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Proto 300 C18 Proto 300 C4

Applications

Low pH operating conditions and high pH column washing steps are typical bioprocessing operating conditions that can severely limit the lifetime and performance of widepore columns typically used for protein and peptide analysis. Higgins Analytical’s new PROTO300 has high capacity for enhanced resolution and is robust enough to withstand prolonged use at extreme pH (1.5 - 10).

Stability Study Under Extremely

Alkaline Conditions

Experimental: 150 x 4.6mm columns were studied. All experiments were conducted on duplicate columns. Solutes used for performance measurement were dimethylphthalate and fluorene.

Step One:

Column stability and measurement precision was established for each column with multiple injections over 18hrs under neutral conditions (70% MeCN/water, 1mL/min). All columns were very stable. The chromatograms of a column from each manufacturer at the end of the 18 hour period are shown on the left hand side.

Step Two:

Each column was flushed with 70% MeCN/0.5N NaOH solution at 1mL/min for one hour (23ºC). The columns were then purged and equilibrated under the standard neutral conditions then retested. The center chromatograms are the results of the first test after neutral equilibration

Step Three:

After the second step, the neutral test conditions were maintained for one hour and a repeat injection was made. The results are illustrated in the chromatograms on the right hand side of the figure. While Higgins Analytical does not recommend flushing any silica-based HPLC column under such aggressive conditions, these data show the superior akaline stability of Proto™ 300 C18.

PROTO300 Robust Bioseparations

Phases C4 and C18

Particle Sizes 4.5 and 10μm

Pore Size 300Å

Pore Volume 0.9mL/gm

Surface Area 100m

2/gm

%Carbon (w/w) C4 = 3%, C18= 8%

Phase type Monofunctional & fully endcapped

Silica Class Type B

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Proto 300 C4 5um 1.0 x 10.0mm Piccolo Column Proto 300 C18 5um 1.0 x 10.0mm Piccolo Column Proto 300 C4 5um 0.3 x 2.5mm Piccolo Column
RP-0101-W045 RP-0101-W185 RP-M2M3-W045
Proto 300 C18 5um 0.3 x 2.5mm Piccolo Column Proto 300 C4 5um 0.5 x 2.5mm Piccolo Column Proto 300 C18 5um 0.5 x 2.5mm Piccolo Column
RP-M2M3-W185 RP-M2M5-W045 RP-M2M5-W185
Proto 300 C4 5um 1.0 x 5.0mm Piccolo Column Proto 300 C18 5um 1.0 x 5.0mm Piccolo Column Proto 300 C4 5um 2.1 x 10mm Sprite Column
RP-M501-W045 RP-M501-W185 RR-0121-W045
Proto 300 C18 5um 2.1 x 10mm Sprite Column Proto 300 C4 5um 2.1 x 20mm Sprite Column Proto 300 C18 5um 2.1 x 20mm Sprite Column
RR-0121-W185 RR-0221-W045 RR-0221-W185
Proto 300 C4 5um 2.1 x 40mm Sprite Column Proto 300 C18 5um 2.1 x 40mm Sprite Column Proto 300 C4 5um 1.0 x 30.0mm Piccolo Column
RR-0421-W045 RR-0421-W185 RP-0301-W045
Proto 300 C18 5um 1.0 x 30.0mm Piccolo Column Proto 300 C4 5um 75um and 150um 0.075 x 250mm Capillary Column Proto 300 C18 5um 75um and 150um 0.075 x 250mm Capillary Column
RP-0301-W185 RC-2507-W045 RC-2507-W185
Proto 300 C4 5um 75um and 150um 0.150 x 250mm Capillary Column Proto 300 C18 5um 75um and 150um 0.150 x 250mm Capillary Column Proto 300 C4 10um 10 x 50mm Semi Prep Column
RC-2515-W045 RC-2515-W185 RS-0510-W041
Proto 300 C18 10um 10 x 50mm Semi Prep Column Proto 300 C4 5um 2.1 x 30mm Analytical Column Proto 300 C18 5um 2.1 x 30mm Analytical Column
RS-0510-W181 RS-0321-W045 RS-0321-W185
Proto 300 C4 5um 3.0 x 30mm Analytical Column Proto 300 C18 5um 3.0 x 30mm Analytical Column Proto 300 C4 5um 4.0 x 30mm Analytical Column
RS-0330-W045 RS-0330-W185 RS-0340-W045
Proto 300 C18 5um 4.0 x 30mm Analytical Column Proto 300 C4 5um 4.6 x 30mm Analytical Column Proto 300 C18 5um 4.6 x 30mm Analytical Column
RS-0340-W185 RS-0346-W045 RS-0346-W185
   
 
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