Professional Documents
Culture Documents
Columnas Symmetry
Columnas Symmetry
Columnas Symmetry
Symmetry Family
of HPLC Columns
Column-to-column and batch-to-batch repro- Increased labor costs due to individual Symmetry brand of HPLC columns have the tightest specifi-
ducibility is not consistent column QC testing cations in the market to provide the most consistent results
Revalidate/redevelop method with each Symmetry columns achieve outstanding year-to-year repro-
new batch of columns ducibility to give long life to established analytical methods
Severe or excessive peak tailing Increased number of experiments to SymmetryShield columns with embedded polar
optimize analytical method technology provide less peak tailing for faster and
easier method development
Column selectivity not resolving Increased number of experiments required SymmetryShield columns are the perfect complement to
critical peaks to optimize method Symmetry columns, providing different selectivity and
enabling column selection to maximize resolution
Difficulty resolving individual compounds in Increased cost and time to isolate Symmetry300 columns demonstrate the highest peak
complex peptide or protein samples compounds capacity of all leading brands
Excellent reproducibility for separating large numbers of
peptides or proteins
Scaling-up analytical method to prepara- Higher costs due to method development SymmetryPrep columns save development time with direct
tive scale requires too many adjustments to time and labor scale-up while maintaining resolution
achieve good isolation and purification Higher cost of equipment, samples and
prep columns
Physical Characteristics
example shown. 3
1
Symmetry Columns
The Benefit of Narrow
C18 Ligand Surface Coverage Specification Ranges
Specification Ranges
3.09 3.31
Symmetry columns have the narrowest Symmetry C18
ligand surface coverage specifications in Column X
2.75 3.75
N K2
( )(
-1
the chromatographer in achieving repro- 1.4 R=
4 ) ( K +1 )
2
ducible results. The surface coverage re- 1.2
Specification Ranges
quirements of Symmetry are the tightest 1 for Symmetry C18 R = resolution
N = number of peaks
specifications in the industry, resulting in 0.8
1.5 2 2.5 3 3.5 4
= alpha value
k = peak retention
minimal shifts in resolution and more repro- C18 Surface Coverage (mol/m2)
ducible batch-to-batch and column-to-
column results. Minimal Changes in Resolution Means More Reproducible Results*
4.14 2.86
Symmetry C18
Specification Y 5.37 0
Long Lifetime
Outstanding Packed Bed Stability10,000 Injections
Columns that last longer save you money. 1 Column: Symmetry C8 3.9 mm x 150 mm, 5 m
3
with Sentry Guard Column
Symmetry columns can last over 10,000 3.9 mm x 20 mm
injections with minimal loss in efficiency, Mobile Phase: Water/methanol/glacial
4 acetic acid 79:20:1
5
minimal increase in backpressure and 2
6
Flow Rate: 1.0 mL/minute
Injection Volume: 10 L
minimal change in retention time. Temperature: 25C
Detector: UV @ 254
Symmetry columns deliver guaranteed Analytes: 1. Sulfanilamide
2. Sulfadiazine
consistent performance optimizing the 3. Sulfathiazole
4. Sulfamerazine
two key factors that control column lifetime 5. Sulfamethazine
6. Succinylsulfathiazole
which are hydrolytic stability of the pack- Injection 10,000
0 2 4 6 8 10
the packed bed. Minutes
The long-term mechanical stability of Symmetry columns is demonstrated in this analysis of sulfa drugs. Over 10,000 injections were run
with minimal loss in column efficiency. The use of the Sentry guard column is critical to achieving these types of column lifetimes.
2
Superior Peak Shape
The Influence of Mobile Phase pH on Peak Shape of Amitriptyline
Regardless of Mobile Phase pH
The performance of any reversed-phase Amitriptyline Tailing Versus Buffer pH
column is determined by two factors.
