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.
	Detector	220 nm
	Column	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
	Gradient	25 to 62 % B in 2 min (9.6 Column Volume)
	Flow rate	4 ml/min (1,450 cm/hr)
	Temperature	30°C
	Injection volume	2.5  µl
	Sample: (1 mg/ml in buffer A)	Ribonuclease A, Cytochrome c, Lysozyme, BSA, Myoglobin, OVA

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.
	Detector	220 nm
	Column	STYROS™ 3R/XH 4.6 X 250 mm
	Mobile Phase	A: 10 mM Phosphate, pH=11.2 B: ACN
	Gradient	5 to 15 % B in 30 min (7.2 Column Volume)
	Flow rate	1 ml/min (360 cm/hr)
	Temperature	30°C
	Injection volume	100  µl
	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.
	Detector	220 nm
	Column	STYROS™ 3R/XH 4.6 X 100 mm
	Mobile Phase	A: 10 mM Phosphate, pH=1.5 B: ACN
	Separation	Isocratic: 15 % B
	Flow rate	1.5 ml/min (540 cm/hr of linear flow rate)
	Temperature	30°C
	Injection volume	0.5  µl
	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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