Back to Table of Contents

Innovators in life and laboratory sciences, Thermo Electron Corporation provides Thermo Electron Corporation
has direct subsidiary offices in BETASIL™ HPLC Columns
North America, Europe and Japan.
To complement these
advanced analytical technologies, scientific instrumentation, laboratory informatics direct subsidiaries, we maintain
a network of representative

solutions, and laboratory consumables to help scientists and clinicians to discover

organizations throughout the
world. Use this reference list or
Technical Guide
visit our web site to locate the
representative nearest you.

new drugs, improve manufacturing processes, and diagnose illness and disease.

France
Unparalleled in our capabilities, we can help you every step of the way – from sample Tel. (33) 1 60 92 48 00

Germany
preparation and sample analysis through interpretation of results. Tel. (49) 6103 4080

United Kingdom Highly Efficient, Retentive and Selective HPLC Columns

Tel. (44) 1928 581000

United States
Tel. (01) 800 437 2999

For more information on our products and services,

please visit our website at: www.thermo.com/chromatography

Technical information contained in this publication is for
reference purposes only and is subject to change without
notice. Every effort has been made to supply complete and
accurate information; however, Thermo Electron assumes no
responsibility and will not be liable for any errors, omissions,
damage, or loss that might result from any use of the
information contained therein (even if this information is
properly followed and problems still arise).
Reference to specifications supersede all previous information
and are subject to change without notice.
ADVANCE, BetaBasic, BetaMax, BETASIL, BioBasic, DASH,
DELTABOND, Duet, Fluophase, Hyperbond, Hypercarb, KAPPA,
HOT POCKET, HyperGEL, HyperREZ, HyperSEP, Hypersil,
HyPURITY, HyPURITY AQUASTAR, Javelin, KEYSTONE, MultiSEP,
PIONEER, PRISM, Retain, SLIPFREE, UNIGUARD, UNIPHASE, and
Verify are trademarks of Thermo Electron Corporation and its
subsidiaries.
AquaSilTM Siliconizing Fluid for treating glass surfaces is sold by
Pierce Chemical Co., Rockford, IL.
All other trademarks are the property of their respective owners.

www.thermo.com/hypersil-keystone
©2003 Thermo Electron, Printed in USA 05/03.
 Back to Table of Contents

BETASIL Columns
Technical Guide
Common Analgesic Water-Soluble Vitamins Pesticides
BETASIL™ Columns Sample:
Sample: 1. Dimethoate
Sample: 2. Methyl Parathion
1. Pyridoxine
1. Acetaminophen 3. Malathion
2. Thiamine
Introduction 2. Caffeine
3. Niacinamide 4. Fenitrothion
3. Acetylsalicylic Acid 5. Ethyl Parathion (DNTP)
BETASIL columns from Thermo 4. Cyanocobalamin
4. Benzoic Acid 6. Diazinon
2 5. Riboflavin
Electron are based on a highly pure, Specifications 7. EPN
4
100Å silica designed for small Phase Particle size Carbon Load Pore Size Endcapping Silica type
3
molecule analysis. All reversed 1 1
BETASIL C18 3, 5 and 10µm 20% 100Å Yes High purity,
base deactivated
phases are end-capped with high
BETASIL C8 3, 5 and 10µm 12% 100Å Yes High purity,
phase loading, providing excep- base deactivated
1
tional chromatography of acids, BETASIL C6 3 and 5µm 11% 100Å Yes High purity, 4
7
bases and neutral compounds. The base deactivated 3
4 2
5
BETASIL C1 3 and 5µm 4% 100Å Yes High purity, 2 5
dense bonding means that mobile base deactivated
3 6
phases require more organic BETASIL Phenyl 5µm 11% 100Å Yes High purity,
solvent, making these columns a base deactivated
popular choice for LC/MS applica- BETASIL Phenyl/Hexyl 3 and 5µm 11% 100Å Yes High purity,
base deactivated
tions. BETASIL Cyano 5µm 7% 100Å Yes High purity,
base deactivated
100Å high purity silica 701-068
BETASIL Diol 100 5µm 6% 100Å Yes High purity, vit10050 beta0031
High surface area and phase loading base deactivated
0 2 MIN 0 5 10 MIN 0 5 10 15 20 25 MIN
BETASIL Silica 100 5µm N/A 100Å N/A High purity,
Dense bonding results in excellent base deactivated BETASIL® C18 PIONEERTM, 5µm, 50x3mm BETASIL C18, 5µm, 150x4.6mm BETASIL C18, 5µm, 150x4.6mm
reproducibility and stability Eluent: A: 0.05M Ammonium Acetate, pH 4 Gradient: A: 0.05M K2HPO4, pH 6.5 Gradient: 50% ACN / 50% H2O, step to
B: 70% ACN% / 30% MeOH B: ACN 60% ACN / 40% H2O at 12 min.
85:15 A:B 2% B to 20% B in 15 min Flow: 1.0 mL/min
Excellent performance with acids, bases Flow: 1.0 mL/min
Flow: 2.5 mL/min Detector: UV @ 235
and neutral compounds Detector: UV@ 254 Detector: UV @ 265

