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Large failure rate of PEGylated pharmaceuticals in pre-clinical and clinical studies could be majorly attributed to their particular ambiguous pharmacokinetic habits. Therefore, the in vivo fate of the PEGylated pharmaceuticals for the various channels of administration has to be thoroughly investigated An accurate in vivo pharmacological study thereof highly depends on the precise detection of polyethylene glycols as well as their fragments in biological matrixes. The aim of this review