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Discovery® HPLC Columns for Fast LC-MS Analysis

LC-MS Columns

The Discovery LC-MS line offers short length columns for fast LC-MS analysis that are rugged and dependable. Supelco offers a robust product that withstands column shock due to fast gradient turnaround times. Columns installed on LC-MS instruments are typically operated using gradients of 5%-95% organic with gradient cycle times of 2-3 minutes. Our column lifetime is maximized through the use of stable bonding chemistry, high purity silica and optimal proprietary packing procedures.

The Discovery line of HPLC columns offers excellent retentivity, selectivity and reproducibility for optimal results. LC-MS formats are now available with Discovery C18, C8, Cyano and HS F5 phases.

Discovery LC-MS Columns Deliver:

Properties C18 C8
Bonded Phase: Octadecylsilane, endcapped Octylsilane, endcapped
Silica: Spherical, high purity, <10 ppm metals Spherical, high purity, <10 ppm metals
Particle Size: 5 µM 5 µM
Pore Size: 180 Å 180 Å
Surface Area: 200 m2/g 200 m2/g
%C: 12% 7.5%
Coverage: 3 µmol/m2 3.4 µmol/m2
C18 structure C8 structure
Properties CYANO HS F5
Bonded Phase: Cyanopropylsilane, endcapped Pentafluorophenylpropyl, endcapped
Silica: Spherical, high purity, <10 ppm metals Spherical, high purity, <10 ppm metals
Particle Size: 5 µM 3.5 µM
Pore Size: 180Å 120Å
Surface Area: 200 m2/g 300 m2/g
%C: 4.5% 12%
Coverage: 3.5 µmol/m2 4 µmol/m2
Cyano structure HS F5 structure

High Speed Separations

Discovery LC-MS columns can shorten analysis times to less than 4 minutes for the study of antibiotics.

Figure 1. Rifampicin on Discovery C18

Column 1: Discovery C18, 15 cm x 2.1 mm ID, 5 µM particles
Cat. No.: 50495521
Column 2: Discovery C18, 2 cm x 2.1 mm ID, 5 µM particles
Cat. No.: 577507-U
Mobile Phase: 60:40, 33 mM HCO2H:CH3CN
Flow Rate: 0.2 mL/min
Temp.: 35 ºC
Det.: ESI(+), full scan
Inj.1: 6 µL
Inj.2: 0.8 µL
Sample: 10 mg/L in 50% CH3OH
Discovery C18
Equation
Discovery C18
Figure 2. Application of Short, Fast LC-MS Columns for Analysis of Veterinary Antibiotics on Discovery C18 and C8 Columns
Column 1: Discovery C18, 2 cm x 2.1 mm ID,
5 µM particles
Cat. No.: 577507-U
Column 2: Discovery C8, 2 cm x 2.1 mm ID,
5 µM particles
Cat. No.: 577501-U
Mobile Phase: A=50mM HOAc/NH4OH, (pH 6.0), B=water, C=CH3OH
Flow Rate: 0.2 mL/min
Temp.: 65 ºC
Inj.: 1 µL
Det.: ESI(-)
Sample:
1. Lasalocid A (1 µg/mL)
2.3. Salinomycin (10 µg/mL)
4. Narasin (5 µg/mL)
Discovery C18, A:B:C = 20:5:75 Discovery C8, A:B:C = 20:10:70
Discovery C18
Discovery C8
Column 1: Discovery C18, 2 cm x 2.1 mm ID, 5 µM particles
Cat. No.: 577507-U
Column 2: Discovery C8, 2 cm x 2.1 mm ID, 5 µM particles
Cat. No.: 577501-U
Column 3: Discovery Cyano, 3 cm x 2.1 mm ID, 5 µM particles
Cat. No.: 577514-U
Mobile Phase: A=50 mM HCO2H, B=water, C=90:10 CH3OH:CH3CN
Flow Rate: 0.2 mL/min
Temp.: 35 ºC
Inj.: 1 µL
Det.: ESI(+)
Sample:
1. Erythromycin c (1 µg/mL)
2. Clarithromycin (1 µg/mL)

Discovery C18
A:B:C = 20:35:45

Discovery C18

Discovery C8
A:B:C = 20:35:45

Discovery C8

Discovery Cyano
A:B:C = 20:40:40

Discovery CYANO

Mechanical Stability via Fast Gradient Testing

The Supelco Discovery LC-MS HPLC columns are rigorously tested in order to provide the highest mechanical stability under repeated gradient cycles. Table 1 shows of the impact on column efficiency, peak shape, retention times and back pressure after 1000 cycles of a 2-minute rapid (ballistic) gradient with 1-minute equilibration period.