BETASIL Chromatographic Figure 1. Increased Hydrophobic Retention of Neutral Molecules

Steroids by Normal Phase
Characterization
Sample: Sample:
The BETASIL C18 phase is a high 7. 11α-Ketoprogesterone 1 3
1. Uracil 5. Butyl Benzene 1. Progesterone
2. Benzene 6. Pentyl Benzene 2. 20α-Hydroxy-4-pregnene-3-one 8. Corticosterone
carbon load packing that offers up 3. Ethyl Benzene 7. Hexyl Benzene 3. 17α-Hydroxyprogesterone 9. Cortisone Acetate
to twice the retention of other 4. Propyl Benzene 8. Heptyl Benzene 1 4. Deoxycorticosterone 10. Hydrocortisone
5. 11α-Hydroxyprogesterone Acetate 11. Cortisone 6
C18 packings with lower percent BETASIL C18 BetaBasic 18 Hypersil BDS C18 6. 11α-Hydroxyprogesterone
3
carbon loading. Figure 1 shows 20%C 13%C 11%C 4,7 6
the increased retention of the 1 3 1 1 1 2 5
BETASIL C18 column when 2 4,7
compared to other Thermo 2
3 5 8
Electron columns. The increased 4 4,7 6
2 9 10
retention associated with the 2 2 5 8 10

BETASIL column is directly 5 3 9 9

8
attributed to the increased surface 4
11
6 5 10 11
area of theparticles, and higher 3

carbon loading than both 7 4 6

5 11
Hypersil™ BDS C18 and 6 7
8
7
BetaBasic™ 18 columns. 8

Where longer retention is 701-093 715-173 046-042 725-001 726-001 728-001

required or there is a requirement 0 5 10 15 20 MIN 0 5 10 15 MIN 0 5 10 15 MIN 0 5 10 MIN 0 5 10 MIN 0 5 MIN
to use a higher percentage of
BETASIL Diol 60, 5µm, 150x4.6mm BETASIL Diol 100, 5µm, 150x4.6mm BETASIL Diol 300, 5µm, 150x4.6mm
organic solvent in themobile Columns: 5µm, 150x4.6mm
Eluent: 85% Isooctane / 14.7% EtOH / 0.3% H2O Eluent: 85% Isooctane / 14.7% EtOH Eluent: 85% Isooctane / 14.7%
Eluent: 75% ACN / 25% H2O
phase, e.g. to increase LC/MS Flow: 1.25 mL/min
Flow: 1.2 mL/min / 0.3% H2O EtOH / 0.3% H2O
Detector: UV @ 254 Flow: 1.2 mL/min Flow: 1.2 mL/min
sensitivity, the BETASIL C18 Detector: UV @ 254
Detector: UV @ 254 Detector: UV @ 254
packing offers an excellent choice.

