HPLC Method for Analysis of Polyethylenimine (PEI) on PEI Column

https://sielc.com/hplc-determination-of-pei
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Chromatogram

Description

Large multi-charged molecules such as Polyethyleneimine (PEI) can be very difficult to separate and retain on standard columns. PEI is a complex mixture of the molecules of different sizes and geometries with various biological and pharmaceutical applications, such as in tissue cultures, as a transfection reagent, and in carbon dioxide capture. PEI also lacks a UV chromophore, making it difficult to analyze with a standard UV detector.

SIELC’s new PEI column was designed specifically for the analysis of PEI by ion-exclusion and size-exclusion mechanisms. This method uses a simple isocratic mobile phase of Acetonitrile (MeCN) and water with an ammonium formate (AmFm) buffer. This method can be analyzed with high resolution and peak symmetry via any evaporative detector (including ELSD, CAD, and ESI-MS), removing the need for the use of a Copper(II) complex.

Method Parameters

Mobile PhaseMeCN/H2O – 40/60%
BufferAmmonium Formate pH 3.0 – 20 mM
Flow Rate0.5 ml/min
DetectionELSD, the nebulizer and evaporator temperatures 40°C, with a gas flowrate of 1.6 Standard Liters per Minute (SLM)
Peak Retention Time3.81 min
Class of CompoundsDrug
Analyzing CompoundsPolyethylenimine PEI

HPLC Column Used

PEI, 4.6 x 250 mm, 5 µm, 100 A, dual ended