Download HPLC: A Practical User's Guide by Marvin C. McMaster PDF

By Marvin C. McMaster

This moment version of the vintage instruction manual info how you can arrange an HPLC process that capitalizes at the most recent recommendations. It covers new strategies in high-temperature, micro-flow, and ultra-fast chromatography, the linking of an HPLC to a mass spectrometer, and extra. whole with a CD-ROM and appendices, this advisor has every thing chromatographers want to know to with a bit of luck separate, determine, purify, and quantify compounds.

Note: CD-ROM/DVD and different supplementary fabrics usually are not incorporated as a part of booklet dossier.

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Additional resources for HPLC: A Practical User's Guide

Example text

If no peaks appear after 5 min at final conditions, the water is good. The chromatogram (Fig. 4) gives you an idea of the expected baseline appearance. Peaks that appear during the first acetonitrile washout are ignored as impurities already on the column. Watch the baseline on switching to water. At 254 nm, the baseline should gradually elevate. If instead it drops, you may have impurities in your acetonitrile. If the baseline makes a very sharp step up before leveling off, you may have a large amount of polar impurities in the water.

He blamed the problem on the detector. I said it looked like bad water. He exploded, told me that his water was triple distilled and good enough for enzyme reactions. It was good enough for HPLC, he said. Over the following 6 mo we replaced 32 RUNNING YOUR CHROMATOGRAPH every component in that system as each in turn was blamed for the chromatography problem. Eventually, the customer borrowed HPLC-grade water from another institution, washed his column with acetonitrile, then with water. The problem disappeared and never came back—until he went back to his own water.

Some also can be calibrated with compound names related to peak retentions to provide annotated outputs. Integrating systems are designed to make the chromatographer’s life easier, but they can complicate it if not properly used. They usually have an auto/zero function, which, when selected, looks at the baseline before injection and sets various integration parameters. This is designed to prevent integration of very small or extraneous peaks or of baseline noise. On most integrators, autozero must be requested by the operator and should be used every time a detector attenuation change is made.

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