High Pressure Liquid Chromatography. Biochemical and by Phyllis R. Brown

By Phyllis R. Brown

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Le Rosen and Rivet (1948) and Lie Tien (1953) investigated temperature effects on solute retention in liquid chromatography. Martire and Locke (1967) studied this effect in liquid-liquid systems and Dybeznski (1967) and Horvath et al. (1967) examined temperature dependence in ion exchange chromatography. Hesse and Englehardt (1966) investigated the possibility of temperature programming and Obrink et al. (1967) and Leach and O'Shea (1965) found that temperature can affect column performance in gel permeation chromatography.

A good review of the trends in automation for Chromatographie systems was written by Anderson (1972). The use of integrators with high pressure liquid chromatographs is discussed more fully in Chapter 5. CHAPTER 3 EXPERIMENTAL METHODS High pressure liquid chromatography is becoming increasingly popular as an analytical tool because of its versatility. As in gas chromatography, the stationary phase, the flow rate, and the temperature can be varied; liquid chromatography, however, has the added advantage of being able to change many of the variables of the mobile phase.

Standardization of Conditions In order to be able to check the reproducibility of the results and the stability of instrument and column conditions, a standard solution should be analyzed routinely. The standard should be typical of the type of compounds being separated. For example, in the analysis of nucleotides, Brown (1970) analyzed a standard solution of adenine and guanine nucleotides at TV^ GDP 0 5 10 15 20 25 30 35 40 45 ATP, 50 55 60 65 70 75 TIME (MIN) Fig. 3 - 1 . Nucleotide standards.

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