Method of Chromatography

prof. azaDepartment 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/(w1 + w2)

Relationship of mean retention time (tmean) and observed (mode) retention time (tR 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)methylamphetamine (basa kuat).