2 7
 Back to Table of Contents
Stability at Low pH Figure 2. BETASIL Phase Stability At Low pH

BETASIL Columns
Technical Guide
Carbamate/Urea Procainamides All surface bonded chemistries
Pesticides & Herbicides 1 0.1% TFA gradients at 50°C, pH 1.8
Sample:
belonging to the BETASIL family are
Sample: 1. Uracil highly stable under chemical attack.
2. Procainamide
1. Tebuthiuron
3. N-Acetyl Procainamide
Hydrolysis of the silane ligand can
2. Carbofuran
3. Carbaryl 4. Caffeine occur under acidic conditions,
5. Propionyl Procainamide
4. Diuron resulting in a loss of retention and/
5. Barban 2
4
or selectivity. The effect becomes
3 more severe as ligand chain length
is reduced, as is observed for the
BETASIL C1 phase. Wettable
1 stationary phases such as the
BETASIL Cyano packing can also
show slightly less resistance to
4 chemical attack, and consequently
3 are recommended only for use
5
between pH 2 and 8.
5
In general, BETASIL packings show
2 superior stability to many other
packings. Figure 2 shows BETASIL reversible loss of retention when retention loss is that the hydropho-
H730-005 708-002
phases subjected to 25,000 exposed to highly aqueous mobile bic alkyl chains of the stationary
0 2 4 6 8 10 MIN 0 4 8 MIN column volumes of aggressive phase. The rate and degree of phase are not wettable and appear
BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm BETASIL C6, 5µm, 150x4.6mm mobile phase at pH 1.8 and 50°C. retention loss can vary greatly to “fold” down on the silica surface
Gradient: A: H2O Eluent: 10% ACN / 90% 0.05M KH2PO4,
The BETASIL C18 phase exhibits among different columns. A to avoid a highly aqueous, hydro-
B: ACN pH 3.5 with H3PO4
Time %B Flow: 1.25 mL/min virtually no change in retention for common explanation for this philic mobile phase (Figure 4).
0 15 Detector: UV@ 254
7 85
the analysis of a sensitive drug
Figure 3. Base Deactivation of BETASIL C6 Figure 4. Phase Collapse
Flow: 1.5 mL/min mixture.
Detector: UV @ 254 Sample:
Temp.: 25°C 1. Uracil
Base Deactivated for Better Peak 2. Procainamide
Shape with Basic Compounds 3. N-Acetyl Procainamide
4. Caffeine
Cyclosporin Assay All BETASIL packings are base 5. N-Propionyl Procainamide
1 Real Sample: deactivated and show excellent
500 ng / mL 383 ng / mL peak shape for basic compounds. 2
standard Sample:
Figure 3 shows the analysis of a
1. Cyclosporin 1
2. Internal Standard procainamide test mixture, a
2 1 4 5
mixture that would typically have
2 3
given rise to broad tailing peaks for
the more basic compounds on
traditional C18 silicas prepared on a
Type A silica. In terms of silanol
activity, the BETASIL packing is
similar to other Type B silica-based
phases such as BetaBasic and
BetaMax™ Neutral phases.
703-002
c1-std c1-pat Phase Collapse Phenomenon
Mobile phases that contain a high 0 5 MIN
0 5 MIN 0 5 MIN
proportion of water often are
BETASIL C6, 5µm, 150x4.6mm
BETASIL C1, 5µm, 150x4.6mm employed to retain highly polar Eluent: 10% ACN / 90% 0.05M Phosphate
Eluent: 35% ACN / 65% 0.05M KH2PO4, pH 7
Flow: 1.25 mL/min
compounds when using RP-HPLC. buffer, pH 3.5 with H3PO4
Flow: 1.25 mL/min
Detector: UV @ 254 Many C18 columns will show a Detector: UV @ 254

