Sepax Glycomix SAX Columns,Sepax Technologies, Inc._specification/price/image

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Description

Glycomix™ strong anion-exchange phase is made of narrow-dispersed and hydrophilic polymer particles. It is a strong anion exchanger with quaternary ammonium functional groups that are chemically bonded to the hydrophilic surface of the polymer particles.

Packing

Chemical composition of Glycomix™ SAX

Glycomix™ SAX Technical Specifications

Characteristics Glycomix™ SAX

Functional Group Quaternary ammonium

Surface characteristics Hydrophilic

pH range 2 - 12

Typical backpressure for 4.6×250 mm 10 bar for 0.22 mL/min
38 bar for 1 mL/min

Maximum backpressure ~ 1,500 (psi)

Salt concentration range 20 mM - 2.0 M

Maximum temperature ~ 80 (°C)

Mobile phase compatibility Aqueous and organic

Applications Heparin, highly charged polysaccharides

Characteristics	Glycomix™ SAX
Functional Group	Quaternary ammonium
Surface characteristics	Hydrophilic
pH range	2 - 12
Typical backpressure for 4.6×250 mm	10 bar for 0.22 mL/min 38 bar for 1 mL/min
Maximum backpressure	~ 1,500 (psi)
Salt concentration range	20 mM - 2.0 M
Maximum temperature	~ 80 (°C)
Mobile phase compatibility	Aqueous and organic
Applications	Heparin, highly charged polysaccharides

Characteristic

Featured Characteristics

High capacity and high resolution
High lot-to-lot reproducibility
Wide pH range (2-12)
From analytical to preparative scale
Ideal for separation and analysis of heparin, glycans

High Lot to lot Reproducibility

With well-controlled surface chemistry and resin production, Glycomix resins exhibit high lot to lot reproducibility which leads to consistent column performance. The separation variation for Heparin from batch to batch is less than 1% for retention time and 5% for the peak area.

Figure 2. High Lot-to-lot reproducibility of three lots of Glycomix™ SAX resins.

Conditions
Column:	Glycomix™ SAX, 4.6×250mm
Mobile phase:	A: 0.04% NaH₂PO₄, pH 3.0 B: 0.04% NaH₂PO₄+14% NaClO₄, pH 3.0
Flow rate:	0.22 mL/min
Gradient:	20% - 90% B in 60 minutes
Wavelength:	202 nm
Temperature:	25 °C
Injection volume:	10 µL
Pressure:	9.5 bar
Sample:	20 mg/mL Heparin sodium 1 mg/mL Dermatan sulfate (DS) 1 mg/mL Oversulfated chondroitin sulfate (OSCS) in H₂O

Lot-to-Lot peak retention time relative standard deviation.

Peak Name	Lot A	Lot B	Lot C	RSD%
Chondroitin Sulfate B	12.286	12.300	12.434	0.41%
Heparin	21.71	21.723	21.919	0.44%
OSCS	33.936	33.853	34.114	0.32%

Long column life time

Glycomix™ SAX has long column life time due to its resin surface chemistry and stable manufacturing process. Figure below exhibits one column's performance for duplicated injections. The column shows consistent separation efficiency.

Figure 3. Chromatogram overlays of duplicated injections.

Conditions
Column:	Glycomix™ SAX, 4.6×250mm
Guard Column:	Glycomix™ SAX, 4.6×50mm
Mobile phase:	A: 0.04% NaH₂PO₄, pH 3.0 B: 0.04% NaH₂PO₄+14% NaClO₄, pH 3.0
Flow rate:	0.22 mL/min
Gradient:	20% - 90% B in 60 minutes
Wavelength:	202 nm
Temperature:	25 °C
Injection volume:	10 µL
Pressure:	9.5 bar
Sample:	20 mg/mL Heparin sodium 1 mg/mL Dermatan sulfate (DS) 1 mg/mL Oversulfated chondroitin sulfate (OSCS) in H₂O

Sensitive separation of different Heparin loading

Glycomix™ SAX is able to separate components from commercially available standards. In Figure 4, Panel A shows the standard run with 20mg/mL Heparin, 1mg/mL DS and 1 mg/mL OSCS. Panel B is the OSCS chromatogram with 1 mg/mL concentration. In comparison to Panel B, the small peak between DS and Heparin comes from the standard OSCS sample. Column running conditions are the same as for Figure 3.

Glycomix™ SAX can be used in the quantitative analysis of Heparin and its impurities. Figure 5 shows the different heparin loadings over concentration range from 0.5 mg/ml to 10 mg/ml, 0.125 mg/ml to 4 mg/ml for OSCS and 0.25 mg/ml to 4 mg/ml for DS. The R2 values for both standards exceed 0.99 in the tested concentration range.

Figure 4. Chromatogram overlays of OSCS standard and Heparin with DS and OSCS.

Figure 5. Chromatogram overlays of different heparin injections from 0.5mg/mL to 10mg/mL concentration, DS from 0.125 mg/mL to 4.0 mg/mL and OSCS from 0.25 mg/mL to 4.0 mg/mL.

Conditions
Column:	Glycomix™ SAX, 4.6×250mm
Guard Column:	Glycomix™ SAX, 4.6×50mm
Mobile phase:	A: 0.04% NaH₂PO₄, pH 3.0 B: 0.04% NaH₂PO₄+14% NaClO₄, pH 3.0
Flow rate:	0.22 mL/min
Gradient:	20% - 90% B in 60 minutes
Wavelength:	202 nm
Temperature:	25 °C
Injection volume:	10 µL
Pressure:	9.5 bar
Sample:	20 mg/mL Heparin sodium 1 mg/mL Dermatan sulfate (DS) 1 mg/mL Oversulfated chondroitin sulfate (OSCS) in H₂O

Column Performance Comparison

In comparison, Glycomix™ SAX performs superior to Dionex's column for heparin separation.1,2 Figure 7 shows the chromatogram of Heparin separation from Glycomix™ SAX. The resolutions are 3.8 between DS and heparin, 5.8 between heparin and OSCS. The U.S. Pharmacopeia (USP) requirements for the separation for DS/ heparin are 1.0 and 1.5 for OSCS /heparin3. GlycomixTM SAX has a much better separation resolution than USP's requirement. In Dionex's application note, Dionex IonPac AG11 guard with AS11 analytical column gives a 1.1 and 1.8 resolution for DS/heparin and heparin/OSCS respectively.

Figure 7. Heparin Sodium, Chondriotin Sulfate B, and Oversulfated Chondroitin Sulfate separation on Glycomix™ SAX.

Conditions
Column:	Glycomix™ SAX, 4.6×250mm
Guard Column:	Glycomix™ SAX, 4.6×50mm
Mobile phase:	A: 0.04% NaH₂PO₄, pH 3.0 B: 0.04% NaH₂PO₄+14% NaClO₄, pH 3.0
Flow rate:	0.22 mL/min
Gradient:	20% - 90% B in 60 minutes
Wavelength:	202 nm
Temperature:	25 °C
Injection volume:	10 µL
Pressure:	10 bar
Sample:	20 mg/mL Heparin Sodium 0.2 mg/mL Dermatan Sulfate (DS) 0.2 mg/mL Oversulfated Chondroitin Sulfate (OSCS)