Biopolymers & Proteomics Laboratory

The David H.Koch Institute for Integrative Cancer Research at MIT

Bldg 76 Room 181

Telephone: 617-253-7038

HPLC

Introduction

Since the MIT Center for Cancer Research merged with MIT Bioengineers doing cancer research forming the new KIICR, we saw a need to assist if possible in the purification of compounds created by this new entity, most of which are related to drug delivery or measurement.

Preparative HPLC

High Performance (a.k.a. high pressure) Liquid Chromatography (HPLC) offers high resolution separation of compounds from a complex mixture. Amounts of approximately 1 mg and upward are analyzed on the Gilson model 271 system with UV-visible absorption wavelength detection. After column fractionation, separated components are saved in liquid form in fraction collector tubes for further use. We can MALDI-TOF or ESI mass analyze fractions and pool, speedvac or lyophilize fractions that have the correct molecular weight and are homogeneous by analytical HPLC. In addition, side fractions can be saved and rerun later to obtain more pure material.

Typical analytes are peptides, proteins, small molecules, conjugates, RNA/DNA, drugs, polymers and dye-labeled compounds. Bring a reference or at least the molecular structure to help us chose LC column and mobile phase conditions.

Analytical HPLC

We have three Agilent model 1100 systems with autosamplers and UV-visible detectors for analytical and small-scale preparative (under 1 mg) purposes available for the analysis of your material. Up to eight absorption wavelengths can be monitored at once plus a diode array scan from 200 to 700 nm each second throughout the run. We generally use reverse phase columns like C1, C4 and C18 with acetonitrile, methanol or isopropanol gradients at pH of about 3 or 7.

If you presently have access to a LC system, we can advise you regarding solvent systems, etc. If you have perhaps hundreds of samples to analyze or need to trail blaze methods we can train you on these instruments and you can “rent” an Agilent model 1100 located in the Nanomaterials Technology Core.

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