Development of a stable isotope dilution LC-MS assay for the quantitation of multiple polyethylene glycol (PEG) homologues to be used in permeability studies.
A new quantitation method based on a multiple stable isotope dilution assay (SIDA) was developed for polyethylene glycol (PEG) homologues from PEG mixtures with average molecular weights (MW) of 400, 1500, 3000 and 4000Da in urine. Seven [(13)C4(2)H4] and two [(13)C8(2)H8]PEG homologues were synthesized and served as labelled internal standards for SIDA. PEG oligomers were resolved by reversed phase high performance liquid chromatography (RP-HPLC) coupled to mass spectrometry (MS) in multiple ion (MI) scan modus. Very low limits of detection (LODs) in a range of 0.4-12ng/mL were achieved for the single homologues. Higher PEG homologues showed increased LODs and LOQs and less effective recovery (77-87%) than PEG with lower molecular masses (95-121%). Precision (relative standard deviation) varied between 3 and 13% and showed no dependence of the chain length. The method was successfully applied to human and mice urine samples. Beside an accurate quantitation of single PEG homologues it was possible to show an alteration in the MW distribution in urine samples compared to the dosed PEG solutions. The highest MW, with which a PEG can pass the intestinal wall (so called "cut off") for humans appeared to be higher than for mice.
Chair of Analytical Food Chemistry, Technische Universität München, Alte Akademie 10, D-85354 Freising, Germany.
Martina
M
Lichtenegger
Chair of Analytical Food Chemistry, Technische Universität München, Alte Akademie 10, D-85354 Freising, Germany; BIOANALYTIK Weihenstephan, ZIEL Research Center for Nutrition and Food Sciences, Technische Universität München, Alte Akademie 10, D-85354 Freising, Germany. Electronic address: michael.rychlik@tum.de.
Michael
M
Rychlik
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007554:Isotopes; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D010539:Permeability; D011092:Polyethylene Glycols / Q000032:analysis*; D015203:Reproducibility of Results
D007554:Isotopes; D011092:Polyethylene Glycols
Human and mice urine; Membrane permeability; PEG reference compounds; Polyethylene glycols; Quantitative NMR; Stable isotope dilution assay