Chromatography Columns and Supplies Catalog www.waters.com

Transcription

Chromatography Columns and Supplies Catalog www.waters.com
Chromatography
Columns and
Supplies Catalog
2001–2002
www.waters.com
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Waters Chromatography Chemical Manufacturing:
"In a Class by Itself"
by Frederick G. Harrison, Director, Manufacturing Operations
Waters manufactures its consumable chromatography products in Taunton, Massachusetts and in a recently opened facility
in Wexford, Ireland. Wexford is located 90 miles south of Dublin on Ireland's southeast coast. Our manufacturing operations
continue to be unique as compared to others in the industry. Our primary mission is to synthesize high-quality chromatography supports, pack chromatography columns, assemble solid-phase extraction devices, and to produce reagent kits. Since
some of our products are utilized as compounds in medical devices, our facilities are registered with the U.S. Food and Drug
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Registration number 100,238.
The 33,000 sq. ft. Taunton, Massachusetts facility, built in 1978, is actually two buildings in one: an explosion proof
chemical processing area, and the rest of the facility which includes specialty column packing and solid-phase extraction
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Our brand new two-story 25,000-sq. ft. Wexford, Ireland facility sits on 4 acres of land. This facility manufactures the bulk of
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goods stores, administration offices, and a bunkered warehouse.
Chemical Processing
The chemical processing section of the Taunton facility manufacturers over 200 different silica or polymer-based bulk
chromatographic materials and reagents including XTerra™, Symmetry®, Nova-Pak®, µBondapak™, and Waters Spherisorb®
products. Chromatographic particles are first made and then sized using air or mechanical classification. We then chemically
bond various ligands to the base particle to provide you with a range of selectivity, such as C18, C8, phenyl, and so forth.
These reactions take place in glass-lined Pfaudler reactors, ranging in size from 5 gallons to 100 gallons. These bonded
particles are then isolated from the reaction mixtures and dried. Throughout this entire process, highly skilled chemical
technicians follow detailed operating procedures very much like those used in the pharmaceutical industry. After each step,
technicians indicate completion of a given operation by recording the time, date, and their initials on batch records. These
processes are also monitored in our quality laboratory. Samples are taken at critical points in the operation and results are
then analyzed and compared to in-process specifications. Finally, the finished bulk packing is tested in the quality control
laboratory both chemically and chromatographically to ensure that it meets the same level of performance as previously
manufactured material--giving you the same reproducibility each time you use a Waters’ chromatographic product.
Column Packing and Solid-Phase Extraction Assembly at Both Facilities
To provide you with the highest quality chromatographic product, skilled technicians carefully pack over two thousand
different steel columns, polyethylene cartridges, solid-phase extraction cartridges and reagent kits according to detailed
operating procedures, like the ones described earlier.
Once the columns are packed, they are chromatographically tested to ensure quality and consistency using state-of-the-art
equipment. Solid-phase extraction devices are tested for backpressure, flow rate, and in some cases, extraction efficiencies.
Each column or lot of extraction devices is then labeled with a serial number and packaged. This serial number provides
traceability for every column and device lot and documents complete batch information for the packing material used in the
finished product. We can tell you when it was tested, who tested it, and the results of the test for a chromatographic product
manufactured 3 months or 3 years ago. This documentation guarantees that when you purchase your next Waters column or
device, it will perform the same way as your last column or device.
We are proud of our manufacturing facilities and invite you to visit us.
Tours of either site can be arranged by your Waters Technical Representative.
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Waters warrants that all products sold by them will be of good quality and
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will Waters be liable for consequential, economic or incidental damages of any nature.
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or product.
All prices are subject to change without notification.
Trademark Acknowledgements
Trademarks of Waters Corporation
Accell, AccQ-Fluor, AccQ•Tag, AccuPure, AccuSep, Alliance, Bondapak, µBondapak,
CapLC, C-Gel, Delta-Pak, EnantioSelect, Envirogel, Gen-Pak FAX, Guard-Pak, IC-Pak,
IonSelect, Nova-Pak, Oasis, Pico•Tag, Porapak, Porasil, µPorasil, PrepLC, PrepPak,
Protein-Pak, Radial-Pak, Resolve, SAM, Sentry, Sep-Pak, Silica-Pak, Styragel, µStyragel,
Sugar-Pak, Symmetry, SymmetryPrep, SymmetryShield, Symmetry300, Ultrahydrogel,
Ultrastyragel, XPoSure, Waters Spherisorb, XTerra, c.shop and Waters
All other trademarks in the catalog are acknowledged.
Acrodisc
AcroPrep
Cheminert
Florisil
FPLC
Inter-Seal
µPage
Partisil
Shodex
Slip-Free
Supor
Z-Spin
Pall Gelman Laboratory
Pall Gelman Laboratory
VICA
US Silica Corporation.
Pharmacia LKB
Integrated Liner Technologies, Inc.
J&W
Whatman
Showa Denko
Keystone Scientific, Inc.
Pall Gelman Laboratory
Pall Gelman Laboratory
Click anywhere on Table of Contents; Type desired page number; Click on OK.
Sample Preparation
Page
Solid-Phase Extraction
Sample Preparation
Introduction......................................................................................6
Speciality Products .........................................................................7
Oasis® Sample Extraction Products......................................................7
Porapak® RDX Sep-Pak® Extraction Cartridge.......................................24
Sep-Pak® DNPH-Silica Cartridges .....................................................22
Waters XPoSure™ Aldehyde Sampler Cartridge ..................................23
Ordering Information
Oasis® Sample Extraction Products ...........................................20
Speciality Cartridges ..............................................................25
Waters Solid-Phase Extraction Applications Guide
and Bibliography, 6th Edition ..................................................26
Sep-Pak Cartridges and Accessories .............................................26
General Elution Protocols on Sep-Pak® Cartridges ...............................33
Sep-Pak® Cartridge Selection Guide .................................................34
Which Cartridge is Best for Your Application .....................................27
Which Sorbent is Best for Your Application ........................................29
Ordering Information
Sep-Pak® Cartridges...............................................................36
Waters Extraction Manifold and Accessories .............................40
Sample and Solvent Filtration
Alliance® Filtration System ................................................................42
Ordering Information
AcroPrep™ Filter Plates for the Alliance® Filtration System...............43
AcroPrep™ Vials, Accessories and Spare Parts............................43
Pall Gelman Laboratory Sample and Solvent Filtration Products
Introduction ...........................................................................44
Filter Design and Membrane Choices.......................................45
Part Number Cross Reference..................................................46
Ordering Information
Solvent Filtration Membranes and Apparatus .............................50
Syringe Filters ........................................................................47
5
Sample Preparation
Introduction
Sample preparation is a key procedure in modern chemical analysis.
By some estimates, 60-80% of the work activity and operating cost in an
analytical laboratory is spent preparing samples for introduction into an
analytical system or device.
The importance of sample preparation stems
from three major concerns:
Sample Concentration
Frequently, the component of interest is present in levels too low for detection. Sample preparation can concentrate the component to necessary
levels for accurate and precise measurement.
Contaminants
The presence of interfering matrix elements can mask or interfere with the
analysis of the component(s) of interest. Sample preparation can remove
excess contaminants to yield clean, informative chromatograms.
Strategies for Solid-Phase Extraction
There are two simple solid-phase extraction strategies for sample preparation. You can choose a cartridge sorbent and sample solvent to cause the
component(s) of interest to be:
1. unretained while matrix interferences are adsorbed
2 retained while matrix interferences pass through unretained
The first strategy is usually chosen when the desired sample component is
present in high concentration. When components of interest are present at
low levels, or multiple components of widely differing polarities need to
be isolated, the second strategy is generally employed. The second
strategy may also be used for trace enrichment of extremely low level
compounds and concentration of dilute samples. A complex matrix may
be treated by both elution strategies to isolate different target analytes.
With either strategy, there are three different chromatographic modes to
choose from.
You can select :
In Solution
For most analyses (HPLC, GC, Spectrophometry, RIA, etc), the sample
must be properly prepared in solution for subsequent analysis.
• normal phase
Common methods for sample preparation include liquid-liquid extraction,
centrifugation, and solid-phase extraction. Solid phase extraction (SPE) is
one of the simplest, yet most effective and versatile, methods of sample
preparation.
• ion-exchange
Utilizing low cost, prepacked, disposable cartridges, a sample component of interest is separated from other species by applying the sample
mixture to a solid chromatographic sorbent and selectively eluting the
desired component(s).
Convenient, Easy-to-Use
Convenience and ease-of-use are primary benefits of Sep-Pak® solidphase extraction cartridges, to maximize the productivity of your analytical
operation while saving you time and money. When compared to other
sample preparation techniques, SPE cartridges also offer:
Faster sample prep — average time cut by 2/3
Lower cost — less solvent and reagent consumption and less
hazardous waste for disposal
• reversed-phase
The basic characteristics of each chromatographic mode are described
on page 25. There are many different types of sorbents for each mode
and the selection of strategy, mode, sorbent, and elution solvents will
depend upon the specific sample mixture and goal of the separation.
For example, the first strategy may be used prior to the analysis of organic acids in red wine. By passing the wine sample at the appropriate pH
through a C18 reversed-phase cartridge, the interfering wine pigments are
removed by adsorption while the organic acids elute unretained.
With a similar sample but a different goal, the second strategy can be
used to isolate phenolic pigments in red wine, also with a C18 cartridge.
In this case, the sugars and acids pass directly through the cartridge,
while the pigments are retained and subsequently eluted with a stronger
mobile phase. Of course, it is possible to combine two such applications
into a single, multi-step protocol to prepare several classes of compounds
for analysis.
Greater recoveries — minimal sample transfer
Greater accuracy — no cross contamination
Less sample handling — no emulsion problems
Reduced harm to labile samples — minimal
evaporation
Improved safety — due to reduced solvent/sample exposure
and glassware
Easy automation — simultaneous batch processing of
multi-samples
© 2001 Waters Corporation
6
Sample Preparation
Oasis® Family of HLB, MCX & MAX Sample Extraction Products
Sample Preparation
Setting New Standards for Solid-Phase
Extraction (SPE) Technology
The extensive offering of the Oasis® sample extraction products allows
you to select the device with the sorbent amount that meets your assay
requirements.
