Simultaneous determination of bovine +/--lactalbumin and ^2-lactoglobulin in infant formulae by ultra-high-performance liquid chromatographymass spectrometry
A reliable ultra-high-performance liquid chromatography-mass spectrometry method for simultaneous determination of bovine @a-lactalbumin (@a-La) and @b-lactoglobulin (@b-Lg) was developed. Compared to the previous methods, the developed approach with mass spectrometer operated in selected area monitoring mode offered increased speed and enhanced lower detection limit. A linear gradient mobile phase, consisting of (A) water containing 0.1% trifluoroacetic acid (TFA) and (B) acetonitrile containing 0.1% TFA, and an Acquity UPLC BEH300 C18 column (150mmx2.1mm, 1.7@mm) were employed to obtain the best resolution of the target analytes. The accurate quantitation was achieved by employing human @a-lactalbumin as the internal standard. The established method was extensively validated by determining the linearity (R^2>=0.9991), sensitivity (limit of quantitation, 0.15-0.19@mgmL^-^1), recovery (94.0-98.7%), precision (relative standard deviation@?11.1%) and repeatability (relative standard deviation@?5.7%). It was shown to be a suitable method for simultaneous determination of the major whey proteins in biological samples. Current validated method was successfully applied to the nutrient investigation of infant formulae.
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