Lorek Miłosz, Stradomska Teresa Joanna, Siejka Anna, Fuchs Janusz, Januś Dominika, Gawlik-Starzyk Aneta
Department of Neonatal and Pediatric Intensive Care, John Paul II Center for Child and Family Health, Sosnowiec, Poland.
Department of Pediatrics and Pediatric Endocrinology, Medical University of Silesia, Katowice, Poland.
Endokrynol Pol. 2025;76(3):331-338. doi: 10.5603/ep.106090.
Steroid metabolomics in neonatal populations is challenged by considerable physiological heterogeneity and technical variability, which complicate the interpretation and comparability of metabolite profiles. Effective normalization strategies are essential to ensure accurate data analysis in this context.
We analyzed 24-hour urinary steroid profiles in a cohort of 50 neonates (including very preterm, late preterm, and full-term infants) using gas chromatography-mass spectrometry. Two normalization techniques were compared: probabilistic quotient normalization (PQN) and peer group normalization (PGN). Normalization performance was assessed via distribution metrics, correlation with anthropometric variables, and principal component analysis (PCA).
PGN achieved superior distributional normalization, with 27 of 30 metabolites conforming to normality assumptions, compared to 21 using PQN. PGN also eliminated all significant correlations between steroid levels and anthropometric parameters, indicating effective reduction of physiological confounding. In contrast, PQN partially mitigated such associations but was less robust in handling high-abundance metabolites. PCA confirmed improved sample dispersion and group separation after normalization, with method-dependent differences in Scores Plot.
Peer group normalization is a sophisticated approach to reducing physiological variability in neonatal steroid profiling. These observations lend further credence to PGN as a promising strategy for standardizing steroid metabolomics in the field of neonatology. Nevertheless, further validation is necessary to substantiate these findings.
新生儿群体中的类固醇代谢组学面临着显著的生理异质性和技术变异性挑战，这使得代谢物谱的解释和可比性变得复杂。在这种情况下，有效的标准化策略对于确保准确的数据分析至关重要。
我们使用气相色谱 - 质谱法分析了50名新生儿（包括极早产儿、晚期早产儿和足月儿）队列中的24小时尿类固醇谱。比较了两种标准化技术：概率商数标准化（PQN）和同组标准化（PGN）。通过分布指标、与人体测量变量的相关性以及主成分分析（PCA）评估标准化性能。
与使用PQN时的21种相比，PGN实现了更优的分布标准化，30种代谢物中有27种符合正态性假设。PGN还消除了类固醇水平与人体测量参数之间的所有显著相关性，表明有效减少了生理混杂因素。相比之下，PQN部分缓解了此类关联，但在处理高丰度代谢物方面不够稳健。PCA证实标准化后样本离散度和组间分离得到改善，得分图存在方法依赖性差异。
同组标准化是一种减少新生儿类固醇谱生理变异性的复杂方法。这些观察结果进一步证明PGN是新生儿学领域标准化类固醇代谢组学的一种有前景的策略。然而，需要进一步验证来证实这些发现。