Available chemistries:
HLB (Hydrophilic-Lipophilic Balance) reversed-phase sorbent.
•
Three Polymeric Sorbents, pH Stable 0-14
•
High and Consistent Recoveries
•
Unmatched Batch-to-Batch Reproducibility
•
Simple and Fast SPE Method Development
•
1 – 35 cc Sample Extraction Cartridges with
HLB, MAX, MCX Sorbents
•
96-Well Extraction Plates with HLB, MCX
and MAX Sorbents for High Throughput
LC/MS/MS
•
Sample Extraction Columns and Prospekt™
Cartridges with HLB Sorbent
•
HLB Glass Sample Extraction Cartridge
•
Bulk and Other Products available Upon Request
MCX (Mixed-mode: Cation-eXchange and reversed-phase) sorbent.
MAX (Mixed-mode: Anion-eXchange and reversed-phase) sorbent.
Current Oasis Patents:
®
Patent No. 5,882,521 (1996), Patent No. 5,976,376 (1998)
Patent No. 6,106,721 (1999), Additional Patents Pending
© 2001 Waters Corporation
7
Sample Preparation
Oasis® HLB plates, cartridges and columns
High and Consistent Recoveries
Oasis® sorbents are water-wettable maintaining high retention and capacity for a wide spectrum of analytes, especially when the SPE column runs
dry. Oasis® sorbents retain polar compounds far better than bonded silica
SPE sorbents, figure 2.
High Capacity
When transferring methods from a C18-bonded phase to Oasis® products, keep in mind the greater capacity of the Oasis® sorbent. The Oasis®
sorbent has 2-3x more surface area and shows a dramatic increase in
k values compared to silica-based C18, figure 3.
Figure 2: Effect of Drying on Recovery - Oasis® HLB
versus C18 Sorbents. No Stopcocks Required!
C18 Cartridge (100 mg)
Oasis HLB Cartridge (30 mg)
®
100
A Breakthrough in SPE
100
80
80
In October 1977, Waters designed the first miniature cartridge columns
(Sep-Pak® cartridges) containing silica-based adsorbents for SPE.
New demands for sample preparation led to the development of a new,
specially designed polymeric sorbent which performs optimally for
reversed-phase SPE. The Oasis® HLB copolymer with unique HydrophilicLipophilic Balance (figure 1) is unlike traditional SPE sorbents.
Today's goals for modern solid-phase extraction (SPE) are faster throughput, higher recovery and reproducibility, stronger retention and selectivity.
Now SPE can outpace high throughput techniques such as LC/MS/MS.
% Recovery
60
Introduction
40
20
% Recovery
Procainamide
Acetaminophen
Ranitidine
Propranolol
Doxepin
60
40
20
0
0
0
5
10
0
Drying Time (minutes)
4
Oasis® HLB 1cc/30 mg and 1cc/100 mg C18 cartridges were conditioned on a Waters
vacuum manifold. When the methanol reached the top of the upper frit in each cartridge,
vacuum was maintained for different times to vary the cartridge drying time. The SPE protocol
was then continued. The data shown are the average of three replicate extractions.
Figure 1: Unique Water-Wettable HLB Copolymer
O
Figure 3: Higher Retention Means Greater
Capacity, No Breakthrough
N
Retention Factor (k ) Comparison
C
18
H O
O
N
N
O
H
Divinylbenzene
N
O
CH 3
N
H3 C
Acetaminophen
OH
HO
O
HO
Optimal Properties for Reversed-Phase SPE
Specific Surface Area: 810 m2/g
Average Pore Diameter: 80 Å
Total Pore Volume: 1.3 cm3/g
Average Particle Diameter: 30 µm or 60 µm
50
100
Data shown were obtained with two 3.9 mm x 150 mm columns, each packed with one
of the sorbents, operated under the same conditions: mobile phase: 20 mM potassium
phosphate, pH 7.0/methanol (95/5 v/v); temperature: 30 °C; flow rate: 1.0 mL/min;
detection: UV @ 254 nm.
© 2001 Waters Corporation
8
Catechol
0
The Oasis® HLB sorbent is a macroporous copolymer made from a balanced ratio of two
monomers, the lipophilic divinylbenzene and the hydrophilic N-vinylpyrrolidone.
Oasis® HLB Cartridge
Theobromine
N
N
CH 3
H
N
Hydrophilic – Lipophilic Balance
Cartridge
Salicylic acid
HO
O
O
N-Vinylpyrrolidone
8
Drying Time (minutes)
Sample Preparation
Exceptional Batch-to-Batch Reproducibility
Figure 4: General SPE Method (1-D)*
The exceptional batch-to-batch reproducibility of the Oasis® sorbent
eliminates your need to reserve batches of sorbent for your SPE work.
Condition
1 mL methanol
Sample Pretreatment
Suggestion
Applying one or more of the
following steps before loading
your sample may improve
your results:
Equilibrate
1 mL water
Load
1 mL spiked sample
A Simple and Fast Generic (1-D) SPE
Protocol for Rapid Method Development
• Dilute sample 1:1 with buffer
to improve flow during loading
Wash
1 mL 5% methanol in water
Oasis® HLB sorbent makes it fast and easy to develop sample preparation methods that deliver high, reproducible recoveries especially
suited to LC/MS/MS analysis by providing the required selectivity
and sensitivity.
Unlike silica-based sorbents where you must evaluate several different
cartridges, acidic, basic, and neutral compounds, whether polar or
nonpolar, can be isolated reproducibly (RSDs <5%) with high recovery
(>85%), using the same simple SPE protocol, figures 4 and 5. The
generic scheme, figure 4, which has proven useful for a wide variety of
compound types may be the only protocol required, reducing method
development time.
Sample Preparation
As a result of careful process design and stringent quality controls, a new
standard has been set in batch-to-batch and lot-to-lot reproducibility for
SPE methods.
• Add 20 µL (H2PO4) to disrupt
drug protein interaction
Elute
1 mL methanol
• Dilute 1:1 or greater with
0.1 N HCL or other acids
Evaporate & Reconstitute
40 ˚C/under nitrogen stream
200 µL mobile phase
• Filter through 0.45 µm membrane
• Centrifuge @ ≥3000 rpm
* Volumes are given for the Oasis®
HLB cartridge 1cc/30 mg
Figure 5: One Simple Procedure: Many Applications on a Universal Sorbent
100
80
sed sorbents
With silica-ba
ral
uld need seve
wo
u
yo
)
(C18
ies to achieve
istr
em
ch
t
en
differ
n here
the results show
60
■
■
■
% Recovery
Acids
Neutrals
Bases
n=6
40
RSDs < 3.5%
20
e
m
er
az
in
e
in
op
Th
he
eo
n
br
om
Pa
i
n
ra
za e
nth
Th
i
n
eo
e
ph
yll
in
e
C
af
Pr
oc fein
e
ai
na
m
id
e
Ra
ni
tid
O
in
e
xy
co
do
Pr
op ne
ra
no
N
lo
l
al
tre
x
Sa one
lb
uta
m
o
Do l
xe
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n
id
in
ta
m
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0
Seventeen drugs with a wide range of polarities were spiked in porcine serum, then
extracted with 1cc/30 mg Oasis HLB cartridges, using the same SPE method, figure 4.
20 µL/mL H2PO4 was used to disrupt drug-protein binding.
®
© 2001 Waters Corporation
9
Sample Preparation
Solvent and Sorbent Amount Selection
for the Generic Method
The amount of sorbent in a cartridge or a plate required to meet your
application, is given in Table 1. The solvent used for the elution step
should be selected based on the polarity of the analyte. Table 2 gives a
selection of elution solvents and each solvent gives you different selectivity
and elution strength.
Table 1: Capacity and Elution Volume of Oasis®
96-Well Plates and Cartridges
Sorbent
per device
Maximum
Mass Capacity
Typical
Sample Volumes
Elution
Volume
0.15 to 1 mg
0.35 to 2 mg
1 to 5 mg
2 to 10 mg
10 to 100 µL
50 to 200 µL
100 µL to 1mL
200 µL to 2 mL
≤150 µL
≤250 µL
≥400 µL
≥800 µL
5 mg*
10 mg
30 mg
60 mg
* Available only in 96-well plates
When converting from C18 silica-based to Oasis® SPE sorbents, use approximately 1⁄3 less Oasis®
sorbent. For example 100 mg C18 sorbent = 30 mg Oasis® sorbent
Table 2: Tips for Selecting Elution Solvents for the Generic
SPE Method (1-D)* The elution solvent is selected based
on polarity of analyte.
Solvent
Solvent Type
Relative
Elution Strength**
Methanol
Acetonitrile
Tetrahydrofuran
Acetone
Ethyl Acetate
proton donor
dipole-dipole
dipole-dipole
dipole-dipole
dipole-dipole
1.0
3.1
3.7
8.8
high
Methylene Chloride
dipole-dipole
high
Comments
disrupts H-bonding
medium polarity drugs
medium polarity drugs
medium polarity drugs
nonpolar drugs and
GC compatible
nonpolar drugs and
GC compatible
* When using solvents other than methanol, add 10-30% methanol to disrupt
H-bonding on the Oasis® HLB sorbent.
