMs. The absolute mean biases across the analytic range for five amino acids were 2.8-9.4%. One-way ANOVAs of the homogeneity results predicted no statistically significant differences in amino acid concentrations within the blood spots or within the pools (P >0.05). Regression slopes (0 +/- 0.01) for amino acid concentrations vs storage times and their P values (>0.05) showed no evidence of amino acid degradation at ambient temperatures, 4 degrees C, or -20 degrees C during the intervals tested. The validation, homogeneity, and stability of these blood spots support their use as a candidate national reference material for calibration of assays that measure amino acids in dried-blood spots.