Agilent Manuals

This experiment report introduces the effect of column length on efficiency and resolution. When appropriate, two or more SEC columns may be connected in series to achieve a desired separation.

This document investigates the effect of mobile-phase ionic strength on neutral and strongly basic antibodies. The results show that strongly basic molecules interact with silica-based column packings to a much greater extent than neutral molecules. Additionally, the concentration of phosphate buffer has an impact on peak shape and retention time.

The document introduces a high-resolution chiral separation technology for Fluoxetine (Prozac) enantiomers using the ULTRON ES-OVM column

This document introduces a method for separating immunoglobulins using size-exclusion chromatography. The method uses a high-performance ZORBAX particle that is extremely rugged, even under high back-pressures. This characteristic allows higher flow rates and shorter run times for high-throughput experimentation. The method can also reduce the injection volume of an antibody sample without loss of sensitivity, when using smaller-diameter columns.

This document introduces ZORBAX GF-250 column in the separation of Bio-Rad molecular weight markers

This document describes how to calibrate a ZORBAX GF-250 column using Bio-Rad gel filtration standards.

This document describes the complete separation of ß-amyloid peptides (ßAP) using Agilent ZORBAX 300 SB-C8 column. The inset box shows the high recovery following the HPLC separation, as determined by direct protein assay of collected peaks. The separation of these peptides was not possible at column temperatures below 80°C. High-resolution separation of human amyloid Aß peptides highlights: • Sterically-protected bonded phases (StableBond) permit use of low pH mobile phases, even at high temperature. • Use of elevated column temperature can dramatically improve separation and recoveries of difficult samples. • Note the greater retention of ßAP(1- 42) than ßAP(1-43). Application Biochemical Robert Ricker Conditions: ZORBAX 300 SB-C8, 4.6 mm ID x 250 mm, Agilent Part No. 880995-906 Mobile Phase: 80°C; 0.1% TFA in 30.5% acetonitrile / 69.5% water 2.5 µg each peptide in 20 µL 6 M urea / 5% HOAc Flow: 1 mL/min. Absorbance (210 nm) ßAP(1-38) ßAP(1-43) ßAP(1-40) ßAP(1-42) Time (min.) Rs = 2.52 0 4 8 12 16 20 Recovery (% ± S.D., n = 3) 1-38 111 ± 14 1-40 96 ± 14 1-42 75 ± 10 1-43 69 ± 10 Copyright© 2002 Agilent Technologies, Inc. All Rights Reserved. Reproduction, adaptation or translation without prior written permission is prohibited, except as allowed under the copyright laws. For more information on our products and services, visit our website at: www.agilent.com/chem Robert Ricker is an application chemist based at Agilent Technologies, Wilmington, Delaware. Agilent shall no

This is an example of using high temperature reversed-phase liquid chromatography to separate and purify amyloid peptide ab (1-42). This method takes advantage of isocratic separation (no re-equilibration time) using highly efficient 3.5 µm particle diameter, 300Å pore size, Agilent ZORBAX SB-C18 column packing material. High temperature separation enhances recovery and improves peak shape for this badly behaved peptide.

The document describes a method for high-performance liquid chromatography (HPLC) analysis using ion-pair reversed-phase separations combined with electrochemical detection to measure the levels of catecholamines and indoleamines in brain tissue. The method is rapid, selective, and efficient, allowing for analysis of samples in less than 10 minutes per sample. The use of a highly stable column-packing material (Agilent ZORBAX SB-C8, 3.5 µm) ensures unmatched column lifetime, enabling analysis of thousands of samples per column. Samples are directly injected as perchloric acid (0.1N) extracts, eliminating the need for sample workup. This method is suitable for the determination of various biogenic amines neurotransmitters and their metabolites.

The document describes the common analysis method for deoxynucleosides and presents a rapid analysis method suitable for LC-MS. The method utilizes Agilent ZORBAX SB-CN column for separation, providing good selectivity and resolution.

This document introduces the experimental method of separating the recombinant human growth hormone trypsin digestion fragments by using Agilent ZORBAX 300SB-C18, 5µm, 1 mm x 150 mm microbore column.

Size-exclusion chromatography is a powerful technique for the size-dependent separation of biomolecules. While standard, non-denaturing mobile phases are often desirable, denaturing mobile phases containing guanidine hydrochloride (or SDS), may be used to insure un-aggregated, monomers for proper size determination. As shown below, denaturation shifts molecules toward larger apparent size and smaller retention volume.

Size-exclusion chromatography is a powerful technique for the size-dependent separation of biomolecules. While standard, non-denaturing mobile phases are often desirable, denaturing mobile phases containing guanidine hydrochloride (or SDS), may be used to insure un-aggregated, monomers for proper size determination. As shown below, denaturation shifts molecules toward larger apparent size and smaller retention volume.