First, the bonded phase coverage
(hydrophobicity); and second, the activity Conventional C18
Minutes
3
SymmetryShield Columns
Brand DO 1.78
Brand DA 1.9
O Polar Brand I 1.95
Si Group
Brand P 2.39
Brand E 3.6
CH3 CH3
0 1 2 3 4
Reduce interaction with silanols
USP Tailing Factor
Improve water-wettability
a: SymmetryShield RP18
Column: a. SymmetryShield RP18 3.9 x 150 mm
b. Leading Brand C18
The embedded polar group, which is in 1 2 Mobile Phase: 65% methanol; 35% 20 mM
4
close proximity to the silica surface, further KH2PO4/K2HPO4 @ pH 7
Flow Rate: 1.0 mL/minute
reduces the activity of the surface silanols in Detection: UV @ 254 nm
Temperature: 23C
SymmetryShield packing materials. 3
Analytes: 1. Uracil
2. Propranolol
As a result, SymmetryShield columns: USP Tailing Factor = 1.2
3. Butylparaben
4. Dipropylphthalate
AU
5
6 5. Naphthalene
+ Deliver significantly improved peak 6. Amitriptyline
7 7. Acenaphthene
shape and resolution over a broad
pH range
C8 columns Minutes
methods faster and improve the produc- 5 USP Tailing Factor = 1.9
0 5 10 15 20 25 30
Minutes
4
Unique Selectivity
SymmetryShield Columns Deliver Unique Selectivity
Obtaining a good separation is para-
mount. SymmetryShield columns exhibit
less retention for basic compounds due to
lower silanol activity than conventional Leading Brand C8
3.9 x 150 mm column,
C18 and C8 columns. SymmetryShield
straight chain alkyl
columns are an excellent methods devel-
opment tool.
0 2 4 6 8 10
Minutes
5
Symmetry300 Columns
0 5 10 15 20 25
Minutes
6
Sharpest Peaks for Highest
Greater Peak Capacity
Resolution Tryptic Map of Bovine Serum Albumin
The heart of the column is the high purity Column: Symmetry300 C18, 4.6 mm x 50 mm Injection Volume: 20 L
base deactivated silica. The silica used in Mobile Phase: Eluent A: 0.1% TFA in water Temperature: 35 C
Eluent B: 0.1% TFA in acetonitrile Detection: UV @ 214 nm
Symmetry300 columns is synthesized Flow Rate: 1.0 mL/min
Gradient: Time Profile
using ultrapure organic reagents, resulting (min) %A %B
in high purity with very low silanol activity. 0.00 100 0
45 70 30
In addition, when combined with the high
surface coverage of the bonded phase
the result is higher recoveries of proteins
and peptides even when injected at
low concentrations.
Minutes
Symmetry300 columns deliver the
sharpest peaks for biomolecules and
deliver more peak capacity for complex
Unsurpassed Reproducibility
tryptic maps.
Column: Symmetry300 C18 3.9 x 150 mm, 5 m
They are packed to the highest efficiency Cytochrome c Tryptic Map Q.C. Test
Mobile Phase A: 0.05% TFA in water
Waters batch release protocol includes a tryptic map Mobile Phase B: 0.1% TFA in acetonitrile
and reproducibility of any product on the Flow rate: 1.0 mL/min
of cytochrome c which is used to test for reproducibili-
market. This results in improved precision ty of retention time and resolution. In this test, the tryp-
Gradient: Time Profile
(min) %A %B
for impurity and stability assays, more accu- tic map was run on four Symmetry300 columns, 0.00 100 0
each containing a different batch of packing.
rate quantitation, and adaptability to more Injection volume:
45
20 L
72 28
Minutes
Minutes
7
SymmetryPrep Columns
4.00
Symmetry 3.5 m and 5 m columns. An Injection volume: 2,000 L
3.00
HPLC method developed on a Symmetry 2.00
analytical column can be scaled up direct- 20x
1.00
0 4.00
Minutes
8
Impurity Profiling: Resolution
Valerophenone Impurities: Scaling up from
Maintained when Scaling-up
Symmetry 5 m to SymmetryPrep 7 m Columns
SymmetryPrep columns are manufactured
Column: a: Symmetry C18, 3.9 mm x 150 mm, 5 m
under the same rigid standards as b: SymmetryPrep C18, 7.8 mm x 300 mm, 7 m
Mobile Phase:
Symmetry analytical columns which a
Flow Rate:
60% acetonitrile/40% water
a: 0.70 mL/min
allows for direct scale-up while maintaining b: 5.60 mL/min
Sample: 4 mg/mL valerophenone
resolution as shown. This can be a Injection Volume: a: 100 L
b: 800 L
significant benefit in saving both time and Detection: UV @ 340 nm
labor costs when isolating and purifying
b
trace analytes.
