Mid-infrared predictions of lactoferrin content in bovine milk

Lactoferrin (LF) is a glycoprotein present in milk and active in the immune system of cows and humans. Therefore, an inexpensive and rapid analysis to quantify this protein is desirable. A previous study reported the potential to quantify LF from the mid-infrared (MIR) spectrometry from 69 milk samples. Through the European RobustMilk project (www.robustmilk.eu), 3,606 milk samples were collected in Belgium, Ireland, and Scotland from individual cows and analyzed using a MIR MilkoScanFT6000 spectrometer. Milk LF content was quantified using ELISA in duplicate. Average ELISA data with a CV lower than 5% were used. After the detection of spectral and ELISA outliers, the calibration set contained 2,499 samples. An equation to predict LF content from MIR was developed using partial least squared regression. A first derivative pre-treatment of spectra was used to correct the baseline drift. To improve the repeatability of the spectral data, a file which contained the spectra of samples analyzed on 5 spectrometers was used during the calibration. The lactoferrin mean was 159.28 mg/l of milk with a SD of 97.21 mg/l of milk. The calibration (C) coefficient of determination (R2) was equal to 0.73 with a standard error (SE) of calibration of 50.54 mg/l of milk. A cross-validation (CV) was used to assess the robustness of the equation. R2 CV was 0.72 with a SE-CV of 51.16 mg/l of milk. An external validation (V) was conducted on 150 milk samples collected in Belgium. The SE of prediction (SEP) was 59.17 mg/L of milk. The similarity between R2 C and R2CV as well as between SE-C and SE-CV and between SE-CV and SEP confirms the equations developed are robust. The correlation between predicted and measured LF values was 0.71. This lower value compared with the one obtained from the calibration set (0.85) could be explained by the low ELISA reproducibility (16.24% ± 25.51%). If the developed equation is used to clean the validation data set, a total of 16 samples can be deleted. The validation coefficient for these 134 samples increased to 0.82. From these results, the developed equation could be used for screening the dairy cow population for breeding purposes.Robustmil