Selected Applications

Hodges Peptide Test Mix

BioAdvantage HL, High Load, is intended for separations requiring maximum loading and selectivity. The high carbon load, 18%, and absence of micro-pores eliminates the problems of other high capacity columns.

Column: BioAdvantage HL C18, 5µm, 4.6 x 150mm
Mobile Phase: A: 0.1% TFA in Water B: 0.1% TFA in Acetonitrile
Gradient: Linear gradient, 1 ml/min, 0 to 30% solvent B in 30 minutes.
Detection: UV 210 nm
Discussion: The Canadian corporation, SPI, produces a well characterized, challenging peptide mixture that is very useful for quantitying HPLC column performance for peptides. The difficult to separate first pair of decapeptides are near base line resolved on Advantage HL C18 columns.
Peak Identities:
1. H2N-RGAGGLGLGK-amide
2. AC-RGGGGLGLGK-amide
3. AC-RGAGGLGLGK-amide
4. AC-RGVGGLGLGK-amide
5. AC-RGWGLGLGK-amide

Hodges Peptides

Tryptic Digest of Transferrin

BioAdvantage 100, a work horse, for years has been the choice for separations of small molecules like carbamate pesticides, pharmaceuticals, synthetic oligonucleotides and small peptides. It is characterized by high efficiency symmetrical peaks and long life

Column: BioAdvantage 100, C18, 5µm, 4.6 x 250mm
Mobile Phase: A: 50 mM phosphoric acid, pH 2.8
B: 60% Acetonitrile / 40% A
Gradient: 1 mi/min
0 min 0%B
5 min 0%B
120 min 40%B
130 min 60%B
Detection: UV 215 nm
Instrument: PerSeptive Biosystems Integral

Tryptic Map of Transferrin

Parabens

Column: BioAdvantage HL, C18, 5µm, 4.6 x 100mm
Mobile Phase: A: 70% Acetonitrile in water
Detection: UV 254 nm
Discussion: Fast analysis, symmetrical peaks and baseline resolution are characteristics of paraben analysis on short, high carbon load C18 columns.
Peak Identities:
1. Methyl paraben
2. Ethyl paraben
3. Propyl paraben
4. Butyl paraben

Parabens

Synthetic Oligonucleotides

Column: BioAdvantage 100, C18, 5µm, 4.6 x 100mm
Mobile Phase: A: 0.1M TEAA, pH 7.0
B: Acetronitrile
Gradient: Linear, 1ml/min from 8-15% ub 24 mins.
Detection: UV 260 nm
Discussion: Trityl-off chromatography is a routine method used for purification and purity determination of synthetic oligonucleotides.

Synthetic Oligonucleotide
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