Minutes
Minutes
NEWPurification and
Isolation Cartridge Holders
9
Symmetry Capillary Columns
Symmetry columns provide the highest PicoFrit columns allow you to do femtomole peptide analysis. You can also spray directly from column to inlet
of a mass spectrometer and minimize post-column loss of sample.
standard of reproducibility with unmatched
peak symmetry for maximum sensitivity and
accurate quantification. IntegraFrit 75 m Columns
Polished edge
Distal end Polyimide coating Frit
IntegraFrit columns provide very low back pressure and are free of adhesives, binders and polymers. They can
be used with both ESI-MS and MALDI, and are Biomolecule and MS-compatible.
Method Development
10
Symmetry C18 PicoFrit Columns
Symmetry Columns
and Cartridges Dimension Particle Size Part Number
75 m x 50 mm 3.5 m 186001407
Symmetry Analytical Columns 75 m x 100 mm 3.5 m 186001408
75 m x 100 mm 5 m 186001409
75 m x 150 mm 5 m 186001410
Particle Part No. Part No.
Dimension Size C18 C8
1.0 x 50 mm 3.5 m WAT106056 WAT106052 Symmetry C18 IntegraFrit Columns
1.0 x 150 mm 3.5 m WAT248059 WAT248072
2.1 x 30 mm 3.5 m WAT058973 WAT058977
2.1 x 50 mm 3.5 m WAT200650 WAT200624 Dimension Particle Size Part Number
2.1 x 100 mm 3.5 m WAT058965 WAT058961 75 m x 50 mm 3.5m 186001403
2.1 x 150 mm 3.5 m WAT106005 WAT106011 75 m x 100 mm 3.5 m 186001404
3.0 x 100 mm 3.5 m 186000696 186000698 75 m x 100 mm 5 m 186001405
3.0 x 150 mm 3.5 m 186000695 186000697 75 m x 150 mm 5m 186001406
4.6 x 30 mm 3.5 m 186000271 186000270
4.6 x 50 mm 3.5 m WAT200625 WAT200620
4.6 x 75 mm 3.5 m WAT066224 WAT066200
4.6 x 100 mm 3.5 m WAT066220 WAT066204 Symmetry Column and Cartridge Column
4.6 x 150 mm 3.5 m WAT200632 WAT200630 Method Validation Kits
2.1 x 50 mm 5 m 186000206 186000212 Three Columns from Three Different Batches to Test Reproducibility.
2.1 x 150 mm 5 m WAT056975 WAT056955
3.0 x 150 mm 5 m WAT054200 WAT054230 Particle Part No. Part No.
3.0 x 250 mm 5 m 186000690 186000691 Dimension Type Size C18 C8
3.9 x 150 mm 5 m WAT046980 WAT046970
4.6 x 50 mm 5 m 186000207 186000213 4.6 x 150 mm Column 3.5 m WAT094240 WAT094237
4.6 x 150 mm 5 m WAT045905 WAT045995 2.1 x 150 mm Column 5 m WAT094234 WAT094231
4.6 x 250 mm 5 m WAT054275 WAT054270 3.0 x 150 mm Column 5 m WAT054446 WAT054434
3.9 x 150 mm Column 5 m WAT047210 WAT046955
4.6 x 150 mm Column 5 m WAT054448 WAT054435
Symmetry Cartridge Columns (Requires endfittings, see Page 13) 4.6 x 250 mm Column 5 m WAT054450 WAT054438
3.9 x 150 mm Cartridge* 5 m WAT054452 WAT054440
Particle Part No. Part No. 4.6 x 150 mm Cartridge* 5 m WAT054454 WAT054442
Dimension Size C18 C8 4.6 x 250 mm Cartridge* 5 m WAT054456 WAT054444
2.1 x 20 mm 3.5 m 186000269 186000268 * Require Endfittings, see Page 13
2.1 x 50 mm 3.5 m 186000152 186000149
2.1 x 100 mm 3.5 m 186000151 186000153
2.1 x 150 mm 3.5 m 186000150 186000148
4.6 x 75 mm 3.5 m WAT066260 WAT066210 SymmetryPrep Columns for Purification and Isolation
4.6 x 100 mm 3.5 m WAT066265 WAT066215
3.9 x 50 mm 5 m WAT054220 WAT054240 Particle Part No. Part No.