6 3
 Back to Table of Contents

BETASIL Columns
Technical Guide
Regeneration After Phase Collapse
In this folded or collapsed state,
the alkyl chains are much less able
to interact with solutes, resulting
in a loss of retention. We use the
term “chain folding” to refer to
this reversible loss of retention
induced by highly aqueous mobile
phases, although the exact
mechanism of reversible retention
Figure 6. Trace Organics in
classical reversed phase reten- Mobile Phase Retard Chain Folding Figure 7. Polyphenols
The phenomena of chain folding
can be almost completely over- tion, while the phenyl ring Sample:
provides special selectivity for 1. Tartaric acid
come by including trace amounts 2. Malonic acid 2 Sample:
of organic solvent in the mobile polar groups (Fig. 7). 3. Fumaric acid 1. Pyrogallol
4. Succinic acid 2. Hydroquinone
phase. Figure 6 shows how just 4
3. Resorcinol
0.3% n-propanol has been used to BETASIL columns also offer 4. Phenol
retard the loss in retention choices for normal phase chroma- 2 3 Initial
associated with chain collapse. tography, including silica and diol
phases in 4 different pore sizes 1

loss is not proven. Choices in Stationary Phase from 60Å to 300Å, and cyano.
The BETASIL phase is a high Chemistries Diol phases provide higher polar 3
bet18016
carbon load, high density C18 and BETASIL columns are available in a selectivity than cyano with less
consequently shows the charac- wide range of reversed phase water sensitivity than bare silica. 4
teristics associated with chain packings, including C18, C8, C6, 1
folding, i.e. loss in retention. Phenyl, Cyano, and Phenyl/Hexyl. 2 4
However, the situation is com- The BETASIL Phenyl/Hexyl phase 3 After 8 hours
pletely reversible and the column provides a combination of straight- 1 H730-007
can be regenerated simply by chain C6 groups and phenyl
0 5 10 15 20 25 30 MIN
flushing with 40% ACN / 60% H2O groups, resulting in a mixed-mode
for 10 minutes (Figure 5). separation. The C6 chain exhibits bet18020 BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm
Eluent: 5mM Acetic Acid
Flow: 1.0 mL/min
0 2 4 6 MIN Detector: UV @ 280
Temp.: 25°C
BETASIL C18, 150x4.6mm
Figure 5. High Carbon Load C18 in 100% Aqueous Mobile Phase Eluent: 0.05M KH2PO4 + 0.03M H3PO 4 + 0.3%
n-propanol
Flow: 1.0 mL/min
Detector: UV @ 210
Sample: 4
Organic Acids Benzodiazepines
4 1,2,3,4
1. Tartaric acid 2
2. Malonic acid Sample: 4
3. Fumaric acid 1. Nitrazepam
4. Succinic acid Sample:
1. Ascorbic Acid 2. Oxazepam
2. Acetaminophen 3. Temazepam
3 4. Nordiazepam 3
3 3. Benzoic Acid 5
4. Salicylic Acid 5. Diazepam 2
6. Medazepam

2 1

1 6
1
2
1

bet18001 bet18004 bet18005

3 4

0 2 4 6 8 MIN 0 2 4 6 8 MIN 0 2 4 6 8 MIN H730-006 H730-008

0 2 4 6 8 10 12 14 16 MIN 0 2 4 6 MIN
Partially Folded Completely Folded Regenerated (40% ACN / BETASIL® Phenyl/Hexyl, 5µm, 150x4.6mm BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm
after >2 hours use 60% H2O for 10 minutes) Eluent: 80% 20mM KH2PO4, pH 2 / 20% MeOH Gradient: A: H2O
Flow: 1.0 mL/min B: MeOH
BETASIL C18, 150x4.6mm Detector: UV @ 254 Time %B
Eluent: 0.05M KH2PO4 + 0.03M H3PO4 Temp.: 25°C 0 15
Flow: 1.0 mL/min 7 85
Detector: UV @ 210 Flow: 1.0 mL/min
Detector: UV @ 235
Temp.: 25°C

