The concept of the stratography column and its principle of action.

related topics . The type and function of the
column is the main body of gel analysis technology, generally using glass tubes or

glass tubes. Gel
filtration
-layering is the use of a certain size pore with a mesh structure of the gel as a layering medium (such as glucosal gel,
agarose
glue gel, polyacrylamide gel, etc.), according to the molecular size of the separated substance, shape of different diffusion into the gel pores, and therefore through the analysis column of different speed and separation of a layer method.
the column of . The main body of
column is the main body of gel analysis technology, which is generally used with glass tube or coll glass tube. The diameter size of the layer column does not affect the degree of separation, the sample amount is large, can increase the diameter of the column, generally make a backup gel column, diameter is greater than 2 cm, but the sample should be evenly distributed on the surface of the gel column bed. In addition, the diameter increases, the volume of the washed-out liquid increases, and the sample dilution is large. Separation degree depends on column height, in order to separate different components, gel column bed must have a suitable height, separation degree is related to the square root of column height, but due to the soft gel column too high squeeze deformation blocking, generally not more than 1 meter. When the division is separated by short columns, the general gel column is 20-30 cm long, the column height and diameter comparison is 5:1-10:1, and the gel bed volume is 4-10 times the volume of the sample-soluble liquid. The line between column height and diameter is 20:1-100:1 when the grade is separated, and the commonly used gel column is 50×25 cm and 10×25 cm. The dead volume under the stratiline column filter plate should be as small as possible, if the dead volume under the palm filter plate is large, the possibility of remixing between the separated parts is greater, the result is to affect the eloquence of the peak shape, the appearance of drag-tailed image, reduce the dialectic force. At precise separation, the dead volume must not exceed 1/1000 of the total bed volume.to medium pressure
column layer analysis
method to extract separation of winter ling grass meth as an example
medium pressure separation
clind column
operation method to extract separation by medium pressure column layering method For example, the treatment of 1 adsorbents and column filling a certain amount of thin layer of silicone installer enamel plate placed in
oven
, in 105. C after the 2H is active, remove and place at room temperature to load the column. The column is filled with dry method, fills the whole column, does not retain the dead volume, and after sealing, pumps the human solvent according to the ratio of acetone one petroleum ether (5:95) with a plunger-type metering solvent pump until the solvent flows out.2 sample dissolves the ethanol
extract
400G of winter grass with the appropriate amount of acetone, pumps the medium pressure column head, and then pumps the human petroleum ether 1000RID.3to a certain proportion of petroleum ether acetone as an ealing solution, 60ML/MIN flow rate for gradient ealing, column pressure of 2 to 3KG/C. According to each 500ML collection, TLC detection unfolder acetone chloroform (2:8), color agent 5% vanilla aldehyde thick sulfate ethanol liquidto wash away completely, the same fluid merge, recovery solvent. Combine the meth of the winter grass to steam dry. Add a moderate amount of methanol after thermal dissolution of natural placement, crystallization, for the winter grass meth.4 stratospheric column regeneration, balanceactive ingredients by TLC detection of all outflow, with pure acetone 5000ML regeneration, acetone one petroleum ether (5:95) after balance, can be sampled again, repeated use (use times vary with raw materials, generally can be used continuously for more than 2-6 months).5 Recrystrying After the crystallization of the winter herb meth is fully de-stressed, then washed with a small amount of methanol (wash liquid retained for the next hot-soluble winter herb meth). After the winter grass meth is placed with the appropriate amount of methanol heat solution, crystallization is again presumed, to be crystallized completely, filtered, washed with a small amount of ether, will be dried at crystal room temperature after vacuum
drying
boxes, 35 degrees vacuum drying to constant weight. Seal in the bottle and store in the dryer.6 thin layer analysis analysistake the appropriate amount of winter herb meth
control
and the sample methanol from the above method is dissolved on the same silicone G thin sheet, with chloroform acetone (8:2) as the expander, 5% vanilla aldehyde sulfuric acid ethanol liquid color, in 105. C Bake until the spots are clear. It is shown that under this unfolding system, both the sample and control of the winter grass methamphetum have the same RF value and show the same blue-green spots, and there are no impurity marks in the prepared winter grass methicillin samples.
.