3.9 x 150 mm 5 m WAT054205 WAT054235 Dimension Size C18 C8
4.6 x 150 mm 5 m WAT054210 WAT054255 7.8 x 10 mm 5 m 186000711 186000712
4.6 x 250 mm 5 m WAT054215 WAT054245 7.8 x 50 mm 5 m 186000208 186000214
7.8 x 100 mm 5 m 186000209 186000215
19 x 10 mm 5 m 186000715 186000716
Symmetry Sentry Guard Columns (2 per pack)
19 x 50 mm 5 m 186000210 186000216
(Requires Sentry Guard Holders, see Page 13)
19 x 100 mm 5 m 186000211 186000229
30 x 50 mm 5 m 186000235 186000237
Particle Part No. Part No. 30 x 100 mm 5 m 186000236 186000238
Dimension Size C18 C8
7.8 x 10 mm 7 m 186000713 186000714
2.1 x 10 mm 3.5 m WAT106127 WAT106128 7.8 x 150 mm 7 m WAT066288 WAT066285
3.9 x 20 mm 5 m WAT054225 WAT054250 7.8 x 300 mm 7 m WAT066235 WAT066225
19 x 10 mm 7 m 186000717 186000718
19 x 150 mm 7 m WAT066240 WAT066228
Symmetry Capillary Columns 19 x 300 mm 7 m WAT066245 WAT066230
Purification and Isolation Cartridge Column Holders
Particle Part No. Part No.
Dimension Size C18 C8 7.8 x 10 mm 5, 7 m 186000708
19 x 10 mm 5, 7 m 186000709
0.32 x 50 mm 3.5 m 186000945 186000946
0.32 x 100 mm 3.5 m 186000951 186000952 Replacement o-ring 7.8 mm, 2 pkg 700001019
0.32 x 150 mm 3.5 m 186000957 186000958 Replacement o-ring 19 mm, 2 pkg 700001020
0.32 x 50 mm 5 m 186000929 186000930
0.32 x 100 mm 5 m 186000935 186000936
0.32 x 150 mm 5 m 186000347 186000941
11
SymmetryShield Column and Cartridge Column
SymmetryShield Method Validation Kits
Columns and Cartridges Three Columns from Three Different Batches to Test Reproducibility.
Particle Part No. Part No. 2.1 x 150 mm Column 3.5 m 186000182
Dimension Size RP18 RP8 4.6 x 150 mm Column 3.5 m 186000181 WAT094278
2.1 x 150 mm Column 5 m 186000100 WAT094254
1.0 x 50 mm 3.5 m 186000175 WAT106060 3.0 x 150 mm Column 5 m - WAT094251
1.0 x 150 mm 3.5 m 186000176 WAT106048 3.9 x 150 mm Column 5 m 186000104 WAT210594
2.1 x 30 mm 3.5 m 186000171 WAT106001 4.6 x 150 mm Column 5 m 186000103 WAT210588
2.1 x 50 mm 3.5 m 186000172 WAT094257 4.6 x 250 mm Column 5 m 186000102 WAT210591
2.1 x 100 mm 3.5 m 186000173 WAT058969
2.1 x 150 mm 3.5 m 186000174 WAT106008 3.9 x 150 mm Cartridge* 5 m 186000105 WAT210582
3.0 x 100 mm 3.5 m 186000700 186000703 4.6 x 150 mm Cartridge* 5 m 186000101 WAT210585
3.0 x 150 mm 3.5 m 186000699 186000702 4.6 x 250 mm Cartridge* 5 m 186000114 WAT210579
4.6 x 50 mm 3.5 m 186000177 WAT094260 * Require endfittings, see Page 13
4.6 x 75 mm 3.5 m 186000178 WAT094263
4.6 x 100 mm 3.5 m 186000179 WAT094266
4.6 x 150 mm 3.5 m 186000180 WAT094269
SymmetryShield Columns for Purification and Isolation
2.1 x 50 mm 5 m 186000217 186000223
2.1 x 150 mm 5 m 186000111 WAT094245
Particle Part No. Part No.
