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Reversed Phased Chromatography

・Standard reversed phase packing material for open chromatography
・Enables large-scale preparation and purification at low cost

Particle Size, Flow Rate, and Theoretical Plate Number

Because reversed phase chromatography employs eluents of high viscosity such as methanol and water, the flow rate is lower than that of normal phase chromatography, which uses solvents of low viscosity such as hexane and ethyl acetate. In general, higher theoretical plate numbers can be obtained with lower flow rates.

Packing Material by Column Size

Column size and required amount of C18-PREP packing material

Column I.D. (mm)Bed height (mm)Column volume (ml)Amount of C18-PREP (g)
8 300 15 9
500 25 15
10 300 25 15
500 40 25
20 300 95 55
500 160 95
30 300 210 125
500 350 220
50 300 560 350
500 980 600

Packing Method

  1. Use a standard open glass column, close the stopcock, pack a small amount of absorbent cotton in the bottom of the column and add solvent to approximately 1/3 of the column length.
  2. Add a thin layer (5 mm) of sea sand to the surface of the absorbent cotton.
  3. Prepare a slurry solution of the packing material (30% w/v) with solvent right before packing. (Make sure to prepare enough slurry solution to form a column bed sufficient to separate the compounds of interest.)
  4. Simultaneously open the stopcock and add the slurry solution to the column to form the column bed.
  5. After packing the column, wash the newly packed column bed with 5-10 column volumes of solvent. Allow the bed to stabilize overnight in solvent.
  6. Add a thin layer (5 mm) of sea sand to the top of the bed in order to prevent disturbance of the top of the column bed during sample or solvent addition.

Application Data

Vitamin E Natural Compounds
Basic Compounds
Highly Polar Compounds

Reproducibility and Washing Methods

Wash the COSMOSIL C18-OPN packing material with tetrahydrofuran, chloroform or other solvents to remove the impurities. This packing material has excellent reproducibility and can be used repeatedly.

“CAUTION” Do not wash with basic solvents of pH 7 or more which will dissolve the silica gel or pH 2 or less which will cleave the C18 stationary phase. Dry the packing material at 50°C or less. See end of this chapter for packing method.

Ordering Information

Product Cat.No. PKG Size Price(US$)  
COSMOSIL 50C18-PREP 12065-84 100G 250.00 Buy
COSMOSIL 50C18-PREP 12065-71 1KG 1,800.00 Buy
COSMOSIL 75C18-PREP 12061-24 100G 180.00 Buy
COSMOSIL 75C18-PREP 12061-95 500G 650.00 Buy
COSMOSIL 75C18-PREP 12061-11 1KG 1,000.00 Buy
COSMOSIL 140C18-PREP 12063-04 100G 180.00 Buy
COSMOSIL 140C18-PREP 12063-75 500G 650.00 Buy
COSMOSIL 140C18-PREP 12063-91 1KG 1,000.00 Buy