Analytical method for the determination of polyethylenglycole 400 as marker in porcine plasma

Polyethylenglycole (PEG) is a widespread linear polymer which can be utilized as a solute digestive and intestinal permeability marker in nutritional physiology studies depending on chain length/molecular mass. PEG 400 is proposed to be an ideal permeability marker. Due to its molecular mass (238-590 g/mol) and characteristics, PEG 400 is suggested to be used as a surrogate for studying the paracellular permeability of small hydrophilic molecules. For this purpose, a liquid chromatographic-tandem mass spectrometric method has been developed for the determination of the major oligomers of PEG 400 in porcine plasma. The analysis included a simple and rapid clean-up step where proteins were precipitated. The most intense ions corresponding to seven PEG 400 oligomers were separated within 7 min. Validation of the optimized method was performed in the range of 500-18,000 ng/mL. Mean recoveries between 93 and 105% were achieved using spiked plasma samples in three different concentration levels. The limit of quantification ranged between 11 and 244 ng/mL. The applicability of the method was demonstrated by the analysis of porcine plasma samples obtained from an animal experiment with barrows. The kinetic course of administrated PEG 400 was shown based on the dataset of two barrows selected from the control group, and it was figured out that relative proportion of each PEG oligomer in portal plasma decreased with increasing molecular mass.