3.0 x 150 mm 5 m 186000692 WAT094243
Dimension Size RP18 RP8
3.0 x 250 mm 5 m 186000693 186000694
3.9 x 150 mm 5 m 186000108 WAT200655 19 x 10 mm 5 m 186001835 186001841
4.6 x 50 mm 5 m 186000218 186000224 19 x 50 mm 5 m 186001836 186001842
4.6 x 150 mm 5 m 186000109 WAT200662 19 x 100 mm 5 m 186001837 186001843
4.6 x 250 mm 5 m 186000112 WAT200670 19 x 150 mm 5 m 186001838 186001844
19 x 150 mm 7 m 186001839 186001845
SymmetryShield Cartridge Columns (Requires endfittings, see Page 13) 19 x 300 mm 7 m 186001840 186001846
Purification and Isolation Cartridge Column Holders
Particle Part No. Part No. 7.8 x 10 mm 5, 7 m 186000708
Dimension Size RP18 RP8 19 x 10 mm 5, 7 m 186000709
2.1 x 50 mm 3.5 m 186000168 186000147 Replacement o-ring 7.8 mm, 2 pkg 700001019
2.1 x 100 mm 3.5 m 186000167 186000146 Replacement o-ring 19 mm, 2 pkg 700001020
2.1 x 150 mm 3.5 m 186000166 186000145
4.6 x 75 mm 3.5 m 186000183 WAT094272
4.6 x 100 mm 3.5 m 186000170 WAT094275
3.9 x 50 mm 5 m WAT094248
3.9 x 150 mm
4.6 x 150 mm
5 m
5 m
186000106
186000110
WAT200658
WAT200665
Symmetry300
4.6 x 250 mm 5 m 186000113 WAT200661 Columns and Cartridges
SymmetryShield Sentry Guard Columns (2 per pack)
Symmetry300 Analytical Columns
(Requires Sentry Guard Holders, see Page 13)
12
Symmetry300 Cartridge Columns (Requires endfittings, see below) Symmetry300 Column and Cartridge Column
Method Validation Kits
Particle Part No. Three Columns from Three Different Batches to Test Reproducibility.
Dimension Size C18
2.1 x 50 mm 3.5 m 186000199 Particle Part No. Part No.
2.1 x 100 mm 3.5 m 186000191 Dimension Type Size C18 C4
2.1 x 150 mm 3.5 m 186000196
4.6 x 75 mm 3.5 m 186000192 2.1 x 150 mm Column 3.5 m 186000194 186000290
4.6 x 100 mm 3.5 m 186000193 4.6 x 150 mm Column 3.5 m 186000195 186000291
13
Sales Offices:
Austria and Export (Central and South
East Europe, CIS, Middle East, India
and India Subcontinent)
(43) 1 8771807
Australia (61) 2 9933 1777
Belgium and Luxembourg (32) 2 7261000
Brazil (55) 11 5543 7788
Canada 800 252 4752
China (86) 10 6526 0828
Czech Republic (42) 02 617 11384
Denmark (45) 46 598080
Finland (358) 9 506 4140
France (33) 1 30 48 72 00
Germany (49) 6196 40 06 00
Hong Kong (852) 2964 1800
Hungary (36) 1 350 5086
India (91) 80 8371900
Ireland (44) 208 238 6100
Italy (39) 02 27421.1
Japan (81) 3 3471 7191
Malaysia (60) 3 704 8600
Mexico (5255) 5524 76 36
The Netherlands 31 (0) 76 508 72 00
Norway (47) 638 46 050
Poland (48) 22 883 4400
Puerto Rico (787) 747 8445
Russia/CIS (7) 095 931 9193
Singapore (65) 278 7997
Spain (34) 93 600 93 00
Sweden (46) 8 594 69 200
Switzerland (41) 62 889 2030
Taiwan (886) 2 543 1898
United Kingdom (44) 208 238 6100
All other countries:
Waters Corporation U.S.A.
508 478 2000/800 252 4752
Literature Code
Symmetry Application Notebook WN033
SymmetryShield Application Notebook WN035
SymmetryShield White Paper
720000207EN
To request your free copy, or for more information,
please contact your local Waters Sales Office.
The quality management system of Waters manufacturing facility, Taunton, Massachusetts, and
Wexford, Ireland complies with the International Standard ISO 9002 Quality Management and
Quality Assurance Standards. Waters quality management system is periodically audited by the
registering body to ensure compliance.
2002 Waters Corporation. Symmetry, Symmetry300, SymmetryShield, SymmetryPrep, Alliance and Waters
are trademarks of Waters Corporation. IntegraFrit, PicoFrit, and PicoTip are trademarks of New Objectives.
www.waters.com
720000454EN Online Storeclick on c.shop