Metabolomics of a neonatal cohort from the Alliance for Maternal and Newborn Health Improvement biorepository: Effect of preanalytical variables on reference intervals

Abstract

Background The study was conducted to determine reference interval (RI) and evaluate the effect of preanalytical variables on Dried blood spot (DBS)-amino acids, acylcarnitines and succinylacetone of neonates. Methodology DBS samples were collected within 48–72 hours of life. Samples were analyzed for biochemical markers on tandem mass spectrometer at the University of Iowa. Comparison of RI across various categorical variables were performed. Results A total of 610 reference samples were selected based on exclusion criteria; 53.2% being females. Mean gestational age (GA) of mothers at the time of delivery was 38.7±1.6 weeks; 24.5% neonates were of low birth weight and 14.3% were preterm. Out of the total 610 neonates, 23.1% were small for GA. Reference intervals were generated for eleven amino acids, thirty-two acylcarnitines and succinylacetone concentrations. Markers were evaluated with respect to the influence of gender, GA, weight and time of sampling and statistically significant minimal differences were observed for some biomarkers. Conclusion RI for amino acids, succinylacetone and acylcarnitine on DBS has been established for healthy neonates, which could be of use in the clinical practice. Clinically significant effect of GA, weight, gender and time of sampling on these markers were not identified.

Targeted and untargeted metabolomics and lipidomics in dried blood microsampling: Recent applications and perspectives

“…The majority of those papers adopted DBS collection methods. [37][38][39][40][41][42] Notably, Gao et al 43 deviated from this trend by validating a method using VAMS. In the case of Carling et al, 44 .…”

“…These challenges include issues related to the volume of blood applied, the HCT effect and the quality of the sample, which had an effect on monitoring the disease. 44 Concerning sample collection in DBS assays, commonly, a subpunch of 3.2 mm in diameter was performed to collect the blood applied, 37,39,40,42 except for Zhan et al, 38 who obtained two 5 mm sub-punches. In Carling et al 44 tests, the sub-punch in conventional PerkinElmer-226 filter paper was 8 mm, while in volumetric DBSs (HemaXis™ DB10), it was 7 mm, and in Capitainer ® , it was 6 mm.…”

“…They serve as robust references for metabolic disease screening using LC-QqQ-MS detector, regardless of gender, weight or gestational age. 39 A recent study provides valuable insights into the influence of maternal lifestyle factors, drug intake and supplements during pregnancy on the metabolic profile of newborns. 42 The study included 109 mother-infant pairs and utilised NBS test to assess the impact of various factors such as smoking, alcohol consumption and physical activity.…”

“…These findings suggest that the reference intervals for these metabolites can be reliably used for interpreting DBS metabolomics data in neonates. They serve as robust references for metabolic disease screening using LC–QqQ–MS detector, regardless of gender, weight or gestational age 39 …”

Current Status of Newborn Bloodspot Screening Worldwide 2024: A Comprehensive Review of Recent Activities (2020–2023)

“…Recent project reports seeking to determine baseline information on screenable conditions found cases of HCY, MPS, and GAMT through biochemical screening of intellectually disabled patients [620]. Reports of reference range studies have included a multicenter study of TSH [621], a study of BIO [622], and a DBS study of various amino acids, succinylacetone and acylcarnitine [623]. Looking to the future, a study of an international partnership with an overseas facility performing NGS testing for PID patients suggests that resource-limited regions may require immunophenotyping and NGS in rather than TREC analysis, a screening strategy that can be met with through global partnerships [624].…”