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| Discovery BIO Wide Pore C5 |
| HPLC Columns and Capillaries |
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| Bonded Phase: |
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Covalently-bonded pentylsilane (C5), endcapped |
| Silica: |
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Spherical, 3µm, 5µm and 10µm diameters, 300Å pore diameter, |
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high purity (<10ppm metals) |
| Column IDs: |
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0.18mm to 21.2mm |
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| Solutions for Protein and Peptide Separation Challenges |
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Proteomics and biotherapeutics: high efficiency protein and hydrophobic separations
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LC/MS; no-bleed, capillary dimensions, and pH stability
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Protein purification: Scalable from analytical to preparative
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Enhanced lifetime and pH stability over C4 phases
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| EXCELLENT PERFORMANCE FOR LARGE MOLECULES |
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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.
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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) |
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| INCREASED STABILITY OF DISCOVERY BIO WIDE PORE C5 OVER CONVENTIONAL C4 |
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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%.
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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
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| STABILITY OF DISCOVERY BIO WIDE PORE C5 AT pH 8.0 |
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Retention times of the sample components remained constant after 14,000 bed volumes were passed through the column.
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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
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| UNDETECTABLE LC/MS BLEED |
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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
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No TIC baseline rise relative to blank run (gradient run w/o column)
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M/S Spectrum (shaded area above)
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Essentially no m/z peaks generated from column when compared to blank run.
("X" indicates peaks also found in blank run)
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| LOT TO LOT REPRODUCIBILITY OF DISCOVERY BIO WIDE PORE C5 |
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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.
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Peak Identity:
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Amino Acid Sequence:
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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
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CFQNCPRG-amide; disulfide
RPPGF
CYIQNCPLG-amide; disulfide
PEHWSYGLRPG-amide
YGGFM
YGGFL
RPPGFSPFR
PEQRLGNQWAVGHLM-amide
RPKPQQFFGLM-amide
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