4 5
 Back to Table of Contents

BETASIL Columns
Technical Guide
Regeneration After Phase Collapse
In this folded or collapsed state,
the alkyl chains are much less able
to interact with solutes, resulting
in a loss of retention. We use the
term “chain folding” to refer to
this reversible loss of retention
induced by highly aqueous mobile
phases, although the exact
mechanism of reversible retention
Figure 6. Trace Organics in
classical reversed phase reten- Mobile Phase Retard Chain Folding Figure 7. Polyphenols
The phenomena of chain folding
can be almost completely over- tion, while the phenyl ring Sample:
provides special selectivity for 1. Tartaric acid
come by including trace amounts 2. Malonic acid 2 Sample:
of organic solvent in the mobile polar groups (Fig. 7). 3. Fumaric acid 1. Pyrogallol
4. Succinic acid 2. Hydroquinone
phase. Figure 6 shows how just 4
3. Resorcinol
0.3% n-propanol has been used to BETASIL columns also offer 4. Phenol
retard the loss in retention choices for normal phase chroma- 2 3 Initial
associated with chain collapse. tography, including silica and diol
phases in 4 different pore sizes 1

loss is not proven. Choices in Stationary Phase from 60Å to 300Å, and cyano.
The BETASIL phase is a high Chemistries Diol phases provide higher polar 3
bet18016
carbon load, high density C18 and BETASIL columns are available in a selectivity than cyano with less
consequently shows the charac- wide range of reversed phase water sensitivity than bare silica. 4
teristics associated with chain packings, including C18, C8, C6, 1
folding, i.e. loss in retention. Phenyl, Cyano, and Phenyl/Hexyl. 2 4
However, the situation is com- The BETASIL Phenyl/Hexyl phase 3 After 8 hours
pletely reversible and the column provides a combination of straight- 1 H730-007
can be regenerated simply by chain C6 groups and phenyl
0 5 10 15 20 25 30 MIN
flushing with 40% ACN / 60% H2O groups, resulting in a mixed-mode
for 10 minutes (Figure 5). separation. The C6 chain exhibits bet18020 BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm
Eluent: 5mM Acetic Acid
Flow: 1.0 mL/min
0 2 4 6 MIN Detector: UV @ 280
Temp.: 25°C
BETASIL C18, 150x4.6mm
Figure 5. High Carbon Load C18 in 100% Aqueous Mobile Phase Eluent: 0.05M KH2PO4 + 0.03M H3PO 4 + 0.3%
n-propanol
Flow: 1.0 mL/min
Detector: UV @ 210
Sample: 4
Organic Acids Benzodiazepines
4 1,2,3,4
1. Tartaric acid 2
2. Malonic acid Sample: 4
3. Fumaric acid 1. Nitrazepam
4. Succinic acid Sample:
1. Ascorbic Acid 2. Oxazepam
2. Acetaminophen 3. Temazepam
3 4. Nordiazepam 3
3 3. Benzoic Acid 5
4. Salicylic Acid 5. Diazepam 2
6. Medazepam

2 1

1 6
1
2
1

bet18001 bet18004 bet18005

3 4

0 2 4 6 8 MIN 0 2 4 6 8 MIN 0 2 4 6 8 MIN H730-006 H730-008

0 2 4 6 8 10 12 14 16 MIN 0 2 4 6 MIN
Partially Folded Completely Folded Regenerated (40% ACN / BETASIL® Phenyl/Hexyl, 5µm, 150x4.6mm BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm
after >2 hours use 60% H2O for 10 minutes) Eluent: 80% 20mM KH2PO4, pH 2 / 20% MeOH Gradient: A: H2O
Flow: 1.0 mL/min B: MeOH
BETASIL C18, 150x4.6mm Detector: UV @ 254 Time %B
Eluent: 0.05M KH2PO4 + 0.03M H3PO4 Temp.: 25°C 0 15
Flow: 1.0 mL/min 7 85
Detector: UV @ 210 Flow: 1.0 mL/min
Detector: UV @ 235
Temp.: 25°C