** High-Purity Solvent Guide. Burdick & Jackson Laboratories, Inc. Solvent Properties of Common
Liquids, L.R. Snyder, J. Chromatogr., 92, 223 (1974); J. Chromatogr. Sci. 16, 223 (1978)
Automation
Automation of Oasis® Sample Extraction Cartridges and 96-Well Plates
Oasis® SPE products are compatible with the following
liquid handling and/or SPE automation systems
• Packard MultiProbe II® Robotic liquid
handling system
Alliance® HT System
for LC/MS(MS)
• Tomtec Quadra96® and Quadra96®
SV Pipetting Station
• Control via Micromass MassLynx
mass spectrometry software
• Hamilton Microlab® SPE Workstation
• Beckman Biomek® 2000 Laboratory
Automation Workstation
• Fast cycle times for high throughput
with 96-well plates
• Zymark RapidTrace® Automated
SPE Workstation
• Assay integrity assured with sample
manager's temperature control and
very low carryover performance
• Gilson ASPEC XL4
• Compatibility with the Oasis®
on-line extraction columns
• Gilson ASPEC XL
• Gilson 215 SPE Liquid Handler
• Tecan Genesis
• Prospekt™/Prospekt™ II
© 2001 Waters Corporation
10
Sample Preparation
Low Elution Volume and High Throughput
96-Well Extraction Plates
Figure 7: Comparisons to Oasis® Low Elution Volume Plate
Sample Preparation
100
The Oasis® sorbent, with different amounts in a single 96-well plate,
figure 6, gives high and reproducible recoveries for a wide range of
analytes including polar and basic drugs, with RSDs ≤ 5% (n=96).
Waters offers the standard high-throughput 30 mg and 60 mg Oasis®
96-well plates and has now introduced the 5 mg and 10 mg 96-well
low elution plates.
80
% Recovery
60
Eliminate Evaporation Step With Low Elution
Volumes Without Sacrificing Capacity
40
2
The high capacity of the Oasis® sorbents makes plasma sample preparation with low elution volumes possible without sacrificing performance,
even for highly polar drugs and metabolites, figure 7. Eluting drugs and
their metabolites in 150 µL or less volume, eliminates the time consuming
and tedious evaporation and reconstitution steps saving hours of assay
time. Just elute and shoot. This makes Oasis® products ideally suited for
situations where high-throughput SPE is a requirement.
0
Oasis 10 mg Plate
®
C18 Disk Plate
Procainamide
Theophylline
Acetaminophen
Dipropyl Phthalat
100
Figure 6: Waters 96-Well Plate Design
2000
1R&
9
0D
Rin&
Aw
ar9dD9W10
ner
80
% Recovery
100 Award
Two- age
Selected as one St
of the 100 most
technol
ogically es
W
significa
nt
proel
ig
duclts D
of the yea
r n
by R&D Magazine
60
40
2
0
Oasis HLB
®
Polymer A
Polymer B
Procainamide
5 mg
10 mg
30 mg
60 mg
Ranitidine
The Oasis HLB copolymer in the 96-well plate gave significantly higher recovery for
polar drugs than two traditional polymeric materials, or a silica-based sorbent.
®
By varying frit size and/or placement, the same plate may be filled with various
quantities of sorbent per well. Our design permits optimal recoveries, even with
low sorbent weights for smaller elution volumes.
Low Elution Volume Oasis® HLB Extraction Plate,
5 mg/well, Generic SPE Procedure
Condition/Equilibrate
0.5 mL methanol/0.5 mL water
Load
100 µL of spiked plasma
Wash 1
0.5 mL 5% methanol
Elute
100-150 µL 70% acetonitrile/
30% methanol
TM
Visit Waters on-line
product resource today
Dilute
1:1 to 3:1 with water/mobile phase
© 2001 Waters Corporation
11
Sample Preparation
Selective 2-Dimensional
(2-D) Method Development
Figure 8: Retention Factor Versus pH
for Acids, Bases and Neutrals
Meet Selectivity and Sensitivity Goals
20
Theory
18
The generic method can be expanded to meet selectivity and sensitivity
goals. A more specific method can be developed to remove plasma or
urine interferences. A cleaner extract is achieved by simply manipulating
the organic concentration and/or the pH.
Acids (HA)
16
B
Retention Factor
14
Figure 8 demonstrates how a base compound is ionized at low pH
(2 units below its pKa), therefore poorly retained by SPE, and at high
pH (2 units above its pKa), strongly retained by reversed-phase SPE.
(pKa = 4.8 )
Neutrals
12
(pKa = 9.0 )
10
8
6
4
A Wash Elute Study is performed to determine the optimal SPE method
using organic concentration and pH.
A-
Bases (BH+)
2
0
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
pH
The 2-D experiment in figure 9 A and B demonstrates this washing, with
70% base modified MeOH in H2O or 30% acid modified MeOH in
H2O, will not remove analyte from cartridge. To facilitate complete elution
of analyte, an acidified (2% acetic acid in 65% methanol) can be
utilized, figure 10.
Recommended pH range of Silica
Recommended pH range of Oasis®
Figure 9A: 2-D Wash Elute Study: Experiment for Oasis® HLB 30 mg 96-well Plate
Prepare analytes in saline
▼
From these results, the wash solvent(s) can be either
acidic, up to 30% methanol containing 2% acetic
acid, or basic, up to 70% methanol containing 2%
ammonium hydroxide, since no analyte is eluted under
these conditions. To obtain complete elution, the elution
solvent should be acidic (2% acetic acid) and the
methanol concentration should be greater than 60%.
Condition/equilibrate 20 cartridges
1 mL methanol/1 mL water
▼
Load 1 mL spiked saline solution
▼
Elute with 1 mL
Peak Area (µVs)
2% NH4OH in MeOH/water
Verapamil
Norverapamil
Methoxyverapamil
0-70%
No Response
No Response
No Response
80%
34,095
47,482
6,358
90%
63,158
85,704
178,178
0% 10% 20% 30% 40% 50% 60% 70% 80% 90%
Peak Area (µVs)
0-30%
Verapamil
Norverapamil
Methoxyverapamil
No Response
No Response
No Response
40%
73,146
32,269
86,789
50%
121,820
68,259
137,899
60%
109,145
61,830
123,229
200000
Base Modified Wash
150000
100000
50000
0
Acid Modified Wash
150000
Maximum %
Methanol Wash
100000
50000
0
20
40
60
80
% MeOH
© 2001 Waters Corporation
12
0% 10% 20% 30% 40% 50% 60% 70% 80% 90%
Figure 9B: 2-D Wash Elute Study Results
Area (µV*sec)
Acid 2%
Acetic Acid
2% CH3COOH in MeOH/water
Area (µV*sec)
Base 2%
Ammonium
Hydroxide
100
Minimum %
Methanol
Elution
0
0
20
40
60
% MeOH
80
100
Sample Preparation
Selective 2-Dimensional (2-D) Method
Development (continued)
Figure 10: 2-D SPE Method Developed from Wash Elute Study
utral
Removes ne
d basic
non-polar an
erferences
lipophilic int
Equilibrate
1 mL water
Wash 3
1 mL 65% MeOH in water
with 2% ammonium hydroxide
Load
1 mL spiked sample
sic
Elute the ba
in
compounds
ionized form
utral
Removes ne
sic
polar and ba
ences
polar interfer
Wash 1
1 mL 5% MeOH in water
with 2% acetic acid
idic
Removes ac
ences
polar interfer
Wash 2
1 mL 5% MeOH in water
with 2% ammonium hydroxide
Elute
1 mL 65% MeOH in water
with 2% acetic acid
Evaporate & Reconstitute
40 ˚C/under nitrogen stream
200 µL water/mobile phase
Figure 11: Better Results with a 2-Dimensional Method
0.004 AU
1
High
2
AV
100%
pH
3
Sample
3
2
Column:
5%
Blank
1
Low
Low
Organic
Concentration
High
2-D Method with 2 washes
AV
1
High
2
0.004 AU
3
65%
2
pH
3
Sample
Blank
100%
SymmetryShield™ RP8,
5 µm, 3.9 x 150 mm
Temperature:
30 °C
Mobile Phase:
50 mM Phosphate pH 7:
acetonitrile:methanol
(41:37:22); Alliance® 2690
Detection:
UV at 230 nm
Flow Rate:
1.0 mL/min.
Injection Volume: 40 µL (after evaporation and
reconstitution in 200 µL water)
Peak Identification:
Peak 1: Norverapamil
Peak 2: Verapamil
Peak 3: Methoxyverapamil (I.S.)
5%
1
4
Low
Low
Organic
Concentration
High
2-D Method with 3 washes
2
1
High
3
0.004 AU
AV
3
pH
1
Low
Low
0
2
4
6
8
100%
5%
Sample
Blank
4
65%
2
5
Organic
Concentration
High
10
Minutes
© 2001 Waters Corporation
13
Sample Preparation
Condition
1 mL methanol
A cleaner extract was achieved for verapamil and its metabolite,
figure 11, using the 2-D procedure, figure 10, instead of the generic
method in figure 4. The more selective method enables higher sensitivity
for analyses at ng/mL levels.
Sample Preparation
Oasis® MCX plates, cartridges and columns
Figure 13: Oasis® MCX Method for Extraction of Basic Compounds
ning
No conditio
Load
3 mL spiked and acidified urine
Wash 11
Wash
2 mL 0.1N HCI
utral and
Removes ne
ences
acidic interfer
Wash 2
2 mL methanol
Elute B
2 mL 5% ammonium hydroxide
in methanol
Evaporate and Reconstitute
300 µL 20% methanol in water
Oasis® MCX 3 cc/60 mg cartridge
High Selectivity and Sensitivity for Basic
Compounds With No Conditioning of the
Oasis® MCX Sorbent
Figure 14: Comparison of Oasis® MCX Cartridges to other SPE
Silica-Based Products Using the Generic Oasis® MCX Method
100
Obtain selective retention of basic drugs with cation-exchange groups.
The Oasis® MCX sorbent has a tightly controlled ion-exchange capacity
(1 meq/gram), figure 12. There are no silanol groups to complicate
the retention mode or method development. This novel, water-wettable,
polymeric sorbent is stable from pH 0 to 14, making method
development simple and fast.
Methadone
40
Propranolol
20
0
The use of a pH modified wash system allows optimization of the method
and it produces a very selective protocol for basic compounds, figure 15.
13%
12%
1.1%
Methadone metabolite
(EDDP)*
2.4%
3.2%
3.2%
As shown in figure 13, one protocol with minimum wash steps gives
extractions fast enough to keep pace with your analytical system. No
conditioning or equilibration of cartridges is required to achieve excellent
results for basic drugs, see figure 14.
