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[Home](https://www.sigmaaldrich.com/US/en)[Small Molecule HPLC](https://www.sigmaaldrich.com/US/en/applications/analytical-chemistry/liquid-chromatography/small-molecule-hplc) Discovery® HPLC Columns for Fast LC-MS Analysis
# 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:
- [High speed separations](https://www.sigmaaldrich.com#hispeed)
- [Excellent peak shape](https://www.sigmaaldrich.com#peakshp)
- [Demonstrated ruggedness, stability and reproducibility](https://www.sigmaaldrich.com#Demonstrated)
- [Excellent LC-MS bleed characteristics](https://www.sigmaaldrich.com#bleedanal)
| | | | | |
|--------------------|---|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 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 |
| | |  | |  |
| | | | | |
| 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 |
| | |  | |  |
[](https://www.sigmaaldrich.com)
### 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 |
__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__ |
| | |
|-------------------|-------------------------------------------------------|
| __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 C8__
__A:B:C = 20:35:45__
__Discovery Cyano__
__A:B:C = 20:40:40__
[](https://www.sigmaaldrich.com)
### 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
FactorRetention 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
__Figure 4.__LC-MS Bleed Study.
__Blank Run (no LC column) (19-21 min.)__
__Discovery C18 column (19-21 min.)__
* * *
## Ordering Information
## 5 µM Discovery C8 Columns
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## 5 µM Discovery C18 Columns
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## 5 µM Discovery Cyano Columns
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
## 3 µM Discovery HS F5 Columns
Sorry, an unexpected error has occurred
Response not successful: Received status code 500
* * *
__Related Information__
[Discovery C18 and C8 HPLC Columns](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/large-molecule-hplc/discovery-c18-c8-hplc-columns)
Discovery C18 and C8 HPLC Columns products offered
[Discovery Cyano HPLC Columns](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/large-molecule-hplc/discovery-cyano-hplc-columns)
Discovery Cyano HPLC Columns products offered by Sigma-Aldrich. Unique retention and selectivity enables better, faster separations.
[HPLC Discovery HS F5](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/small-molecule-hplc/hs-f5)
HPLC Discovery HS F5 and silica-based phases products offered by us.
* * *
## Related Literature
- [Application Report 46 - Drugs of Abuse on Discovery HS F5 (T003046)](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/960/771/11843.pdf)
- [Application Report 134 - Separation of Dopamine and Related Compounds using Discovery HS F5 with Mass Spectrometric Detection (T004134)](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/158/822/11952.pdf)
- [Application Report 136 - Resolution of Bilirubin and Biliverdin on Discovery C18 (T004136) \[Note: With MS compatible mobile phase.\]](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/126/961/11954.pdf)
- [2004 Poster Presentation - Advantages of Polar, Reversed-Phase HPLC Columns for the Analysis of Drug Metabolites (T404045)](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/146/806/11908.pdf)
* * *
## How to Contact Supelco
In USA:
Phone: 800-247-6628 or 814-359-3441
fax: 800-325-5052
email: [techservice@sial.com](mailto:techservice@sial.com)
Outside the USA: [click here](https://www.sigmaaldrich.com/US/en/collections/offices)
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