Mutalik S, Hewavitharana A K, Shaw P N, Anissimov Y G, Roberts M S, Parekh H S
School of Pharmacy, University of Queensland, Brisbane, Qld 4072, Australia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Nov 1;877(29):3556-62. doi: 10.1016/j.jchromb.2009.08.039. Epub 2009 Aug 29.
The aim of the present work was to develop and validate a simple RP-HPLC method with UV detection to quantify peptide dendrimers in skin permeation experiments. Six dendrimers of varying positive charges (4(+), 8(+) and 16(+)) containing either histidine or arginine as terminal aminoacids were prepared by solid phase peptide synthesis. Mobile phase containing 0.02% (v/v) heptafluorobutyric acid in 90% acetonitrile-water was capable of separating all dendrimers from interfering peaks of receptor fluid. For the calibration of each dendrimer, a different dendrimer from the same class was selected as the internal standard. The results of preliminary human skin permeation studies showed that the developed analytical method can be successfully used for the quantification of cationic poly(aminoacid)-based dendrimers in skin permeation experiments.
本研究的目的是开发并验证一种简单的带有紫外检测的反相高效液相色谱法，用于在皮肤渗透实验中对肽树枝状大分子进行定量分析。通过固相肽合成法制备了六种带有不同正电荷（4(+)、8(+)和16(+)）的树枝状大分子，其末端氨基酸为组氨酸或精氨酸。流动相为在90%乙腈-水中含有0.02%（v/v）七氟丁酸，能够将所有树枝状大分子与受体液的干扰峰分离。对于每种树枝状大分子的校准，从同一类别中选择一种不同的树枝状大分子作为内标。初步人体皮肤渗透研究结果表明，所开发的分析方法可成功用于皮肤渗透实验中基于阳离子聚氨基酸的树枝状大分子的定量分析。