4 5
 Back to Table of Contents
Stability at Low pH Figure 2. BETASIL Phase Stability At Low pH

BETASIL Columns
Technical Guide
Carbamate/Urea Procainamides All surface bonded chemistries
Pesticides & Herbicides 1 0.1% TFA gradients at 50°C, pH 1.8
Sample:
belonging to the BETASIL family are
Sample: 1. Uracil highly stable under chemical attack.
2. Procainamide
1. Tebuthiuron
3. N-Acetyl Procainamide
Hydrolysis of the silane ligand can
2. Carbofuran
3. Carbaryl 4. Caffeine occur under acidic conditions,
5. Propionyl Procainamide
4. Diuron resulting in a loss of retention and/
5. Barban 2
4
or selectivity. The effect becomes
3 more severe as ligand chain length
is reduced, as is observed for the
BETASIL C1 phase. Wettable
1 stationary phases such as the
BETASIL Cyano packing can also
show slightly less resistance to
4 chemical attack, and consequently
3 are recommended only for use
5
between pH 2 and 8.
5
In general, BETASIL packings show
2 superior stability to many other
packings. Figure 2 shows BETASIL reversible loss of retention when retention loss is that the hydropho-
H730-005 708-002
phases subjected to 25,000 exposed to highly aqueous mobile bic alkyl chains of the stationary
0 2 4 6 8 10 MIN 0 4 8 MIN column volumes of aggressive phase. The rate and degree of phase are not wettable and appear
BETASIL Phenyl/Hexyl, 5µm, 150x4.6mm BETASIL C6, 5µm, 150x4.6mm mobile phase at pH 1.8 and 50°C. retention loss can vary greatly to “fold” down on the silica surface
Gradient: A: H2O Eluent: 10% ACN / 90% 0.05M KH2PO4,
The BETASIL C18 phase exhibits among different columns. A to avoid a highly aqueous, hydro-
B: ACN pH 3.5 with H3PO4
Time %B Flow: 1.25 mL/min virtually no change in retention for common explanation for this philic mobile phase (Figure 4).
0 15 Detector: UV@ 254
7 85
the analysis of a sensitive drug
Figure 3. Base Deactivation of BETASIL C6 Figure 4. Phase Collapse
Flow: 1.5 mL/min mixture.
Detector: UV @ 254 Sample:
Temp.: 25°C 1. Uracil
Base Deactivated for Better Peak 2. Procainamide
Shape with Basic Compounds 3. N-Acetyl Procainamide
4. Caffeine
Cyclosporin Assay All BETASIL packings are base 5. N-Propionyl Procainamide
1 Real Sample: deactivated and show excellent
500 ng / mL 383 ng / mL peak shape for basic compounds. 2
standard Sample:
Figure 3 shows the analysis of a
1. Cyclosporin 1
2. Internal Standard procainamide test mixture, a
2 1 4 5
mixture that would typically have
2 3
given rise to broad tailing peaks for
the more basic compounds on
traditional C18 silicas prepared on a
Type A silica. In terms of silanol
activity, the BETASIL packing is
similar to other Type B silica-based
phases such as BetaBasic and
BetaMax™ Neutral phases.
703-002
c1-std c1-pat Phase Collapse Phenomenon
Mobile phases that contain a high 0 5 MIN
0 5 MIN 0 5 MIN
proportion of water often are
BETASIL C6, 5µm, 150x4.6mm
BETASIL C1, 5µm, 150x4.6mm employed to retain highly polar Eluent: 10% ACN / 90% 0.05M Phosphate
Eluent: 35% ACN / 65% 0.05M KH2PO4, pH 7
Flow: 1.25 mL/min
compounds when using RP-HPLC. buffer, pH 3.5 with H3PO4
Flow: 1.25 mL/min
Detector: UV @ 254 Many C18 columns will show a Detector: UV @ 254

