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1 compound A moves most rapidly and leaves the column first; compound C moves the slowest and leaves the column last. As a result, compounds A and C gradually become separated as they move through the column. Differential migration is the basis of separation in chromatography; without a difference in migration rates for two compounds, no separation is possible. 2 The basis of retention in LC. 18 Basic Concepts and Control of Separation and the flowing solvent or mobile phase. 2, for a single particle of column packing and compounds A and C.

2. 7). Often we require only a rough estimate of k', in which case k' can be determined by simple inspection of the chromatogram, without exact calculation. The important column parameter t0 can be measured in various ways. , Figure. 5 min) marks t0. If there is any doubt on the position of t0, a weaker solvent (or other nonretained compound) can be injected as sample, and its tR value will equal t0. A weaker solvent provides larger k' values and stronger sample retention than the solvent used as mobile phase, (see Chapters 6-11 for lists of solvents according to strength).

Pryde and R. A. Wall, High-Performance Liquid Chromatography, Edinburgh Univ. Press, Edinburgh, 1978 (a 212-page manual). Parris, N. , Instrumental Liquid Chromatography: A Practical Manual, Elsevier, New York, 1976 (a well-reviewed book aimed at the practical worker; Chap. ). Perry, S. , R. Amos, and P. I. Brewer, Practical Liquid Chromatography, Plenum, New York, 1972 (out-of-date and contains little that is relevant today). , and M. T. ). Rajcsanyi, P. , and E. Rajcsanyi, High Speed Liquid Chromatography, Dekker, New York, 1975 (applications oriented but does not cover ion-exchange or size-exclusion chromato graphy).

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