A new analytical method has been developed for the quantitative determination of ethylene glycol-containing nonionic surfactants, such as polyethylene glycol 8000, polysorbate 80, and Pluronic F-68. These surfactants are commonly used in pharmaceutical protein preparations, thus, testing in the presence of protein is required. This method is based on the capillary gas chromatographic analysis of ethylene glycol diacetate formed by hydrolysis and acetylation of surfactants that contain ethylene glycol. Protein samples containing free surfactants were hydrolyzed and acetylated with acetic anhydride in the presence of p-toluene sulfonic acid. Acetylated ethylene glycol was extracted with dichloromethane and analyzed by gas chromatography using a flame ionization detector. The amount of nonionic surfactant in the sample was determined by comparing the released ethylene glycol diacetate signal to that measured from calibration standards. The limits of quantitation of the method were 5.0 μg/mL for polyethylene glycol 8000 and Pluronic F-68, and 50 μg/mL for polysorbate 80. This method can be applied to determine the polyethylene glycol content in PEGylated proteins or the final concentration of polysorbate 80 in a protein drug in a quality control environment.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5024075 | PMC |
| http://dx.doi.org/10.1002/jssc.201400766 | DOI Listing |
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