60
0.3%
0.7%
0.5%
% Recovery (% RSD)
80
Oasis® MCX
60 mg/3 cc
Brand A
300 mg/
3 cc
Brand B
300 mg/
3 cc
Figure 15: Selectively Separate Primary and Secondary
Amines from Tertiary and Aromatic Amines Using an
Oasis® MCX Cartridge
Figure 12: Oasis® MCX Sorbent
100
O
N
Toluidine (Aromatic)
75
Doxepin ( 3˚)
Amitriptyline ( 3˚)
SO3 H
SO3 H
Procainamide ( 3˚)
85
Elute tertiary and aromatic
amines with 5% TEA
50
Elute primary and secondary
amines with 5% NH4OH in methanol
25
HO3S
N
0
Mixed-Mode, Cation-Exchange Reversed-Phase Sorbent
N-methylbenzylamine ( 2˚)
Benzylamine (1˚)
Methamphetamine ( 2˚)
0
200
400
600
800
Elution Volume (µL of 5% TEA in Methanol)
© 2001 Waters Corporation
14
N,N-dimethylbenzylamine ( 3˚)
Amphetamine (1˚)
15
O
Nortriptyline ( 2˚)
1000
Sample Preparation
Oasis® MAX plates, cartridges and columns
Figure 16: Oasis® MAX Sorbent
Sample Preparation
O
N
+
CH2 NR 3
+
CH 2NR3
N
+
R3NCH2
O
Mixed-Mode, Anion-Exchange Reversed-Phase Sorbent
Figure 17: Generic Oasis® MAX Method
for Extraction of Acidic Drugs
High Selectivity and Sensitivity for Acidic
Compounds using the Oasis® MAX Sorbent
Condition
1 mL methanol
The Oasis MAX sorbent, figure 16, has a tightly controlled ion exchange
capacity of 0.3 meq/gram ensuring reproducible SPE protocols for
extraction of acidic compounds and metabolites from biological fluids.
®
This novel, water wettable polymeric sorbent is stable from pH 0 to 14
making method development simple and fast, figure 17. As shown in
figure18, acidic compounds can be separated from basic and neutral
compounds on the same Oasis® MAX cartridge.
sic and
Removes ba
ferences
neutral inter
Equilibrate
1 mL water
Wash 2 (Elute 1)
2 mL methanol
Load
3 mL sample
Elute 2
2 mL 100 mM phosphoric
acid/acetonitrile 1:1 (v/v)
Wash
2 mL 50 mM sodium
acetate pH 7/5% methanol
Alternative LC/MS Elution System:
2% formic acid in methanol
Figure 18: High Recovery of Acidic, Basic and Neutral Compounds Using the Generic Oasis® MAX Method
Elute 1 (basic and neutral drugs)
Elute 2 (acidic drugs)
1
2
I.S.
I.S.
AV
AV
0
2
4
6
8
10
Sample
Sample
Blank
with I.S.
Blank
with I.S.
0
2
Minutes
Column:
4
3
SymmetryShield™ 3.5 µm, 4.6 mm x 100 mm
SymmetryShield™ Guard column, 5 µm,
3.9 mm x 20 mm
Mobile Phase:
20 mM potassium phosphate,
pH 2.7/methanol, 52:48 v/v
Detector:
UV @ 214 nm
Temperature:
30 °C
Flow Rate:
2 mL/min
Injection Volume: 10 µL of extract with internal standard
4
6
8
10
Minutes
Peak Identification
1. Nortriptyline
2. Secobarbital
3. Ketoprofen
4. Naproxen
I.S. Butylparaben
Results (n=6)
Drugs
Nortriptyline
Secobarbital
Ketoprofen
Naproxen
µg/mL
Type of Drug
% Recovery
% RSD
5
10
5
2.5
Basic (Elute 1)
Neutral (Elute 1)
Acidic (Elute 2)
Acidic (Elute 2)
94.34
88.53
91.64
103.43
0.78
1.03
2.35
3.04
1 mL urine hydrolyzed (1M KOH 60 °C, 15 minutes) and adjusted to pH 2
with phosphoric acid; diluted 1:1 with 10 mM sodium acetate, pH 2.
© 2001 Waters Corporation
15
Sample Preparation
Ultrafast On-Line SPE With Oasis®
Extraction Columns for LC/MS/MS
Details of the technique
Ultrafast chromatography is a robust and straightforward technique that
combines HPLC and mass spectrometry.
A diluted plasma sample is injected onto the Oasis® HLB 1 mm x 50 mm
column. The column is then purged of most protein and interferences.
A fast gradient is run to elute the target analyte and one tenth of this
flow (400 µL/min) is diverted to the mass spectrometer for detection,
figure 18.
The Oasis® HLB 2.1 mm x 20 mm cartridge column is very rugged and
gives even more robust assays and higher sensitivity. Up to 360 injections
were achieved on a single column without any visible deterioration of the
peak shape, figure 20.
Oasis® Cartridge Column
2.1 mm x 20 mm
Figure 18: Ultrafast, on-line chromatography
Oasis® HLB cartridge column
1 mm x 50 mm
PUMP
Ultrafast on-line analysis of multiple analytes can be accomplished using
the Oasis® extraction column, combined with a traditional analytical
column, as shown in figure 19.
Waste
Switching
Valve
INJ
4 mL/min
Flow Splitter
10:1
Oasis® Column
1 mm x 50 mm
400 µL/min
MS/MS
Selected
Reaction
Monitoring
Ayrton, J, Dear GJ, Leavens WJ, Mallett DN, Plumb RS.
Optimization and routine use of generic ultra-high flow-rate liquid chromatography with
mass spectromic detection for the direct on-line analysis of pharmaceuticals in plasma.
J Chromatogr A. 1998 Dec 18;828(1-2):199-207
Figure 19: Fast on-line analysis: Multiple drugs with dual columns
Oasis® HLB cartridge column
2.1 mm x 20 mm
PUMP
Figure 20: Column Lifetime: Overlay of Toluamide
after 360 Plasma Injections
Waste
Column:
Mobile Phase:
Switching
Valve
INJ
Gradient:
3 mL/min
Symmetry C18 5 mm
3.9 mm x 150 mm
®
0.5 mL/min
Detector:
Injection Vol.
Total Run Time 5 min
Back-pressure:
MS/MS
Selected
Reaction
Monitoring
M. Jemal, M., et., Rapid Comm Mass Spec 1998, 12 pp 1389-1399
A switching valve is needed to divert the HPLC void volume to waste
in order to prevent proteins and other polar matrix interferences from
entering the MS source, thereby enhancing the MS performance.
0.0
© 2001 Waters Corporation
16
1.0
Minutes
Oasis® HLB cartridge column,
2.1 mm x 20 mm;
A = 100% 0.1% (aqueous)
formic acid, B = 95%
Acetonitrile:5% aqueous with 0.1%
formic acid in both
Flow Rate: 4 mL/min
0 to 0.2 min 100% A
0.2 to 0.8 min 100% B
Re-equilibrate 0.2 min at 100% A
Using splitter 0.4 mL/min to
mass spectrometer
50 µL of dilute plasma
(1:1 with mobile phase A)
Constant throughout the study
Sample Preparation
On-Line SPE With Oasis® HLB Sorbent
in Prospekt™ Cartridges
Sample Preparation
The Oasis® HLB sorbent in Prospekt™ cartridges shows better recoveries for
pharmaceutical compounds than silica-based sorbents, figure 21. Oasis®
HLB Prospekt™ cartridges give you rapid method development, simpler
methods, and equivalent or higher recoveries than previously achieved
even for the most challenging polar compounds.
Figure 21: Oasis® HLB Sorbent Superior to C18-Bonded Silica
Oasis HLB
®
C18
C2
0
Available Soon
™
Prospekt II
Cartridges
20
Clonidine
40
60
80
100
Terconazole
The general method in Figure 4 was used with 15% acetonitrile wash and 55%
acetonitrile for elution to the analytical column.
* Data were provided by Dr. Francis Beaudry, a Principal Research Scientist of Phoenix
International Life Sciences Inc. in Montreal, Quebec, Canada.
Oasis® Glass Cartridges for PPT Detection Levels
Waters Oasis® glass cartridges are available in 5 cc (200 mg) configuration with Teflon® frits for trace analysis at parts per trillion level. Each lot is
tested for the presence of bisphenol A and other phenols and phthalates,
assuring that endocrine disruptors in water samples can be analyzed to
part per trillion levels.
See the full application in the Oasis® Environ-mental & Agrochemical
Applications Notebook (Waters Literature Code #WN037).
