Sample loading and efficiency in adsorption, partition and bonded-phase high-speed liquid chromatography
1976; Elsevier BV; Volume: 125; Issue: 1 Linguagem: Inglês
10.1016/s0021-9673(00)93810-x
ISSN1873-3778
Autores Tópico(s)Protein purification and stability
ResumoWith the advent of liquid chromatography columns of extremely high efficiency the effect of sample load on the height equivalent to a theoretical plate (HETP) of the column can be more clearly studied. Previously, the inefficiency of the columns masked the peak broadening due to sample load and deterioration of performance from this cause was only seen to occur at loadings of the order of 100 μg/g of adsorbent. It has now been found that the HETP is a linear function of sample load above about 1 μg/g for microparticulate silica adsorbents which have been slurry packed to give columns with minimum HETP values of between 0.02 and 0.05 mm. The degree of dependence has been found to be a function of both the capacity ratio of the solute and the surface area of the adsorbent. The effect of linear velocity and particle size will be indicated and the relevance of this to the design of preparative liquid chromatography columns will be discussed. Addition of both physically and chemically bonded stationary phases to silica adsorbents is common place in high-speed liquid chromatography and the effect of these changes on the capacity of the columns is an important consideration which will also be outlined.
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