6 3
 Back to Table of Contents

BETASIL Columns
Technical Guide
Common Analgesic Water-Soluble Vitamins Pesticides
BETASIL™ Columns Sample:
Sample: 1. Dimethoate
Sample: 2. Methyl Parathion
1. Pyridoxine
1. Acetaminophen 3. Malathion
2. Thiamine
Introduction 2. Caffeine
3. Niacinamide 4. Fenitrothion
3. Acetylsalicylic Acid 5. Ethyl Parathion (DNTP)
BETASIL columns from Thermo 4. Cyanocobalamin
4. Benzoic Acid 6. Diazinon
2 5. Riboflavin
Electron are based on a highly pure, Specifications 7. EPN
4
100Å silica designed for small Phase Particle size Carbon Load Pore Size Endcapping Silica type
3
molecule analysis. All reversed 1 1
BETASIL C18 3, 5 and 10µm 20% 100Å Yes High purity,
base deactivated
phases are end-capped with high
BETASIL C8 3, 5 and 10µm 12% 100Å Yes High purity,
phase loading, providing excep- base deactivated
1
tional chromatography of acids, BETASIL C6 3 and 5µm 11% 100Å Yes High purity, 4
7
bases and neutral compounds. The base deactivated 3
4 2
5
BETASIL C1 3 and 5µm 4% 100Å Yes High purity, 2 5
dense bonding means that mobile base deactivated
3 6
phases require more organic BETASIL Phenyl 5µm 11% 100Å Yes High purity,
solvent, making these columns a base deactivated
popular choice for LC/MS applica- BETASIL Phenyl/Hexyl 3 and 5µm 11% 100Å Yes High purity,
base deactivated
tions. BETASIL Cyano 5µm 7% 100Å Yes High purity,
base deactivated
100Å high purity silica 701-068
BETASIL Diol 100 5µm 6% 100Å Yes High purity, vit10050 beta0031
High surface area and phase loading base deactivated
0 2 MIN 0 5 10 MIN 0 5 10 15 20 25 MIN
BETASIL Silica 100 5µm N/A 100Å N/A High purity,
Dense bonding results in excellent base deactivated BETASIL® C18 PIONEERTM, 5µm, 50x3mm BETASIL C18, 5µm, 150x4.6mm BETASIL C18, 5µm, 150x4.6mm
reproducibility and stability Eluent: A: 0.05M Ammonium Acetate, pH 4 Gradient: A: 0.05M K2HPO4, pH 6.5 Gradient: 50% ACN / 50% H2O, step to
B: 70% ACN% / 30% MeOH B: ACN 60% ACN / 40% H2O at 12 min.
85:15 A:B 2% B to 20% B in 15 min Flow: 1.0 mL/min
Excellent performance with acids, bases Flow: 1.0 mL/min
Flow: 2.5 mL/min Detector: UV @ 235
and neutral compounds Detector: UV@ 254 Detector: UV @ 265

BETASIL Chromatographic Figure 1. Increased Hydrophobic Retention of Neutral Molecules

Steroids by Normal Phase
Characterization
Sample: Sample:
The BETASIL C18 phase is a high 7. 11α-Ketoprogesterone 1 3
1. Uracil 5. Butyl Benzene 1. Progesterone
2. Benzene 6. Pentyl Benzene 2. 20α-Hydroxy-4-pregnene-3-one 8. Corticosterone
carbon load packing that offers up 3. Ethyl Benzene 7. Hexyl Benzene 3. 17α-Hydroxyprogesterone 9. Cortisone Acetate
to twice the retention of other 4. Propyl Benzene 8. Heptyl Benzene 1 4. Deoxycorticosterone 10. Hydrocortisone
5. 11α-Hydroxyprogesterone Acetate 11. Cortisone 6
C18 packings with lower percent BETASIL C18 BetaBasic 18 Hypersil BDS C18 6. 11α-Hydroxyprogesterone
3
carbon loading. Figure 1 shows 20%C 13%C 11%C 4,7 6
the increased retention of the 1 3 1 1 1 2 5
BETASIL C18 column when 2 4,7
compared to other Thermo 2
3 5 8
Electron columns. The increased 4 4,7 6
2 9 10
retention associated with the 2 2 5 8 10

