In this article, we describe the synthesis of a new class of oligoethylene-glycol based water-soluble biotin derivatives for labeling of peptides with limited solubility in aqueous solution. First 4,7,10-trioxa-1,13-tridecanediamine was mono-acetylated by succinic anhydride (Ttds) followed by the introduction of N-Fmoc-protecting group using Fmoc-N-hydroxysuccinimide ester. The resulting compound (Fmoc-Ttds) was used for the preparation of 4,7,10-trioxa-1,13-tridecanediamine di- and trimers on solid phase using Wang resin by carbodiimide coupling method. After attachment of Fmoc-Ttds to the solid support, the Fmoc-blocking group was removed and the Ttds-modified resin was repeatedly acylated by Fmoc-Ttds or by biotin using PyBOP/HOBt active ester reaction. Finally the product [Fmoc-(Ttds)(n) or biotinyl-(Ttds)(n) (where n = 1, 2 or 3)] was removed from the resin by trifluoroacetic acid in the presence of water. After appropriate HPLC purification and characterization (MS) biotinyl-(Ttds)(n) (where n = 1, 2 or 3) were introduced to the N-terminal of poorly soluble oligopeptides by solid phase peptide synthesis. We found that this new class of biotinylating reagent could be prepared easily and in good yield. Comparative solubility measurements suggest that the incorporation of these moieties-depending on the number of Ttds unit-could enhance water solubility.