Method of Chromatography prof. aza Department of Pharmacy, Andalas University STIFI Perintis STIFAR Pekan Baru STIFI Bhakti Pertiwi
Hypothetical representation of chromatographic separation (a-d). Separation within the column; successive stages during elution; (e) final chromatogram corresponding to elution of sample from the column at a later time
Characteristic of a Gaussian band
Band asymmetry, (a) Definition of asymmetry factor, As; (b-e) examples of band asymmetry
Separation of two bands as a function of resolution (Rs) and relative band size (I/1, ¼, 1/16).
The basic measurement
Effects of a change in k’, N, or a on the resolution of two bands
Peak resolution R = 2d/(w 1 + w 2 )
Relationship of mean retention time (t mean ) and observed (mode) retention time (t R to EMG time constant (t)
Effect of a change in k’ (by varying solvent strength) on separation of a 5-component sample by reversed- phase LC. (a-e) 70%, 60%, 50%, 40%, and 30% aq.methanol as mobile phase,
Linear dependence of log k’ on reciprocal temperature. GC separation on Apiezon L
Linear dependence of log k’ on reciprocal temperature. LC separation on a C18 column--, Fused ring aromatic hydrocarbons; -*-, phenyl-substituted aromatic hydrocarbons; ****, 9,10- dimethyl-3,4- benzaantracene
Dependence of retention on pH in the reversed- phase LC separation of various acidic, neutral, and basic compound Separation of several bile acids
Separation of (1) salicylic acid (strong acid), (2) phenobarbital (weak acid), (3) phenacetin (neutral), (4) nicotine (weak base), (5)methylamphet amine (basa kuat).