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Bioptic AV-1 and AV-2 Columns for Direct Injection of Plasma |
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Glycoprotein bonded silica allows direct injection of plasma samples without
protein removal |
Plasma proteins elute in void volume |
Small molecules elute after protein void peak with reverse phase type
separation |
Bioptic AV-1 allows chiral separation of many drug compounds |
Large sample capacity |
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Analysis of drugs in plasma has
traditionally required sample preparation to remove plasma proteins before
HPLC analysis to avoid precipitation of plasma proteins and subsequent
column clogging.
Bioptic AV columns allow the inner hydrophobic surface of the silica to be
shielded by a permanently bonded hydrophilic avidin layer. Large
molecules like plasma proteins are excluded from interacting with the
hydrophobic inner surface by the avidin layer, and elute by size-exclusion
mechanisms in the void volume. Smaller compounds, such as drugs and
drug metabolites, pass through the avidin layer and interact with the
hydrophobic inner layer by a reverse-phase mechanism.
Bioptic AV-1 contains
non-denatured avidin and a chiral center, allowing separation of chiral
drugs and their metabolites. Bioptic AV-2 contains a denatured form of
avidin and lacks the chiral separating ability of AV-1, but offers different
selectivity.
Bioptic columns are made with a 5 micron spherical silica, and the
columns withstand pressures up to about 2250 PSI. AV-1 has a limit of
about 20% on the organic content of the eluent, which there is no such
limitation on AV-2. Recommended pH range is 2 to 7.5, while pH ranges where
hydrophobic interactions occur are between 2-3.5 and between 6 and 7.5.
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