Chromatography - 4. Packing of a Liquid Chromatograph (LC) Separation Column

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Chromatography - 4. Packing of a Liquid Chromatograph (LC) Separation Column
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Chromatographie - 4. Packen einer flüssigkeitschromatographischen Trennsäule
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C 1589
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Film, 16 mm, LT, 52 ; F, 5 min

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In der Niederdruck-Flüssigkeitschromatographie werden handelsübliche oder selbstgepackte Trennsäulen benutzt. Deren Trennleistung hängt von der Qualität der Säulenpackung ab. Demonstration einzelner Fertigungsschritte: Einfüllen von Kieselgel und Lösungsmittel; Verdichtung der Packung; Test mit Farbstoffmischung; Zonenbildung als Qualitätstest.
Each of the steps involved in the homogenous packing of a separation column with silica gel as a stationary phase for low pressure-hypotension liqiuid chromatography is described. A dye mix is used to test the quality, or efficiency, of separation of the column. This is done by comparing the column with columns which have already been completely separated.
Keywords Chromatographie Trennsäule Testgemisch Säulenherstellung Qualitätstest Niederdruck Lösungsmittelzugabe Kieselgel Homogenisierung Flüssigkeitschromatographie Füllmaterial Farbstofflösung dye solution packing material liquid chromatography homogenization silica gel addition of solvent low pressure quality control production of columns test mix separation column chromatography
Stop codon Separation process Zigarettenschachtel Glasses Column chromatography
Homogeneous (chemistry) Separation process Drop (liquid) Zigarettenschachtel Silicon dioxide Chromatography Dörren Silicone Thermoforming Stuffing
Homogeneous (chemistry) Stop codon Separation process Zigarettenschachtel Potato chip Thin film Legieren Nanoparticle Cork taint
Separation process Zigarettenschachtel Glasses Pigment Mixture
Sample (material) Potato chip Silicon dioxide Tube (container)
Sample (material) Separation process Desiccant Silicon dioxide Elution Reservoir Deformity
Separation process Elution
Homogeneous (chemistry) Volumetric flow rate Zigarettenschachtel Elution Nitrosamine
The separation efficiency of a liquid chromatographic column essentially depends on the quality of its packing. Between two commercial columns an empty glass valve for the preparation of the separation column for low pressure liquid chromatography, for short LC. A
small roll of cotton is pressed tightly with a glass rod into the inlet of solvent the air is removed. a stop cork. After moistening
Dry separation material, that is silicon gel, is stirred with with a few drops of an organic solvent and later forms the stationary phase. A
funnel serves for easy filling of the separation column. The stationary phase gradually forms a homogeneous packing of silica gel
particles. An excess of solvent is drained off with only a few millimetres thick layer left over. The roll of cotton just above the stop cork retains the separation particles. Finally the required height of the separation column has been attained. Knocking with cork rings condenses the packing of the column and improves its homogeneity.
A thin chip of glass fibre fleece will later protect
the uppermost layer of the column from being whirled up. The enclosed air is carefully removed with a glass rod.
The packing efficiency of the column is now tested with a pigment mixture. An optimal separation can be achieved only if the pigment mixture is introduced as a homogeneously concentrated
layer on top of the glass fibre chip at the starting point of the chromatographic column. The sample will be most conveniently allowed to run down around the inner wall of the open tube. Shortly afterwards the coloured substances soak into the silica gel. The
introduced sample is carefully superimposed by the mobile phase. Only by this procedure can the coloured zone be prevented from
deformation already at the start of the separation. The mobile phase must be filled up quickly to prevent desiccation of the uppermost silica gel layer. Finally, a big reservoir containing
mobile phase is placed on top of the column. During
the following separation process the solvent continuously runs down speeded up her about 70 times.
The performances of the three different columns depend on the homogeneity of the packing and flow of the mobile phase.


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