PPT Recovery of Estrogens from River Water,
LC/MS, 5 ng/L Spike Level, n=4
Results
1. bisphenol A
2. 17β-estradiol
3. 17α-ethynylestradiol
4. estrone
5. diethylstilbestrol
Ions Monitored (m/z)
Recovery
% RSD
227
271
295
269
267
113
93
96
87
75
11
15
12
5
5
Recovery of Phthalates and Nonylphenol from
River Water, GC/MS, 200 ng/L Spike Level, n=4
Results
1. dimethylphthalate
2. dirthylphthalate
3. n-nonylphenol
4. dibutylphthalate
5. benzylbutylphthalate
6. bis(ethylhexyl)phthalate
ISTD. o-terphenyl (internal standard)
Recovery
% RSD
130
86
90
110
110
60
15
12
11
11
8
8
© 2001 Waters Corporation
17
Sample Preparation
Oasis® Cartridge Selection Guide
1 cc/10 mg
1 cc/30 mg
1 cc/30 mg
Gilson Adapter
3 cc/60 mg
3 cc/60 mg
Gilson Adapter
6 cc/150 mg
6 cc/200 mg
Box of 100
Box of 100
Box of 500
Box of 100
Box of 500
Box of 30
Box of 30
Oasis® HLB 30 µm
186000383
WAT094225
WAT058882
WAT094226
WAT058883
N/A
WAT106202
Oasis® HLB 60 µm
N/A
N/A
N/A
N/A
N/A
N/A
N/A
Oasis® MCX 30 µm
N/A
186000252
N/A
186000254
N/A
186000256
N/A
Oasis® MCX 60 µm
N/A
186000782
186000782
186000253
N/A
186000255
N/A
Oasis® MAX 30 µm
N/A
186000366
N/A
186000367
N/A
186000369
N/A
Oasis® MAX 60 µm
N/A
N/A
N/A
186000368
N/A
186000376
N/A
5 mg/96-well
10 mg/96-well
30 mg/96-well
60 mg/96-well
1/Pkg
1/Pkg
1/Pkg
1/Pkg
Oasis® HLB 30 µm
186000309
186000128
WAT058951
N/A
Oasis® HLB 60 µm
N/A
N/A
N/A
186000679
Oasis® MCX 30 µm
N/A
186000259
186000248
Oasis® MCX 60 µm
N/A
N/A
186000250
186000678
Oasis® MAX 30 µm
N/A
186000375
186000373
N/A
Oasis® MAX 60 µm
N/A
N/A
N/A
N/A
Sorbent
Oasis® 96-well Plates
© 2001 Waters Corporation
18
Sample Preparation
12 cc/500 mg
20 cc/1 g
35 cc/6 g
Plus
225 mg
Vac RC
30 mg
Vac RC
60 mg
Sample Preparation
6 cc/500 mg
Glass Cartridge
5 cc/200 mg
NEW
Box of 30
Box of 20
Box of 20
Box of 10
Box of 50
Box of 50
Box of 50
Box of 30
N/A
N/A
N/A
N/A
N/A
186000382
186000381
N/A
186000115
186000116
186000117
186000118
186000132
N/A
N/A
N/A
N/A
N/A
N/A
N/A
N/A
186000261
N/A
186000776
N/A
186000777
186000778
N/A
N/A
186000380
N/A
N/A
N/A
N/A
N/A
N/A
186000372
186000371
N/A
186000865
N/A
N/A
N/A
N/A
N/A
186000378
N/A
NEW 186000683
For the latest Oasis® applications visit
www.waters.com, choose HPLC Columns and
Supplies, then click on Sample Preparation.
Download the latest applications, or search our online Applications
Library containing thousands more applications covering the entire
range of Waters products.
If you do not have internet access, wish to receive printed copies, or
require further information, contact your local Waters Sales Office.
© 2001 Waters Corporation
19
Sample Preparation
Oasis® HLB Sample Extraction Products
Description
Particle Size
Quantity
Part Number
Price
Oasis HLB cartridge
Oasis® HLB cartridge
Oasis® HLB cartridge with Gilson ASPC™ Adapter
Oasis® HLB cartridge
Oasis® HLB cartridge with Gilson ASPC™ Adapter
Oasis® HLB cartridge
Oasis® HLB cartridge
Oasis® HLB cartridge
Oasis® HLB cartridge
Oasis® HLB cartridge
Oasis® HLB Plus cartridge
Oasis® HLB Vac RC cartridge
Oasis® HLB Vac RC cartridge
Oasis® HLB Glass cartridge
Adapter, 5 cc, Teflon®
Oasis® HLB Prospekt™ cartridge*
1 cc/10 mg
1 cc/30 mg
1 cc/30 mg
3 cc/60 mg
3 cc/60 mg
6 cc/200 mg
6 cc/500 mg
12 cc/500 mg
20 cc/1 g
35 cc/6 g
225 mg
30 mg
60 mg
5 cc/200 mg
5 cc
2 mm x 10 mm/15 mg
30 µm
30 µm
30 µm
30 µm
30 µm
30 µm
60 µm
60 µm
60 µm
60 µm
60 µm
30 µm
30 µm
60 µm
–
30 µm
100/box
100/box
500/box
100/box
500/box
30/box
30/box
20/box
20/box
10/box
50/box
50/box
50/box
30/box
10/box
100/box
186000383
WAT094225
WAT058882
WAT094226
WAT058883
WAT106202
186000115
186000116
186000117
186000118
186000132
186000382
186000381
186000683
405000934
186000258
???
???
???
???
???
???
???
???
???
???
???
???
???
???
???
???
Oasis® HLB column
Oasis® HLB cartridge column
Holder Kit for 2.1 mm x 20 mm cartridge column
In-Line pre-column filter kit
1 mm x 50 mm
2.1 mm x 20 mm
30 µm
25 µm
1/box
1/box
1/box
1/box
186000119
186000706
186000262
WAT084560
???
???
???
???
5 mg/96-well
10 mg/96-well
30 mg/96-well
60 mg/96-well
30 µm
30 µm
30 µm
60 µm
1/pkg
1/pkg
1/pkg
1/pkg
186000309
186000128
WAT058951
186000679
???
???
???
???
®
Oasis® HLB plate
Oasis® HLB plate
Oasis® HLB plate
Oasis® HLB plate
* For use with Spark Holland Prospekt™ system
Oasis® MCX Sample Extraction Products (Cation-Exchanger)
Description
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX cartridge
Oasis® MCX Vac RC cartridge
Oasis® MCX Vac RC cartridge
Oasis® MCX plate
Oasis® MCX plate
Oasis® MCX plate
Oasis® MCX Plate
Particle Size
Quantity
Part Number
Price
1 cc/30 mg
1 cc/30 mg LP**
3 cc/60 mg
3 cc/60 mg
6 cc/150 mg
6 cc/150 mg
6 cc/500 mg
20 cc/1 g
35 cc/6 g
6 cc/500 mg LP**
20 cc/1 g LP**
35 cc/6 g
60 mg
60 mg
30 µm
60 µm
30 µm
60 µm
30 µm
60 µm
60 µm
60 µm
60 µm
60 µm
60 µm
60 µm
30 µm
60 µm
100/box
100/box
100/box
100/box
30/box
30/box
30/box
20/box
10/box
30/box
20/box
10/box
50/box
50/box
186000252
186000782
186000254
186000253
186000256
186000255
186000776
186000777
186000778
186000776
186000777
186000778
186000261
186000380
???
???
???
???
???
???
???
???
???
???
???
???
???
???
10 mg/96-well
30 mg/96-well
30 mg/96-well
60 mg/96-well
30 µm
30 µm
60 µm
60 µm
1/pkg
1/pkg
1/pkg
1/pkg
186000259
186000248
186000250
186000678
???
???
???
???
** LP = Large Pore
© 2001 Waters Corporation
20
Sample Preparation
Oasis® MAX Sample Extraction Products (Anion-Exchanger)
Description
Particle Size
Quantity
Part Number
Price
1 cc/30 mg
3 cc/60 mg
3 cc/60 mg
6 cc/150 mg
6 cc/150 mg
6 cc/500 mg
30 mg
60 mg
60 mg
30 µm
30 µm
60 µm
30 µm
60 µm
60 µm
30 µm
30 µm
60 µm
100/box
100/box
100/box
30/box
30/box
30/box
50/box
50/box
50/box
186000366
186000367
186000368
186000369
186000370
186000865
186000372
186000371
186000378
???
???
???
???
???
???
???
???
???
Oasis® MAX plate
Oasis® MAX plate
10 mg/96-well
30 mg/96-well
30 µm
30 µm
1/pkg
1/pkg
186000375
186000373
???
???
Particle Size
Quantity
Part Number
Price
30 µm
10 each HLB, MCX, MAX
186000867
???
Sample Preparation
Oasis MAX cartridge
Oasis® MAX cartridge
Oasis® MAX cartridge
Oasis® MAX cartridge
Oasis® MAX cartridge
Oasis® MAX cartridge
Oasis® MAX Vac RC cartridge
Oasis® MAX Vac RC cartridge
Oasis® MAX Vac RC cartridge
®
NEW Oasis® Method Development Kit
Description
Oasis® Method Development Kit
3 cc/60 mg
Manifold for Extraction Plate
Description
Extraction Plate Manifold Kit A
(includes extraction plate manifold, reservoir tray,
manifold top gasket, sealing cap and 350 µL
sample collection plate)
Extraction Plate Manifold Kit B
(includes extraction plate manifold, reservoir tray,
manifold top gasket, sealing cap and 1 mL
sample collection plate)
Extraction Plate Manifold Kit C
(includes extraction plate manifold, reservoir tray,
manifold top gasket, sealing cap and
2 mL sample collection plate)
Extraction plate manifold
Accessories for Extraction Cartridges
Part No.
Price
WAT097944
???
WAT097945
???
Waters extraction manifold, 20-position without rack
(includes 20 needle tips, 25 plugs, and ejector tool)
Waters extraction manifold, 20-position
(complete with rack for 13 mm x 75 mm tubes)
Waters extraction manifold, 20-position
(complete with rack for 13 mm x 100 mm tubes)
Waters extraction manifold, 20-position
(complete with rack for 16 mm x 75 mm tubes)
Waters extraction manifold, 20-position
(complete with rack for 16 mm x 100 mm tubes)
Vacuum pump (110V, 60 Hz)
Vacuum pump (220V, 50 Hz)
Vacuum pump (110V, 50 Hz)
Reservoir 30 cc for Oasis cartridges
Reservoir 60 cc for Oasis® cartridges
Reservoir Adapter for 12, 20 and 35 cc cartridges
®
Quantity
Part No.
Price
48/box
12/box
10/box
WAT011390
WAT024659
WAT048160
???
???
???
Quantity
Part No.
Price
25/box
50/box
50/box
50/box
50 sheets/pkg
1/pkg
1/pkg
WAT058942
WAT058943
WAT058957
WAT058958
WAT058959
WAT058955
WAT058956
???
???
???
???
???
???
???
Accessories for Extraction Plate Manifold
WAT097946
???
WAT058941
???
Part No.
Price
WAT200677
???
WAT200606
???
WAT200607
???
WAT200608
???
WAT200609
WAT085114
WAT085115
WAT085123
???
???
???
???
