BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
BioHPLC Column Selection Guide Cover - Agilent Technologies
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Preparative scale purification of<br />
Leuprolide by concentration overload<br />
<strong>Column</strong>: PLRP-S 100Å, 10 µm<br />
PL1412-4100<br />
Bulk Media: Load & Lock 4001 <strong>Column</strong><br />
PCG93LL500X25<br />
Mobile Phase: Isocratic separation<br />
using 0.1% TFA<br />
in 28% ACN:72% water<br />
Flow Rate: Linear velocity 360 cm/hr<br />
Biomolecule Separations<br />
Table 1. <strong>Agilent</strong> columns for small- to large-scale peptide purifications.<br />
<strong>Agilent</strong> <strong>Column</strong><br />
ZORBAX Prep HT 300StableBond<br />
VariTide RPC<br />
PLRP-S<br />
Amount of Peptide Required<br />
mg g kg<br />
After solid phase synthesis (SPS) using a polystyrene resin such as one of the <strong>Agilent</strong> StratoSpheres<br />
products, the peptide is cleaved from the support and the resultant mixture is separated to obtain<br />
the target peptide. A high efficiency column is needed for the purification as the candidate peptide<br />
must be resolved from peptides that are very similar in structure. Check www.agilent.com for<br />
further information.<br />
0 Time (min)<br />
40<br />
Crude leuprolide separation of 30 mg on-column load.<br />
Fraction analysis – the concentration overload purification.<br />
% Purity (Pre-impurity 1)<br />
% Purity (Leuprolide)<br />
% Purity (Post-impurity)<br />
<strong>BioHPLC</strong> COLUMNS<br />
11
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