![Page 1: Method of Chromatography prof. aza Department of Pharmacy, Andalas University STIFI Perintis STIFAR Pekan Baru STIFI Bhakti Pertiwi](https://reader035.vdocuments.site/reader035/viewer/2022062305/5697bfd81a28abf838caeff1/html5/thumbnails/1.jpg)
Method of Chromatography
prof. azaDepartment of Pharmacy, Andalas University
STIFI Perintis
STIFAR Pekan Baru
STIFI Bhakti Pertiwi
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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
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Characteristic of a Gaussian band
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Band asymmetry, (a) Definition of asymmetry factor, As; (b-e) examples of band asymmetry
![Page 5: Method of Chromatography prof. aza Department of Pharmacy, Andalas University STIFI Perintis STIFAR Pekan Baru STIFI Bhakti Pertiwi](https://reader035.vdocuments.site/reader035/viewer/2022062305/5697bfd81a28abf838caeff1/html5/thumbnails/5.jpg)
Separation of two bands as a function of resolution (Rs) and relative band size (I/1, ¼, 1/16).
![Page 6: Method of Chromatography prof. aza Department of Pharmacy, Andalas University STIFI Perintis STIFAR Pekan Baru STIFI Bhakti Pertiwi](https://reader035.vdocuments.site/reader035/viewer/2022062305/5697bfd81a28abf838caeff1/html5/thumbnails/6.jpg)
The basic measurement
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Effects of a change in k’, N, or a on the resolution of two bands
![Page 8: Method of Chromatography prof. aza Department of Pharmacy, Andalas University STIFI Perintis STIFAR Pekan Baru STIFI Bhakti Pertiwi](https://reader035.vdocuments.site/reader035/viewer/2022062305/5697bfd81a28abf838caeff1/html5/thumbnails/8.jpg)
Peak resolution
R = 2d/(w1 + w2)
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Relationship of mean retention time (tmean) and observed (mode) retention time (tR to EMG time constant (t)
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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,
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Linear dependence of log k’ on reciprocal temperature.
GC separation on Apiezon L
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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
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Dependence of retention on pH in the reversed-phase LC separation of various acidic, neutral, and basic compound
Separation of several bile acids
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Separation of (1) salicylic acid (strong acid), (2) phenobarbital (weak acid), (3) phenacetin (neutral), (4) nicotine (weak base), (5)methylamphetamine (basa kuat).