Manifold for Extraction Cartridges*
Description
Description
Description
Reservoir tray
Sample collection plate, 350 µL
Sample collection plate, 1 mL
Sample collection plate, 2 mL
Sealing cap for 96-well collection plate
Manifold gasket, top
Manifold gasket, white
Waters Oasis® HLB Extraction Plate
30 mg/Well, 96-Wells/Plate
WAT058951
WAT058939
(1 Plate/Box)
(3 Plates/Box)
Sample Collection Plate
Polypropylene, 96-Wells,
50 Plates/Pkg
WAT058943
WAT058957
WAT058958
350 µL
1 mL
2 mL
Manifold Waste Tray
(Disposable)
WAT058942
25/Pkg
Top
Extraction Plate
Manifold
WAT058941
Bottom
© 2001 Waters Corporation
21
Sample Preparation
Sep-Pak® DNPH-Silica Cartridges for
Analyzing Formaldehyde, Other Aldehydes
and Ketones in Air
Formaldehyde and other aldehydes are receiving increasing attention
both as toxic substances and as promoters in the photochemical formation
of ozone in air. Source of aldehydes in residential buildings include plywood and particle board, insulation, combustion appliances, tobacco
smoke and various consumer products. Aldehydes are released into the
atmosphere in the exhaust of motor vehicles and other equipment in
which hydrocarbon fuels are incompletely burned.
Figure 6: HPLC Separation of DNPH Derivatives
of Common Aldehydes and Ketones
Column:
Sample:
Mobile Phase:
Gradient:
Flow Rate:
Injection:
Detection:
Nova-Pak® C18 3.9 mm x 150 mm
Mixture of DNPH and DNPH derivatives
in acetonitrile.
A: Water/Acetonitrile/Tetrahydrofuran
60/30/10 v/v/v
B: Water/Acetonitrile 40/60 v/v 3mL
100% A for 1 minute, then linear gradient from
100% A to 100% B in 10 minutes
1.5 mL/min
20 µL
360 nm
The most sensitive and specific method for analyzing aldehydes and
ketones is based on their reaction with 2,4-dinitrophenylhydrazine
(DNPH) and subsequent analysis of the hydrazone derivatives by HPLC
(see Figure 6). The hydrazones may be detected by absorbance in the
ultraviolet region, with maximum sensitivity obtained between 350 and
380 nm.
Airborne aldehydes have traditionally been collected by drawing a
sample through an impinger containing a solution of DNPH. However,
the impinger collector is generally cumbersome to use and is not well
suited for high flow rates or extended collection times due to solvent
evaporation.
The new Sep-Pak® DNPH-Silica cartridges meet the requirements of EPA
Method TO-11 and provide a convenient device for sample collection.
Using a vacuum pump, an air sample is drawn through the new SepPak® DNPH-Silica cartridge. The aldehydes and ketones react with the
DNPH and form the hydrazone derivative, which is retained on the cartridge. Later, the hydrazones are eluted from the cartridge with acetonitrile and analyzed by HPLC. Detection limits can be as low as 3 ppbv
for a 100 liter sample.
Advantages of Waters Sep-Pak®
DNPH-Silica Cartridges
These cartridges provide you with significant advantages when compared
to other techniques, such as liquid impingers, for the analysis of aldehydes
and ketones. In addition, a new high speed, high resolution HPLC
application has been developed to provide excellent quantitation
capability in the low parts-per-billion range (see Figures 7 and 8).
• Sep-Pak DNPH-Silica cartridges meet the requirements of
EPA Method TO-11 and ASTM-D-5791-1.
®
• Results from impingers and these cartridges are in excellent
agreement.
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
Acrolein-DNPH
Propionaldehyde-DNPH
Crotonaldehyde-DNPH
Butanone-DNPH
Butyraldehyde-DNPH
Benzaldehyde-DNPH
Isovaleraldehyde-DNPH
Valeraldehyde-DNPH
o-Tolualdehyde-DNPH
m-Tolualdehyde-DNPH
p-Tolualdehyde-DNPH
Hexaldehyde-DNPH
2,5-Dimethylbenzaldehyde-DNPH
Figure 7: Low-Level Example: Aldehyde Profile
from Laboratory Air
Lab Air Sample
1.
2.
3.
4.
5.
6.
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
Butanone-DNPH
Isovaleraldehyde-DNPH
Cartridge Blank (calculated for 100 liter sample)
1.
2.
3.
4.
4.8 ppbv
1.2 ppbv
118.0 ppbv
0.8 ppbv
0.7 ppbv
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
0.35 ppbv
0.27 ppbv
0.34 ppbv
The sample of Figure 7 was obtained in a chemical research laboratory using a portable sampling pump.
One hundred liters of air was drawn through the cartridge at 0.65 L/min. The sample was found to contain
low concentration of formaldehyde (4.8 ppbv), acetaldehyde (1.2 ppbv) and methyl ethyl ketone (0.8 ppbv),
but a significant amount of acetone (118 ppbv). Significant concentrations of acetone are frequently found in
laboratories because of its widespread use as a solvent.
Figure 8: High-Level Example: Aldehyde Profile
from Diluted Auto Exhaust Emissions
Sample (8.4 L at 1 L/min) collected at US EPA
Atmospheric Research Assessment Lab. Courtesy
of Dr. S. B. Tejada.
• Solvent consumption, solvent exposure and hazardous waste
disposal costs are reduced.
• Sep-Pak® DNPH-Silica cartridges provide superior convenience
and reproducibility, making them ideal for field sampling and
process monitoring applications.
• Sep-Pak® DNPH-Silica cartridges can save time and increase
productivity.
• Increased safety.
© 2001 Waters Corporation
22
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
DNPH
Formaldehyde-DNPH
Acetaldehyde-DNPH
Acetone-DNPH
Acrolein-DNPH
Propionaldehyde-DNPH
Crotonaldehyde-DNPH
2-Butanone-DNPH
Butyraldehyde-DNPH
Benzaldehyde-DNPH
o-Tolualdehyde-DNPH
m-p-Tolualdehyde-DNPH
2,5-Dimethylbenzaldehyde-DNPH
2290 ppbv
367 ppbv
26 ppbv
8 ppbv
12 ppbv
36 ppbv
24 ppbv
15 ppbv
32 ppbv
5 ppbv
11 ppbv
3 ppbv
Sample Preparation
Waters XPoSure™ Aldehyde Sampler Cartridges
for Monitoring Aldehydes in indoor air.
Sample Preparation
Based on an extension of our DNPH coating technology, XPoSure™
Aldehyde Sampler cartridges are the most sensitive active samplers
available today.
Figure 9: Low-Level Example: Aldehyde
Profile from Laboratory Air
Highest Sensitivity
4
1
Compared to existing sampling tube technology which have high
and variable backgrounds, XPoSure™ cartridges are guaranteed to
give consistent low aldehyde backgrounds, cartridge-to-cartridge,
lot-to-lot.
1.
2.
3.
4.
5.
6.
DNPH
Formaldehyde-DNPH
Acetalhyde-DNPH
Acetone-DNPH
Butanone-DNPH
Isovaleraldehyde-DNPH
High-Collection Efficiencies
You can achieve > 95% collection efficiencies for all aldehydes at
flows of up to a liter per minute. And, you only need to use one
cartridge - no breakthrough bed is necessary.
2
3
Low Pressure Drop - Use with Portable Personal
Sampling Pumps
5
6
Lab Air Sample
Large particle size and higher porosity frits make this new XPoSure
cartridge compatible with personal sampling pumps.
™
Easy-to-Use
Cartridge Blank
0
Sample, elute and shoot, it’s that easy. You’ll never have to break
open and manipulate a glass tube again. And because the cartridges
are made from high density polyethylene (HDPE), breakage is not
a concern.
2
4
6
8
Time (minutes)
10
12
14
The above sample was collected in a chemical research laboratory using a portable sampling pump.
One hundred liters of air was drawn through the cartridge at 0.65 L/min. The chromatogram shows
ormaldehyde (4.8 ppbv), acetaldehyde (1.2 ppbv), acetone (118 ppbv), butanone (0.8 ppbv), and
isovaleraldehyde (0.7 ppbv).
The figure below shows two traces. An actual cartridge blank
demonstrating extremely low background levels and as an actual
laboratory air sample.
Ozone Scrubber Cartridge
Ozone has been shown to interfere with the analysis of carbonyl
compounds in air samples that have been drawn through cartridges
containing silica-coated with
Flow Schematic for Air Sampling System
2,4-dinitrophenylhydrazine (DNPH). Waters Ozone Scrubber cartridges
are designed to remove this ozone interference.
These disposable devices are intended for use in series combination with
the Waters Sep-Pak® DNPH-Silica cartridges or XPoSure™ Aldehyde
Sampler cartridges. One Ozone Scrubber cartridge replaces the 1⁄4”
diameter by 36” long copper ozone denuder located in the heated zone
of sampling systems used for outdoor air monitoring (PAMS program).
Air
Sample
In
O3 Scrubber
Cartridge
DNPH-Silica
or XPoSure™
Cartridge
Vacuum
Sampling
Pump
Each Ozone Scrubber cartridge contains 1.4 grams of granular potassium iodide. When air containing ozone is drawn through this packed
bed, iodide is oxidized to iodine, consuming the ozone, according to the
following reaction:
The theoretical capacity of a single cartridge is 4.2 mmoles of ozone
(200 mg).
The particle size of the potassium iodide granules is optimized for good
mass transfer and flow characteristics.
© 2001 Waters Corporation
23
Sample Preparation
Porapak® RDX Sep-Pak® Extraction Cartridge
for the Analysis of Explosives in Surface and
Ground Waters.
Designed to meet or exceed the QA/QC requirements of EPA method
8330, it is ideal for environmental testing laboratories supporting
Department of Defense remediation programs.
High Sensitivity
Porapak® Sep-Pak® cartridges contain Porapak® RDX resin, a specially
prepared, specially cleaned divinylbenzene/vinylpyrrolidone copolymer,
packed in a high purity polyethylene syringe barrel. With the lowest
guaranteed backgrounds and the highest cartridge-to-cartridge, lot-to-lot
consistency, The Waters Porapak® RDX column is the most sensitive
technology available today and allows you to perform analysis at
sub ppb levels.
Increase Productivity and Reduce waste
By using Porapak® RDX cartridges, you can reduce the amount of organic
solvent used per sample by 10-fold and decrease your sample prep
time by 3x.
Activate, Load, Elute and Shoot
Flow 15 mL acetonitrile through
cartridge at 10 mL/min
Flow 30 mL water through
cartridge at 10 mL/min
Load 500 mL water sample
at 10 mL/min
Elute with 5 mL acetonitrile
Unmatched Recoveries
The specially prepared resin is highly selective for nitroaromatic and
nitramine compounds, resulting in recoveries of 90% or greater.
