PubMed · 2417282
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N Shimamoto. 1985. [Multi-mixing technique].. https://pubmed.ncbi.nlm.nih.gov/2417282/
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In this paper the key steps towards drug screening in biological fluids are considered: (i) sample work up-isolation-concentration; (ii) differentiation-detection; (iii) identification. For (i) solid-phase extraction has very good potential; for (ii) thin-layer chromatography, gas chromatography and high-performance liquid chromatography, in combination with their respective detection modes, each have their particular advantages and disadvantages; for (iii) extensive computerized data bases are essential. It is emphasized that each step must be approached and carried out in a systematic way, not only to make sure that all substances of interest can be detected, but also for their correct identification. To this end all analytical procedures and techniques used require extensive validation of their detection and identification properties.
A novel chiral stationary phase that contains a new chiral selector based on L-valine-3,5-dimethylanilide attached to monodisperse poly(glycidyl methacrylate-co-ethylene dimethacrylate) beads has been prepared. The polymeric separation medium provides greatly enhanced enantioselectivities and reduced retention times when compared to the analogous silica-based chiral stationary phase in the separation of the enantiomers of 3,5-dinitrobenzamido derivatives of alpha-amino acids under normal-phase HPLC conditions. Separation factors (alpha) of up to 7 were achieved with the polymeric separation medium, which is also useful for the separation of large samples under overload conditions.
Blue native polyacrylamide gel electrophoresis (BN-PAGE) or colorless native polyacrylamide gel electrophoresis (CN-PAGE) allowed separation of the oxidative phosphorylation complexes of yeast mitochondria. These complexes were characterized by specific staining related to their enzymatic activity. Solubilization of mitochondria by different nonionic detergents such as Triton X-100, dodecyl maltoside, Nonidet P-40, Lubrol, octyl glucoside, or Hecameg led to the separation of F1-FO ATPase complexes exhibiting distinct apparent molecular masses related to different contaminating proteins and lipids. All these different forms were active in ATP hydrolysis as revealed directly on the gel. Analysis of the subunit composition of these complexes was carried out by a two-dimensional Tricine-SDS-PAGE and showed that the purest F1-FO ATPase complex was obtained with Lubrol, whereas with Hecameg and octyl glucoside, only the F1 part of ATPase was solubilized.