There is no loss of column efficiency or increase in tailing or column backpressure, and no change in retention time. See Table 2 for details of testing.

PhaseEfficiency
(N/column)
USP Tailing
Factor
Retention Time
(min.)
Pressure
(bar)
Discovery C18 (3 cm x 3.0 mm ID)
Before stability test27921.041.98939
After stability test28991.091.98541
Discovery C8 (2 cm x 2.1 mm ID)
Before stability test14511.051.76726
After stability test15071.091.76125
Discovery Cyano (3 cm x 4.6 mm ID)
Before stability test33291.011.36844
After stability test34481.011.36240
Table 1.Rapid Gradient Turnaround Testing of Discovery LC-MS HPLC Columns
Discovery C18 (3 cm x 3.0 mm ID)
Flow: 0.8 mL/min
Gradient: 10-90% methanol over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.: 30 °C
Wavelength: 254 nm
Pressure: 76 bar for first cycle
75 bar after last cycle
No. of Cycles: 1,135 three-minute
Total Volume: 2724 mL
Void Volume: 0.447
Discovery C8 (2 cm x 2.1 mm ID)
Flow: 0.5 mL/min
Gradient: 10-90% methanol over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.: 30 °C
Wavelength: 254 nm
Pressure: 40 bar after first cycle
38 bar after last cycle
No. of Cycles: 1,089 three-minute
Total Volume: 1634 mL
Void Volume: 0.393
Discovery Cyano (3 cm x 4.6 mm ID)
Flow: 0.2 mL/min
Gradient: 10-50% acetnonitrile over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.: 30 °C
Wavelength: 254 nm
Pressure: 72 bar after first cycle
70 bar after last cycle
No. of Cycles: 937 three-minute
Total Volume: 5622 mL
Void Volume: 0.434
Table 2.Test Conditions

Discovery C18 Bleed Analysis

Mass spectral responses from background ions often inhibit both qualitative and quantitative analysis in LC-MS experiments. Common sources of background ions include the solvents, interface or system contamination, and HPLC column bleed. The presence of background ions can be reduced by using high quality solvents such as LC-MS, maintaining a clean LC-MS system, and by using high quality HPLC stationary phases from Supelco.

Supelco utilizes technologically advanced bonding chemistry and highly purified silica to minimize the potential for column bleed.

LC-MS Bleed Analysis

The Total Ion Chromatogram (TIC) shown in the top of Figure 4 shows the result from an MS detector of one gradient cycle with a Discovery C18, 2 cm x 2.0 mm ID column in-line. The two selective ion chromatograms (SIC) in the bottom of Figure 4 show the mass spectrum taken from the shaded region of the TIC (at the high organic content point in the gradient). The bottom right SIC shows that the Discovery C18 column does not introduce any additional masses that were not in the blank run (bottom left SIC). The Discovery C18 column exhibits low bleed for LC-MS analyses.

Total Ion Chromatogram

Discovery C18, 2 cm x 2.0 mm ID, acetonitrile gradient in presence of 0.1% formic acid / 5 cycles / ESI (+) mode acquisition

LC-MS Bleed Study

Figure 4.LC-MS Bleed Study.

Blank Run

Blank Run (no LC column) (19-21 min.)

Discovery C18 Column

Discovery C18 column (19-21 min.)

Ordering Information

PhaseID (mm)Length (cm)Product. No.
5 µM Discovery C8 Columns
2.12577501-U
2.13577502-U
3.02577503-U
3.03577504-U
4.62577505-U
4.63577506-U
5 µM Discovery C18 Columns
2.12577507-U
2.13577508-U
3.02577509-U
3.03577510-U
4.62577511-U
4.63577512-U
5 µM Discovery Cyano Columns
2.12577513-U
2.13577514-U
3.02577515-U
3.03577516-U
4.62577517-U
4.63577518-U
3 µM Discovery HS F5 Columns
2.1 3.3567501-U
3.0 3.3567505-U
4.6 3.3567509-U



How to Contact Supelco

In USA:

Phone: 800-247-6628 or 814-359-3441
fax: 800-325-5052
email: techservice@sial.com

Outside the USA: click here

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