使用反相高效液相色谱法和带电气溶胶检测法进行聚乙二醇定量分析。

PEG Quantitation Using Reversed-Phase High-Performance Liquid Chromatography and Charged Aerosol Detection.

作者信息

Smith Mackensie C, Clogston Jeffrey D

机构信息

Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.

出版信息

Methods Mol Biol. 2018;1682:49-55. doi: 10.1007/978-1-4939-7352-1_5.

DOI:10.1007/978-1-4939-7352-1_5

PMID:29039092

Abstract

This chapter describes a method for the quantitation of polyethylene glycol (PEG) in PEGylated colloidal gold nanoparticles using a reversed-phase high-performance liquid chromatography (RP-HPLC) with charged aerosol detection. The method can be used to calculate the total PEG on the nanoparticle, as well as the bound and free unbound PEG fractions after a simple centrifugation step. This is a significant distinction as the bound PEG fraction affects biocompatibility, circulation time, and overall nanoparticle efficacy. PEG quantitation can be achieved through two methods, one involving the dissolution of colloidal gold nanoparticles by potassium cyanide (KCN) and the other by displacement of PEG by dithiothreitol (DTT). The methods outlined herein were applied to 30 nm colloidal gold grafted with 20 kDa PEG, but they can be easily adapted to any size colloidal gold nanoparticle and PEG chain length.

摘要

本章介绍了一种使用带电荷气溶胶检测的反相高效液相色谱（RP-HPLC）对聚乙二醇化胶体金纳米颗粒中的聚乙二醇（PEG）进行定量的方法。该方法可用于计算纳米颗粒上的总PEG，以及在简单离心步骤后结合的和游离未结合的PEG部分。这是一个重要的区别，因为结合的PEG部分会影响生物相容性、循环时间和纳米颗粒的整体功效。PEG定量可通过两种方法实现，一种是用氰化钾（KCN）溶解胶体金纳米颗粒，另一种是用二硫苏糖醇（DTT）置换PEG。本文所述方法应用于接枝20 kDa PEG的30 nm胶体金，但它们可轻松适用于任何尺寸的胶体金纳米颗粒和PEG链长度。