Agilent Isolation of Basic Amines in the Presence of Acidic and Neutral Impurities from Human Plasma Using Silica-Based SCX (Strong Cation Exchange), Bond Elut SCX and SampliQ Si-SCX SPEs 说明书用户手册.PDF

Isolation of Basic Amines in the Presence of Acidic and Neutral Impurities from Human Plasma Using Silica-Based SCX SPEs Technical Overview Introduction A comparison study involving two silica-based, strong cation exchange (SCX) solid phase extraction (SPE) products was conducted, focusing on the recovery of two basic amines. In addition, one acid and two neutral impurities were spiked into human plasma to evaluate the selectivity for amines only. A five-component solution was spiked into human plasma containing 2% H PO . 3 4 The sample was then loaded onto a preconditioned SPE cartridge. The SPE cartridge was washed with 2% formic acid, then with methanol/acetonitrile (1:1) to remove any organic, nonbasic components. The basic components were then eluted with 5% NH OH/(methanol/acetonitrile (1:1)). 4 HPLC analysis of the SPE extracts used an Agilent ZORBAX Rapid Resolution Eclipse Plus C18 column with a gradient of 20 to 90% methanol/10 mM ammonium formate (pH 3.8). Both Agilent Bond Elut SCX and SampliQ Si-SCX showed comparable and consistent recoveries for the basic compounds used in this study. The extractions presented in this technical note suggest that the use of Bond Elut SCX (silica based SCX) gives a clean extract and high recovery of basic compounds, such as amines. Study Purpose and Methodology The purpose of the study was to evaluate performance differences between Agilent Bond Elut SCX and Agilent SampliQ Si-SCX, which are silica-based strong cation exchange SPE products, with respect to their retention of basic amines. The recov- eries of two basic components and the chromatographic profile of the eluents are evaluated to determine performance differences. Procainamide and chlorpromazine were chosen as representative bases. One acid and two neutral compounds were added to the base spiking solution as representa- tive acid and neutr