Dilute to 40% acetonitrile
with HPLC grade water
Inject 100 µL onto HPLC
Recovery data from preconcentrating 500 mL of explosives standards
in sterile water at two concentrations on Porapak® RDX Sep-Pak® Vac
columns. Number of replicates = 7.
1 ppb
% Recovery
% RSD
10 ppb
% Recovery
% RSD
HMX
TNB
RDX
100.5
95.9
90.9
6.7
3.5
6.4
100.5
99.3
98.7
3.9
3.3
3.2
DNB
TNT
TETRYL
99.5
97.0
89.0
3.2
3.0
6.4
99.2
102.
102.8
3.2
3.7
4.7
NB
35DNA
24DNT
96.5
91.2
97.3
2.5
3.3
3.4
97.9
98.2
99.9
2.8
3.6
3.4
26DNT
2A
4A
94.5
92.4
90.0
3.4
5.2
4.9
98.7
98.0
97.2
3.4
3.7
4.1
4NT
2NT
89.5
96.8
4.3
6.6
100.4
93.4
3.7
3.0
Compound
Figure 10: Isocratic Separation of Method 8330 Analytes
Column:
Mobile Phase:
Flow rate:
Injection volume:
Temperature:
Detection:
Sample:
Waters Nova-Pak® C8, 3.9 x 150 mm
82:18 water/isopropanol
1.0 mL/min
100 mL
24°C
UV @ 254 nm
1 ppb explosives concentrated
40-fold on Porapak® RDX, in 60:40
water/acetonitrile
Achieve the lowest method detection limits using Waters Porapak® RDX
cartridges. The top chromatogram shows the analysis of reagent H2O
spiked at the 1 ppb level. The bottom chromatogram is a reagent
(H2O) blank.
© 2001 Waters Corporation
24
Sample Preparation
Sep-Pak® DNPH-Silica Cartridge
XPosure™ Aldehyde Sampler Cartridge
Quantity
Part No.
Price
Sep-Pak® DNPH-Silica Cartridge
Box of 20
WAT037500
195.00
Ozone Scrubber
Description
Description
Quantity
Part No.
Price
XPoSure™ Aldehyde Sampler Cartridge
Box of 20
WATO47205
195.00
Porapak® RDX Cartridges and Accessories
Quantity
Part No.
Price
Description
Quantity
Porapak RDX Cartridges
Box of 30
Tubing, Tefzel, 1/8 inch O.D. x 0.040-inch I.D.
Sep-Pak® Vac Adapter
Box of 12
60 cc Sep-Pak Reservoir
Box of 12
Male-Male Adapter
®
Ozone Scrubber
Sample Preparation
Description
Box of 20
WATO54420
105.00
Part No.
Price
WAT047220
WAT023344
WAT054260
WAT024659
WAT024310
180.00
68.00
35.00
12.00
55.00
Sep-Pak® C18 Environmental Cartridges
Description
Quantity
Part No.
Price
C18 Environmental Cartridge
tC18 Environmental Cartridge
Box of 50
Box of 50
WAT023635
WAT036800
168.00
168.00
TM
Visit Waters on-line
product resource today
© 2001 Waters Corporation
25
Sample Preparation
Waters Solid-Phase Extraction Applications Guide
and Bibliography, 6th Edition. A resource for
Sample Preparation Methods Development.
Designed to be easy-to-use, the 600 page bibliography has
separate indexes by:
Compound
Now make your methods development effort easier with references to
over 3,000 published papers utilizing Sep-Pak® cartridges in sample
preparation protocols. The 6th Edition bibliography has more than 600
pages, contains 3,000 plus referenced papers, more than 8,000 compound entries, 8,000 author entries and more than 1,000 sample matrix
entries. It also has an expanded sample preparation primer to assist in
your development of a sample preparation protocol.
Sample
Matrix
Author
To help you in your methods development, see information on Sep-Pak®
Plus Methods Development Kits below.
Paper reference file number (numerical sequence)
Publication (page reference)
Application category
Sample matrix
Title
Cartridge selection
Compound(s)
Analytical technique
Author(s)
Now Online - Click Here to Search
Waters Solid-Phase Extraction Applications
Guide and Bibliography, 6th Edition.
Solid-Phase Extraction Technology Training Video
Description
Part No.
Price
Waters Solid-Phase Extraction Applications Guide
and Bibliography, 6th Edition
WAT052853
75.00
Experiments in Chromatography: Solid-Phase Extraction Technology, an
18 minute video designed for use in the training of laboratory personnel
and chemistry students.
Description
Experiments in Chromatography
Experiments in Chromatography
Sep-Pak® Plus Methods Development Kits
Description
Quantity
Methods Development Kit A
Box 100
(contains 10 Sep-Pak® cartridges each of Alumina:
Acid, Basic, Neutral, CN, Diol, Silica, Florisil,
Accell Plus QMA, C18, and NH2)
Methods Development Kit B
Box 50
(contains 10 Sep-Pak® cartridges each of NH2, CN,
Diol, Silica and C18)
Methods Development Kit C
Box 50
(contains 10 Sep-Pak® cartridges each of Accell Plus QMA,
Accell Plus CM, C18, Silica and Diol)
Part No.
Price
WAT039545
53.00
WAT039555
53.00
WAT039565
53.00
Format
Part Number
Price
NTSC
PAL
WAT054295
WAT054325
16.00
16.00
Sep-Pak® Cartridge Experiment Kit
This activity is designed to demonstrate the principles of liquid chromatography using a grape drink mix and Sep-Pak® C18 Plus cartridges. This
experiment lends itself ideally to a classroom setting.
Description
Cartridge Experiment Kit (Includes two Sep-Pak C18 cartridges
and 1 pack of grape mix with instructions)
Part No.
Price
WAT088253
15.00
®
© 2001 Waters Corporation
26
Sample Preparation
Solid-Phase Extraction
Sep-Pak® cartridges are available in a variety of designs; each offers
specific functional benefits but all contain the same high-quality packing
sorbents. This makes transfer of methods from one design to another
straightforward and predictable.
Sep-Pak® Plus Cartridges for Maximum Versatility
Sep-Pak® Plus cartridges are designed for both convenient manual use
and compatibility with automated instruments. They come in short body
and long body configurations and are constructed from high purity
polyethylene with female Luer inlets and male Luer outlets. They can be
connected to many different types of devices such as positive flow pumps
and syringes, vacuum manifolds, and automated sample processors.
In many applications, the use of positive flow (pumps) provides for
increased reproducibility in your results. Variations in recovery data, due
to flow rate variability and the potential for air drying out a cartridge
when using vacuum devices, can be eliminated.
The male outlet tips feature reduced hold-up volume for precise fraction
collection. The fittings also allow cartridges to be easily stacked for
increased capacity or for the use of more than one sorbent type for
cleanup of complex sample mixtures.
Sep-Pak® Plus cartridges can be used with liquid or gas samples and
have the unique capability for reverse flow direction, a useful feature for
trace enrichment applications. The packing process and compression seal
provide a uniform bed structure to minimize band spreading for the best
possible separation efficiency.
Bed volumes and weights of sorbent in the short and long body
cartridges equal those of the corresponding Classic cartridges.
All Sep-Pak® Plus cartridges have color-coded compression rings for
general sorbent identification.
Sep-Pak® Classic Cartridges
Sep-Pak® Classic cartridges were the original SPE cartridges introduced
over 20 years ago. Waters Sep-Pak® Classic cartridges contain the same
amount of sorbent as the Sep-Pak® Plus cartridge. This means that you can
convert existing methods easily to the more convenient Plus configuration.
Sep-Pak® Light Cartridges for Limited Sample
Volumes/Lower Solvent Consumption
Sep-Pak® Light cartridges look similar to the short-body Sep-Pak® Plus
cartridges, except for the distinctive “finned” outer body. The functional
difference is the reduced internal diameter which results in bed volumes
about one-third that of the corresponding Plus cartridge. This design
allows you to elute fractions in a minimal volume to improve recoveries
and reduce solvent consumption, a benefit for “sample-limited” applications or where excessive dilution is a concern. In addition, the Sep-Pak®
Light cartridges are specially designed to provide efficient packed sorbent
beds for use in downsizing current applications to reduce solvent
consumption. All Sep-Pak® Light cartridges have color-coded compression
rings for general sorbent identification.
Sep-Pak® Vac RC Cartridges for Manual
or Robotic Sample Preparation
These cartridges feature a 20 cc “funnel” shape design containing 100
or 500 mg of sorbent. They can be used with robotic sample preparation equipment or with vacuum manifolds. The integral reservoir design
allows for the convenient processing of larger amounts of sample or
solvents. This reduces setup time and costs associated with the use of
separate reservoirs.
Color-coded printing on the cartridges makes sorbent identification easy.
Economical Sep-Pak® Vac Cartridges
Sep-Pak® Vac cartridges provide you with the option of even lower cost
sample preparation. The cartridges use molded polypropylene “syringebarrel” bodies and two polyethylene filters to contain the packing
material. A standard male Luer outlet tip permits use with most multiposition vacuum manifolds. Cartridge sizes range from 1 cc to 35 cc
and sorbent weights range from 50 mg up to 10 grams. Sizes up to
6 cc can be used on many automated sample preparation systems
designed to use “syringe-barrel” style cartridges. Sep-Pak® Vac cartridges
offer the highest quality at low cost with a complete range of capacities
for use in optimizing your methods.
Color-coded printing on the syringe-barrel cartridges makes sorbent
identification easy.
Both the Sep-Pak® Plus and Classic designs have two different sizes (short
body and long body) depending on the type of sorbent.
Sep-Pak® C18 Environmental Cartridges
This is a special “long body” Plus cartridge design containing
approximately 1 gram of the monofunctional C18 or the trifunctional
C18 sorbent. These cartridges are especially useful in trace
enrichment applications.
