Analytical / Chromatography > HPLC > Columns > Discovery^® > Discovery LC MS

HPLC

Discovery LC MS

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.

Read the applications report "Sensitive LC-Analysis of Basic Compounds Using Discovery HS F5" found in TheReporter Vol. 22.5.

Discovery LC-MS Columns Deliver...

High speed separations
Excellent peak shape
Demonstrated ruggedness, stability and reproducibility
Excellent LC-MS bleed characteristics

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


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

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 CHROMASOLV from Riedel-de Haën, 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

	Product Profile - Discovery LC-MS Columns - (T404073) (88 KB)
	Application Report 46 - Drugs of Abuse on Discovery HS F5 (T003046) (421 KB)
	Application Report 134 - Separation of Dopamine and Related Compounds using Discovery HS F5 with Mass Spectrometric Detection (T004134) (17 KB)
	Application Report 136 - Resolution of Bilirubin and Biliverdin on Discovery C18 (T004136) [Note: With MS compatible mobile phase.] (16 KB)
	Application Note 179 - LC-MS CHROMASOLV^® Solvents (T304179) (81 KB)
	2004 Poster Presentation - Advantages of Polar, Reversed-Phase HPLC Columns for the Analysis of Drug Metabolites (T404045) (174 KB)
	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

TRADEMARKS

Discovery and CHROMASOLV are trademarks of Sigma-Aldrich Co.

your favorite gene search

Service & Support

Ordering

Corporate

©2011 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA, SAFC, SIGMA-ALDRICH, ALDRICH, and SUPELCO are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. FLUKA is a trademark of Sigma-Aldrich GmbH, registered in the US and other countries. Sigma brand products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Site Use Terms | Privacy