BETASIL column is directly 5 3 9 9

8
attributed to the increased surface 4
11
6 5 10 11
area of theparticles, and higher 3

carbon loading than both 7 4 6

5 11
Hypersil™ BDS C18 and 6 7
8
7
BetaBasic™ 18 columns. 8

Where longer retention is 701-093 715-173 046-042 725-001 726-001 728-001

required or there is a requirement 0 5 10 15 20 MIN 0 5 10 15 MIN 0 5 10 15 MIN 0 5 10 MIN 0 5 10 MIN 0 5 MIN
to use a higher percentage of
BETASIL Diol 60, 5µm, 150x4.6mm BETASIL Diol 100, 5µm, 150x4.6mm BETASIL Diol 300, 5µm, 150x4.6mm
organic solvent in themobile Columns: 5µm, 150x4.6mm
Eluent: 85% Isooctane / 14.7% EtOH / 0.3% H2O Eluent: 85% Isooctane / 14.7% EtOH Eluent: 85% Isooctane / 14.7%
Eluent: 75% ACN / 25% H2O
phase, e.g. to increase LC/MS Flow: 1.25 mL/min
Flow: 1.2 mL/min / 0.3% H2O EtOH / 0.3% H2O
Detector: UV @ 254 Flow: 1.2 mL/min Flow: 1.2 mL/min
sensitivity, the BETASIL C18 Detector: UV @ 254
Detector: UV @ 254 Detector: UV @ 254
packing offers an excellent choice.

2 7
 Back to Table of Contents

Innovators in life and laboratory sciences, Thermo Electron Corporation provides Thermo Electron Corporation
has direct subsidiary offices in BETASIL™ HPLC Columns
North America, Europe and Japan.
To complement these
advanced analytical technologies, scientific instrumentation, laboratory informatics direct subsidiaries, we maintain
a network of representative

solutions, and laboratory consumables to help scientists and clinicians to discover

organizations throughout the
world. Use this reference list or
Technical Guide
visit our web site to locate the
representative nearest you.

new drugs, improve manufacturing processes, and diagnose illness and disease.

France
Unparalleled in our capabilities, we can help you every step of the way – from sample Tel. (33) 1 60 92 48 00

Germany
preparation and sample analysis through interpretation of results. Tel. (49) 6103 4080

United Kingdom Highly Efficient, Retentive and Selective HPLC Columns

Tel. (44) 1928 581000

United States
Tel. (01) 800 437 2999

For more information on our products and services,

please visit our website at: www.thermo.com/chromatography

Technical information contained in this publication is for
reference purposes only and is subject to change without
notice. Every effort has been made to supply complete and
accurate information; however, Thermo Electron assumes no
responsibility and will not be liable for any errors, omissions,
damage, or loss that might result from any use of the
information contained therein (even if this information is
properly followed and problems still arise).
Reference to specifications supersede all previous information
and are subject to change without notice.
ADVANCE, BetaBasic, BetaMax, BETASIL, BioBasic, DASH,
DELTABOND, Duet, Fluophase, Hyperbond, Hypercarb, KAPPA,
HOT POCKET, HyperGEL, HyperREZ, HyperSEP, Hypersil,
HyPURITY, HyPURITY AQUASTAR, Javelin, KEYSTONE, MultiSEP,
PIONEER, PRISM, Retain, SLIPFREE, UNIGUARD, UNIPHASE, and
Verify are trademarks of Thermo Electron Corporation and its
subsidiaries.
AquaSilTM Siliconizing Fluid for treating glass surfaces is sold by
Pierce Chemical Co., Rockford, IL.
All other trademarks are the property of their respective owners.

www.thermo.com/hypersil-keystone
©2003 Thermo Electron, Printed in USA 05/03.