OraChrom, Inc.

  
       

 

 

Home
Up

APPLICATION NOTE

AN030308-50

STYROS™ 3R Simulated Monolith Polymeric Reversed Phase: Standard Separation of 6 Small Molecules

 

Reversed phase chromatography with silica provides high resolving power but very limited stability due to the solubility of the matrix. The separations are therefore limited to neutral pH’s for a limited number of runs.

Polymeric on the other hand have been used successfully to alleviate the issues of chemical instability associated with silica matrices and therefore have provided the possibility of using the full pH range for baseline separations.

The following chromatogram shows the separation of 6 small molecules on a 3R Simulated Monolith reversed phase column.

 

Chromatogram 1

Separation of 6 small molecules on STYROS™ 3R/XH
(Flow Rate: 2 ml/min on normal bore column at 30˚ C)

 

Table 1. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 3R/XH 4.6 X 50 mm (V=0.83 ml)

Mobile phase.

A: H2O
B: ACN

Flow rate

2 ml/min (720 cm/hr of linear flow rate )

Gradient

Isocratic: 40 % B

Temperature

30°C

Detection

254 nm

Injection volume

10 ul

Sample:

1. Thiourea, 2. Benzyl alcohol, 3. Aniline, 4. Nitrobenzene, 5. Methyl Salicylate, 6. Toluene (30-300 ug/ml each in A:B 50:50)

 

Full separation can be achieved on a short 5 cm column at linear flow rates of 700 cm/hr. 

Increasing the column length to 10 cm and raising the temperature to 60˚ C the separation can be improved further even on a narrow bore column of 2.1mm ID and linear flow rates of 1,200 cm/hr.

 

 
Chromatogram 2

Separation of 6 small molecules on STYROS™ 3R/XH
(Flow Rate 0.7 ml/min on narrow bore column)

Table 2. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 3R/NB 2.1 X 100 mm

Mobile phase.

A: H2O
B: ACN

Flow rate

0.7 ml/min (1,200 cm/hr of linear flow rate)

Gradient

Isocratic: 25 % B

Temperature

60°C

Detection

254 nm

Injection volume

2.5 ul

Sample:

1. Thiourea, 2. Benzyl alcohol, 3. Aniline, 4. Nitrobenzene, 5. Methyl Salicylate, 6. Toluene (30-300 ug/ml each in A:B 50:50)

 

Hard gel polymeric gigaporous stationary phases offer similar mechanical stability compared to silica and far superior chemical stability regarding extremes of pH's.

The present application note highlights the capabilities of STYROS™ 3R.

Unlike Monolith STYROS™ Simulated Monolith columns are available in many sizes for additional resolving capabilities.

[ Home ] [ Up ]

Copyright © 1997-2009 OraChrom, Inc.