Characterization of the precolumn bio trap 500 C18 for direct injection of plasma samples in a column-switching system

Abstract

SummaryA new restricted access media (RAM) type of precolumn, BioTrap 500 C18, for direct injection of plasma samples in column-switching systems was evaluated with respect to the elution of plasma proteins in different mobile phases, the loading capacity of plasma samples, the chromatographic behavior during plasma injections and protein contamination of the packing and sealings. More than 95 % of plasma proteins could be excluded from the precolumn within three minutes for all selected mobile phases. Quantitative analyte recoveries could be obtained by injecting plasma samples ranging from 5 to 500 ~L with the analyte mass > 150 ng onto a BioTrap 500 C18 column (20 • 4 mm I.D.). One precolumn tolerated about 15 mL of plasma injection without noticeable change in retention and pressure. Clogging of the precolumn was encountered (__ 45 mL of plasma) due mainly to the adsorption of proteins on the packing. The performance of the analytical column (Kromasil C18) was also examined. The column efficiency decreased by 60 % after processing 45 mL plasma in total.

Methylcellulose-immobilized Reversed-phase Precolumn for Direct Analysis of Drugs in Plasma by HPLC

“…As a protein-immobilized stationary phase capable of plasma direct injection, octadecylsilane (ODS) treated with human plasma was developed and plasma direct injection capability was shown by Yoshida et al 12,13 α1-Acidglycoprotein (α1-AGP)-, ovomucoid-and avidin-coated columns had been developed, [14][15][16] and are capable of plasma direct injection. [17][18][19][20] Furthermore, direct analysis of drugs in serum using zwitterionic bile acid derivative-coated ODS was reported by Umemura et al 21 Several polymer-modified reversed-phase column materials such as the shielded hydrophobic phase (SHP) and the semipermeable surface (SPS) had also been reported. 8,[22][23][24] SHP consists of a hydrophilic, polymeric, water-solvated phase, covalently bonded to a silica bed.…”

Column-switching HPLC–MS/MS analysis of ropivacaine in serum, ultrafiltrate and drainage blood for validating the safety of blood reinfusion

“…Using the BioTrap as precolumn for the extraction of polar drugs from plasma or serum has been thoroughly investigated [24,25]. The BioTrap 500 MS column particles are manufactured with a coating of ␣1-acid glycoprotein (AGP) on the external surface and consequently compatible with various types of protein-containing biosamples under generic conditions for either basic or acidic compounds.…”

Solid-phase extraction in clinical biochemistry

“…Silica C 18 restricted-access matrix sorbents e¡ectively exclude plasma proteins. 33,34 A 2064 mm i.d. Bio Trap C 18 column (Chromtech) could tolerate up to 15 mL of plasma without changing retention pressure, with clogging only after 45 mL of plasma.…”