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APPLICATION NOTE

AN030108-49

STYROS™ 3R Simulated Monolith Polymeric Reversed Phase.

 

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 Simulated Monolith™ in the separation of biomolecules as well as smaller peptides and other molecules under different and extreme conditions.

 

Chromatogram 1

Separation of 6 proteins on STYROS™ 3R/XH
(Flow Rate: 4 ml/min)

Table 1. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 3R/XH 4.6 X 50 mm

Mobile phase.

A: 0.075 % TFA in H2O
B: 0.075 % TFA in ACN:H2O 95:5

Flow rate

4 ml/min (1,450 cm/hr )

Gradient

25 to 62 % B in 2 min (9.6 cv).

Temperature

30°C

Detection

220 nm

Injection volume

2.5 ul

Sample:

Ribonuclease A, Cytochrome c, Lysozyme, BSA, Myoglobin, OvA
(1 mg/ml in buffer A)


 

Chromatogram 2

Separation of 7 peptides on STYROS™ 3R/XH
(Flow Rate 1 ml/min)

Table 2. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 3R/XH 4.6 X 250 mm

Mobile phase.

A: 10 mM Phosphate, pH=11.2
B: ACN

Flow rate

1 ml/min (360 cm/hr )

Gradient

5 to 15 % B in 30 min (7.2 cv).

Temperature

30°C

Detection

220 nm

Injection volume

100 ul

Sample:

1. Gly-Tyr, 2. Val-Tyr-Val, 3. Met enkephalin, 4. Leu enkephalin,
5. Angiotensin II, 6. Angiotensin III, 7. Angiotensin I
(0.5 mg/ml each in DI H2O)

 

Chromatogram 3

Separation of Benzoic acid and its derivatives on STYROS™ 3R/XH
(Flow Rate 1.5 ml/min)

Table 3. Operating parameters.

HPLC System.

Agilent 1100 with thermostatted column compartment.

Columns

STYROS™ 3R/XH 4.6 X 100 mm

Mobile phase.

A: 10 mM Phosphate, pH=1.5
B: ACN

Flow rate

1.5 ml/min (540 cm/hr of linear flow rate)

Gradient

Isocratic: 15 % B

Temperature

30°C

Detection

230 nm

Injection volume

0.5 ul

Sample:

1. 3,5-Dihydroxybenzoic acid, 2. 4-Hydroxybenzoic acid,
3. 2,4-Dihydroxybenzoic acid, 4. Acetyl Salicylic acid, 5. Benzoic acid,
6. Salicylic acid, (170 ug/ml each in A:B 50:50)

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

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