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G1006458

HPLC Analysis of Protein Standards on Zenix® SEC-300 by Size Exclusion As Affected by Arginine

application for HPLC

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technique(s)

HPLC: suitable

test parameters

column: Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 μm particles (A777028)
mobile phase: [A], 0.15 M potassium phosphate, monobasic, pH 7.0 with potassium hydroxide; [B] 0.15 M potassium phosphate, monobasic, 0.2 M arginine, pH 7.0 with phosphoric acid (mobile phase is either 100% A or 100% B)
flow rate: 0.3 mL/min
pressure: 1030 psi 100% A (71 bar)
pressure: 1165 psi 100% B (80 bar)
column temp.: 25 °C
detector: UV, 280 nm
injection: 2.5 μL
sample: Waters BEH200 SEC protein standard

suitability

application for HPLC

application(s)

life science and biopharma

Analysis Note

Arginine can have benefitial effects on peak shape and recovery of hydrophobic proteins when chromatographed by size exclusion chromatography. In this case, with protein standards that are not particularly hydrophobic, no dramatic affect is noted, as assessed by peak efficiency, peak widths, or peak retention.

Legal Information

Zenix is a registered trademark of Sepax Technologies

Analyte

Description

    Thyroglobulin from bovine thyroid, (For Gel Filtration Chromatography)
    IgG from rabbit serum, reagent grade, ≥95% (SDS-PAGE), essentially salt-free, lyophilized powder
    Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
    Myoglobin from equine heart, ≥90% (SDS-PAGE), essentially salt-free, lyophilized powder
    Uracil, ≥99.0%

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