HPLC

Discovery BIO Wide Pore C5

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Ordering Information

 
Discovery BIO Wide Pore C5
HPLC Columns and Capillaries
 
Bonded Phase:   Covalently-bonded pentylsilane (C5), endcapped
Silica:   Spherical, 3µm, 5µm and 10µm diameters, 300Å pore diameter,
    high purity (<10ppm metals)
 
Solutions for Protein and Peptide Separation Challenges
 

Proteomics and biotherapeutics: high efficiency protein and hydrophobic separations

LC/MS; no-bleed, capillary dimensions, and pH stability

Protein purification: Scalable from analytical to preparative

Enhanced lifetime and pH stability over C4 phases


 
Here's the
Proof
 
EXCELLENT PERFORMANCE FOR LARGE MOLECULES
 

Column: Discovery BIO Wide Pore C5, 15cm x 4.6mm, 5µm; Mobile Phase: (A) 0.1% (v/v) TFA in H2O: MeCN (75:25), (B) 0.1% (v/v) TFA in H2O: MeCN (25:75); Flow Rate: 1mL/min; Temp: 30°C; Detection: 220nm; Injection: 10µL; Sample: Protein Test Mix; Gradient: 0-100% B in 25 min.

 
1. RNase (1mg/mL) (13.7kDa)
2. Cytochrome C (1mg/mL) (12.4 kDa)
3. Lysozyme (1mg/mL) (14.3 kDa)
4. Bovine Serum Albumin (2.5mg/mL) (67.0 kDa)
5. Myoglobin (1mg/mL) (18.8 kDa)
6. Ovalbumin (3.5mg/mL) (45.3 kDa)
 

g001488_1533


 
INCREASED STABILITY OF DISCOVERY BIO WIDE PORE C5 OVER CONVENTIONAL C4
 

After 25,000 column volumes (222 gradient cycles) were passed through the column, the Discovery BIO Wide Pore C5 showed very little change in efficiency. The competitive C4 column showed a significant loss of efficiency, as much as 50%.

 

Column: Discovery BIO Wide Pore C5, 5cm x 4.6mm, 5µm; Mobile Phase: (A) 0.1% TFA in H2O : MeCN (95:5), (B) 0.1% TFA in H2O : MeCN (25:75); Flow Rate: 2mL/min; Temp: 35°C; Detection: 220nm; Injection: 5µL; Sample: Protein/Peptide Test Mix; Gradient: 0-20% B in 20 min

 
g001588

 
STABILITY OF DISCOVERY BIO WIDE PORE C5 AT pH 8.0
 

Retention times of the sample components remained constant after 14,000 bed volumes were passed through the column.

 

Column: Discovery BIO Wide Pore C5, 5cm x 4.6mm, 5µm; Mobile Phase: 25mM potassium phosphate, pH 8 : MeOH (95:5); Flow rate: 2mL/min; Temp: 35°C; Detection: 254nm

g001491


 
UNDETECTABLE LC/MS BLEED
 

LC/MS Bleed Analysis on Discovery BIO Wide Pore C5
Column: Discovery BIO Wide Pore C5, 15cm x 4.6mm, 3µm; Mobile Phase: (A) 0.1% TFA in H2O, (B) 0.1% TFA in MeOH; Flow Rate: 1mL/min; Temp: 30°C; Gradient: 0-100% B in 15 min, hold for 5 min, then back to 0% B for 10 min

 

No TIC baseline rise relative to blank run (gradient run w/o column)

g001495

 

M/S Spectrum (shaded area above)

g001496

 

Essentially no m/z peaks generated from column when compared to blank run.
("X" indicates peaks also found in blank run)


 
LOT TO LOT REPRODUCIBILITY OF DISCOVERY BIO WIDE PORE C5
 

Column: Discovery BIO Wide Pore C5, 15cm x 4.6mm, 5µm; Mobile Phase:(A) 0.1% PFPA (pentaflouropropionic acid) in H2O : MeCN (81:19), (B) 0.1% PFPA in H2O : MeCN (62:38); Flow Rate: 1mL/min; Temp: 30°C; Detection: 215nm; Injection: 10µL; Sample: Peptide Mix (Sigma Cat. No. P2693); Gradient: 0-100% B in 19 min.

 

Peak Identity:

Amino Acid Sequence:

1. Arg8-vassopressin
2. Brandykinin, fragment 1-5
3. Oxytocin
4. Met-enkaphalin
5. Luteinizing Hormone Releasing Hormone
6. Leu-enkephalin
7. Brandykinin
8. Bombesin
9. Substance P

CFQNCPRG-amide; disulfide
RPPGF
CYIQNCPLG-amide; disulfide
PEHWSYGLRPG-amide
YGGFM
YGGFL
RPPGFSPFR
PEQRLGNQWAVGHLM-amide
RPKPQQFFGLM-amide

g001492_94


 

Ordering Information
3µm Discovery BIO Wide Pore C5 HPLC Columns
ID(mm) Length(cm) Cat. No.
1.0 10 65511-U
2.1 5 567226-U
2.1 10 567227-U
2.1 15 567228-U
4.6 5 567229-U
4.6 10 567230-U
4.6 15 567231-U
5µm Discovery BIO Wide Pore C5 HPLC Columns
ID(mm) Length(cm) Cat. No.
2.1 5 568400-U
2.1 15 568402-U
4.6 5 568420-U
4.6 10 568421-U
4.6 15 568422-U
4.6 25 568423-U
10.0 25 568430-U
10µm Discovery BIO Wide Pore C5 HPLC Columns
ID(mm) Length(cm) Cat. No.
10.0 15 567234-U
10.0 25 567235-U
2cm Supelguard Cartridges with Discovery BIO C5 Packings
Description 2.1mm ID 4.0mm ID
3µm Particle Size
2 Pack 567278-U 567280-U
Kit 567279-U 567281-U
5µm Particle Size
2 Pack 568470-U 568472-U
Kit 568471-U 568473-U
  For 4.0mm ID or 4.6mm ID analytical columns
triangle Kit includes a holder and one replaceable cartridge.

 
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USA only:
Phone: 800-247-6628
  or 814-359-3441
fax: 800-325-5052
email: techservice@sial.com
 
Outside the USA: click here

 
TRADEMARKS
Discovery and Supelguard are trademarks of Sigma-Aldrich Co.