BackgroundInfrared (IR) analysis is an emerging technology that may be a useful tool for milk banks to manage the nutrient variability in donor human milk.ObjectiveTo evaluate the accuracy, reliability, and comparability of commercial infrared analyzers for measuring human milk macronutrients in a milk bank setting.MethodsThree nonprofit milk banks received blinded test kits of human milk that had been assessed using reference methods. Four infrared instruments were used to measure macronutrients as follows: 1 filtered mid-IR, 2 Fourier-transformed full-spectra mid-IR, and 1 near-IR. Twenty-five unique samples were read concurrently for the accuracy arm. An identical sample was read daily for 1 mo for the reliability arm.ResultsValues for R2 describing relationships with reference methods for total fat, crude protein, and lactose, were as follows: filtered mid-IR, 0.98, 0.94, and 0.48; Fourier-transformed full-spectra mid-IR, 0.97, 0.93, and 0.36 for instrument 1 and 0.98, 0.98, and 0.31 for instrument 2; and near-IR 0.93, 0.93, and 0.12. There was no significant difference between instruments for crude protein and total fat measurements. There were significant differences in carbohydrate measurements between instruments. For 1 mo of daily measurements in the reliability arm, CVs for filtered mid-IR were ≤4.6%, for Fourier-transformed full spectra mid-IR were ≤1.7%, and for near-IR were ≤5.1%.ConclusionsInfrared analysis is an accurate and reliable method for measuring crude protein and total fat in a milk bank setting. Carbohydrate measurements are less accurate and are significantly different between instruments, which will likely lead to differences in derived calorie values.