HPLC

Discovery® HPLC Columns for Fast LC-MS Analysis

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Ordering Information
 
LC-MS Columns
 
Discovery® HPLC Columns for Fast LC-MS Analysis
New: HS F5 available in short column format.
 
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, <10ppm metals   Spherical, high purity, <10ppm metals
Particle Size:   5µm   5µm
Pore Size:   180Å   180Å
Surface Area:   200m2/g   200m2/g
%C:   12%   7.5%
Coverage:   3µmoles/m2   3.4µmoles/m2
    C18 structure   C8 structure
 
  Properties     CYANO     HS F5
Bonded Phase:   Cyanopropylsilane, endcapped   Pentafluorophenylpropyl, endcapped
Silica:   Spherical, high purity, <10ppm metals   Spherical, high purity, <10ppm metals
Particle Size:   5µm   3.5µm
Pore Size:   180Å   120Å
Surface Area:   200m2/g   300m2/g
%C:   4.5%   12%
Coverage:   3.5µmoles/m2   4µmoles/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, 15cm x 2.1mm ID, 5µm particles
Cat. No.:   50495521
Column 2:   Discovery C18, 2cm x 2.1mm ID, 5µm particles
Cat. No.:   577507-U
Mobile Phase:   60:40, 33mM HCO2H:CH3CN
Flow Rate:   0.2mL/min
Temp.:   35ºC
Det.:   ESI(+), full scan
Inj.1:   6µL
Inj.2:   0.8µL
Sample:   10mg/L in 50% CH3OH
     
 

 
 
Figure 2. Application of Short, Fast LC-MS Columns for Analysis of Veterinary Antibiotics on Discovery C18 and C8 Columns
 
     
Column 1:   Discovery C18, 2cm x 2.1mm ID, 5µm particles
Cat. No.:   577507-U
Column 2:   Discovery C8, 2cm x 2.1mm ID, 5µm particles
Cat. No.:   577501-U
Mobile Phase:   A=50mM HOAc/NH4OH, (pH 6.0), B=water, C=CH3OH
Flow Rate:   0.2mL/min
Temp.:   65ºC
Inj.:   1µL
Det.:   ESI(-)
Sample:    
1.   Lasalocid A (1µg/mL)
2. 3.   Salinomycin (10µg/mL)
4.   Narasin (5µg/mL)
 
Discovery C18, A:B:C = 20:5:75 Discovery C8, A:B:C = 20:10:70
   

Excellent Peak Shape
 
Figure 3. Application of Short, Fast LC-MS Columns for Analysis of Erythromycin c and Clarithromycin on Discovery C18, C8, and Cyano Columns
 
     
Column 1:   Discovery C18, 2cm x 2.1mm ID, 5µm particles
Cat. No.:   577507-U
Column 2:   Discovery C8, 2cm x 2.1mm ID, 5µm particles
Cat. No.:   577501-U
Column 3:   Discovery Cyano, 3cm x 2.1mm ID, 5µm particles
Cat. No.:   577514-U
Mobile Phase:   A=50mM HCO2H, B=water, C=90:10 CH3OH:CH3CN
Flow Rate:   0.2mL/min
Temp.:   35ºC
Inj.:   1µL
Det.:   ESI(+)
Sample:    
1.   Erythromycin c (1µg/mL)
2.   Clarithromycin (1µg/mL)
 
Discovery C18
A:B:C = 20:35:45
Discovery C8
A:B:C = 20:35:45
Discovery Cyano
A:B:C = 20:40:40
     

 
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.
 
Table 1. Rapid Gradient Turnaround Testing of Discovery LC-MS HPLC Columns
Phase Efficiency
(N/column)
USP Tailing
Factor
Retention Time
(min.)
Pressure
(bar)
Discovery C18 (3cm x 3.0mm ID)
Before stability test 2792 1.04 1.989 39
After stability test 2899 1.09 1.985 41
 
Discovery C8 (2cm x 2.1mm ID)
Before stability test 1451 1.05 1.767 26
After stability test 1507 1.09 1.761 25
 
Discovery Cyano (3cm x 4.6mm ID)
Before stability test 3329 1.01 1.368 44
After stability test 3448 1.01 1.362 40

 

Table 2. Test Conditions
Discovery C18 (3cm x 3.0mm ID)
Flow:   0.8mL/min
Gradient:   10-90% methanol over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.:   30°C
Wavelength:   254nm
Pressure:   76bar for first cycle
75bar after last cycle
No. of Cycles:   1,135 three-minute
Total Volume:   2724mL
Void Volume:   0.447
 
Discovery C8 (2cm x 2.1mm ID)
Flow:   0.5mL/min
Gradient:   10-90% methanol over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.:   30°C
Wavelength:   254nm
Pressure:   40bar after first cycle
38bar after last cycle
No. of Cycles:   1,089 three-minute
Total Volume:   1634mL
Void Volume:   0.393
 
Discovery Cyano (3cm x 4.6mm ID)
Flow:   0.2mL/min
Gradient:   10-50% acetnonitrile over 2 minutes back to 10% for 1 minute equilibrium for each 3 minute cycle
Temp.:   30°C
Wavelength:   254nm
Pressure:   72bar after first cycle
70bar after last cycle
No. of Cycles:   937 three-minute
Total Volume:   5622mL
Void Volume:   0.434

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

 
Figure 4. LC-MS Bleed Study
 
Blank Run (no LC column)
(19-21 min.)
Discovery C18 column
(19-21 min.)


Ordering Information
Phase ID (mm) Length (cm) Cat. No.
5µm Discovery C8 Columns
  2.1 2 577501-U
  2.1 3 577502-U
  3.0 2 577503-U
  3.0 3 577504-U
  4.6 2 577505-U
  4.6 3 577506-U
5µm Discovery C18 Columns
  2.1 2 577507-U
  2.1 3 577508-U
  3.0 2 577509-U
  3.0 3 577510-U
  4.6 2 577511-U
  4.6 3 577512-U
5µm Discovery Cyano Columns
  2.1 2 577513-U
  2.1 3 577514-U
  3.0 2 577515-U
  3.0 3 577516-U
  4.6 2 577517-U
  4.6 3 577518-U
3µm Discovery HS F5 Columns
  2.1    3.3 567501-U
  3.0    3.3 567505-U
  4.6    3.3 567509-U

 
Related Information
 
Discovery C18 and C8 Columns - entire family
Discovery Cyano Columns - entire family
Discovery HS F5 Columns - entire family
   
view the pdf document Application Report 46 - Drugs of Abuse on Discovery HS F5 (T003046) (421 KB)
view the pdf document Application Report 134 - Separation of Dopamine and Related Compounds using Discovery HS F5 with Mass Spectrometric Detection (T004134) (17 KB)
view the pdf document Application Report 136 - Resolution of Bilirubin and Biliverdin on Discovery C18 (T004136) [Note: With MS compatible mobile phase.] (16 KB)
   
view the pdf document 2004 Poster Presentation - Advantages of Polar, Reversed-Phase HPLC Columns for the Analysis of Drug Metabolites (T404045) (174 KB)
view the pdf document 2004 Poster Presentation - Complementary Selectivity of Peptides on Aliphatic versus Fluorinated HPLC Phases (T404070) (158 KB)

 
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