Effects of Sample Deposition Medium and Drying on Spectroscopic Quantification of Lipid Biomarkers in Respiratory...

Rapid point of care assessment of pulmonary surfactant composition by measuring the lecithin/sphingomyelin (L/S) ratio could improve management of patients with neonatal respiratory distress syndrome (nRDS). Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) offers a practical route to making such measurements, but the influence of the sample solvent prior to drying on measurement repeatability is poorly understood. We compare films dried from dichloromethane (DCM) and water (AQ) solvents (DCM-dry route vs. AQ-dry route) by ATR-FTIR and show that spectra from the AQ-dry route increased the signal-to-noise ratio (SNR) of a representative (2920 cm^-1) absorption peak for the mixture from 20.13 to 128.20 and for human endotracheal aspirate (ETA) from 6.33 to 8.13. A mixed nested analysis of variance (ANOVA) showed that drying route accounted for 89.52% of mixture peak height variance and reduced percent relative standard deviation (%RSD) from 23.5% to 16.2%, corroborated by multivariate analysis for ETA. We further demonstrate that partial least squares regression (PLSR) models trained on AQ-dry mixture spectra predicted L/S (R² = 0.91; root mean square error (RMSE) = 0.31) with 95% prediction interval grey-zone interpretation around L/S = 2.2, complemented by a receiver operating characteristic area under the curve (ROC-AUC) of 0.978.