© 2001 Waters Corporation
27
Sample Preparation
Which Cartridge Design is Best for Your Application
Sample Preparation
The Anatomy of Sep-Pak® Plus and
Classic Cartridge Design
The Anatomy of Sep-Pak® Vac and
Vac RC Cartridge Designs
Sep-Pak® Classic
Sep-Pak® Plus
Sep-Pak® Vac
Sep-Pak® Vac RC
1
7
5
2
3
4
5
6
6*
1
7
2
3
11
10
4
8
8
9
6
9
1.
Female Luer inlet accepts male Luer tip. Plus cartridge design can be stacked.
2.
Highest quality sorbents designed and made specifically for sample
preparation; clean, dry, reproducible in activity and capacity with optimal
surface area, pore, and particle size distributions.
2.
Broad range of sorbent surface activities available; each lot is tested under
rigid specifications for chromatographic activity, retention and selectivity.
Broad range of sorbent surface activities available; each lot is tested under
rigid specifications for chromatographic activity, retention and selectivity.
3.
Sorbent type and bed dimensions equal to corresponding Sep-Pak® Classic
cartridges enable direct transfer of previously developed and published
methods to new Plus design.
Weight of sorbent in each cartridge is controlled within +/- 5% of
specification to assure reproducible performance.
4.
Special blend of HD and UHMW polyethylenes used for 20 micron frits.
5.
Molded, medical grade polypropylene body.
5.
Weight of sorbent in each cartridge is controlled within +/- 5% of
specification to assure reproducible performance.
6.
Cartridges are sealed in a special polyfoil pouch to protect product integrity,
sorbent activity and purity.
6.
Advanced bed formation technology to minimize voids and channels. 6*
Patented Radial Compression Technology used to form homogeneous packed
bed free of voids and channels.
7.
Integral reservoir approximately 20 mL, robotic compatible.
8.
Outlet male Luer tip.
9.
Color-coded labeling on the cartridge to identify the sorbent.
3.
4.
7.
Special blend of HD and UHMW polyethylenes used for 20 micron frits
imparts excellent solvent resistance, extremely low extractables level, and
good flow properties. Frit also acts as depth filter for small amounts of
sample debris.
8.
Polyethylene body has excellent solvent resistance. All body parts are quality
tested to verify extremely low level of UV-absorbing extractables. Plus design is
molded for precise dimensions making it suitable for automated equipment.
9.
Cartridges are sealed in a special polyfoil pouch to protect product integrity,
sorbent activity and purity.
1. Highest quality sorbents designed and made specially for sample
preparation; clean, dry, reproducible in activity and capacity, with
optimal surface area pore and particle size distributions.
10. Male Luer outlet has reduced internal volume for minimal sample hold-up.
11. Color-coded ring compresses and seals the cartridge and identifies sorbent.
P.D. McDonald, C. W. Rausch, Radial Compression of Packed Beds, U.S. Patent # 4,250,035 (1981); Great Britain # 1,569,700 (1976); Canada # 1,101,785 (1981);
Japan # 1,400,983 (1987); Sweden # 450,750 & # 453,437 (1987); Germany # 2,655,650 (1988); other patents pending.
© 2001 Waters Corporation
28
Sample Preparation
Which Sorbent is Best For Your Application
The text and tables that follow give you detailed information on the
properties of the sorbents that Waters offers.
The Original C18 Sorbent
Waters original C18 sorbent is the most widely referenced SPE product,
with over 2,300 published papers or articles specifying its use. This
product is made using the most reproducible bonding technology
available, utilizing monofunctional silane chemistry. Waters original C18
sorbent offers a unique selectivity and excellent recovery of analytes, and
is recommended for most applications where HPLC is utilized for
subsequent analysis.
tC18 (trifunctional) Sorbent
With the addition of tC18 sorbent, you can now choose between two
highly reproducible C18 chemistries, monofunctional and trifunctional.
Trifunctional tC18 cartridges are recommended for most applications
involving GC, GC/MS or LC/MS especially if long sample contact
times are involved. This new tC18 sorbent has also become increasingly
popular in pharmaceutical applications when a high carbon load, very
hydrophobic surface is required. (See Table 1).
Table 1: Recommendations for Using
C18 and tC18 Sorbents
Sample Contact Time
long
(> 10 minutes)
short
(< 10 minutes)
Sample pH
<3
3-7
>7
Detection Method
HPLC
Ultraviolet/Visible Absorbance
Fluorescence
GC
Flame Ionization
Mass Selective
Electron Capture
Nitrogen-Phosphorous
Selectivity
Unique
Similar to other vendors’ C18 cartridges
Key:
R = recommended sorbent
O = may also be applicable
C18
tC18
NR
R
R
O
NR
R
NR
R
O
R
R
R
O
O
O
O
O
O
R
R
R
R
R
NR
NR
R
Sample Preparation
Selecting the most appropriate sorbent is best approached from both a
theoretical and empirical standpoint. Knowledge of the components in a
specific sample will help you determine the general mode of separation,
such as, normal phase, reversed-phase, or ion exchange, and more
specific knowledge may help you select between the various types of
sorbents in each mode. However, it is frequently necessary to determine
the optimal separation by empirical means.
NR = not recommended
tC2 (trifunctional) Sorbent
Many applications require a less hydrophobic (relative to C18), short
carbon chain sorbent to provide optimal sample processing of a wide
range of analytes. However, finding a reproducible, stable C2 sorbent
can be difficult. Waters has recently developed a new, state-of-the-art
trifunctional bonding process that provides you with the most dependable
sorbents (tC2 and tC18) of this type available. The tC2 is ideal for many
types of applications such as environmental and pharmaceutical.
TM
Visit Waters on-line
product resource today
© 2001 Waters Corporation
29
Sample Preparation
Table 2: Sorbent Properties and Typical Applications
Separation Mode
Sorbent Properties & Applications
Reversed-Phase
C18 silica-based bonded phase with strong hydrophobicity; used to adsorb analytes of even weak hydrophobicity from aqueous solutions; typical applications include drugs and their
metabolites in serum, plasma or urine, desalting of peptides, trace organics in environmental water samples, organic acids in beverages; similar in behavior to reversed-phase
HPLC columns
tC18 silica-based bonded phase with strong hydrophobicity; trifunctional bonding chemistry gives it an increased hydrolytic stability over C18; applications similar to C18
C8 silica-based bonded phase with moderate hydrophobicity; use for methods requiring less retention than C18; typical applications include drugs and their metabolites in serum,
plasma or urine, peptides in serum, plasma
tC2 silica-based bonded phase with low hydrophobic character; use for methods requiring less retention than C8; applications are similar to C18 and C8
Porapak RDX specially cleaned polymeric poly(divinylbenzene-vinylpyrrolidone) resin with hydrophobic character. Can be used as an alternative to octadecyl-bonded silica for preparation of
analytes that weakly adsorb to silica-based reversed-phase sorbents. Compatible with sample or eluents at high and low pH. Specifically designed for the concentration of
high explosives in aqueous samples.
™
Reversed or Normal Phase:
Amino Propyl (NH2) silica-based polar bonded phase with basic character; can be used as a polar sorbent, like silica, with different selectivity for acidic/basic analytes or as weak anion exchanger
in aqueous medium; applications include phenols and phenolic pigments, petroleum fractionation, saccharides and drugs and metabolites
Cyano Propyl (CN) silica-based polar bonded phase; can be used as less polar alternative to silica in normal-phase applications or as less hydrophobic alternative to C18 or C8 in reversed-phase
applications; typical applications include drugs, drug metabolites and pesticides
Diol silica-based polar bonded phase with neutral character; can be used as an alternative to silica in normal phase applications, where the acidic character of silica is undesirable or
as very weakly interacting phase in aqueous applications; applications include antibiotics from cosmetics; isolation of proteins or peptides by hydrophobic interaction chromatography
Normal Phase
Silica polar sorbent, used primarily to adsorb analytes from non polar solvents like hydrocarbons, chloro- or fluoro-substituted hydrocarbons or less polar esters and ethers; elution with
more polar solvents like polar esters, ethers, alcohols, acetonitrile or water; the binding mechanism can be hydrogen bonding or dipole-dipole interaction; silica can also be used in
aqueous medium as a cation exchanger of intermediate strength.
Alumina (A, B & N) similar in use to silica; available in acidic, basic and neutral high activity grades; alumina also exhibits specific interactions with the pi-electrons of aromatic hydrocarbons, making
it useful for applications like crude oil fractionation; acidic and basic grades can also be used as low-capacity ion-exchangers
Florisil® polar, highly active, weakly basic sorbent for adsorption of low to moderate polarity species from nonaqueous solutions; specifically designed for the adsorption of pesticides using
official AOAC and EPA methods; other applications include polychlorinated biphenyls (PCB’s) in transformer oil
Ion-Exchange
Accell™ Plus QMA silica-based hydrophilic strong anion-exchanger with large pore-size; extraction of anionic analytes in aqueous and non-aqueous solutions; due to the large pore-size, it is excellent
for the isolation of anionic proteins, e.g., immunoglobulins, enzymes; other applications include the removal of acidic pigments from wines, fruit juices and food extracts, isolation
of phenolic compounds, peptide pool fractionations
Accell™ Plus CM silica-based hydrophilic weak cation-exchanger with large pore-size; extraction of cationic analytes in aqueous and non-aqueous solutions; due to the large pore-size, it is excellent
for the isolation of cationic proteins; other applications include pesticides, herbicides, steroids
Specialty
DNPH-Silica, XPoSure™ contains acidified dinitrophenylhydrazine reagent coated on a silica sorbent. Used for the collection of air samples and subsequent quantitation of aldehydes and ketones by
reaction to form the hydrazone derivative, and analysis by HPLC. DNPH-Silica is specified in several EPA procedures for the analysis of carbonyl compounds in air.
© 2001 Waters Corporation
30
Sample Preparation
Table 3: Sep-Pak® Cartridge Sorbents Physicochemical Characteristics
Endcapped
Percent
Carbon
Pore Size
(nominal, Å)
Particle Size
Range (µm)
C18
tC18
N/A
N/A
Yes
Yes
12
17
125
125
55-105
37-55
C8
N/A
Yes
9
125
37-55
tC2
N/A
Yes
2.7
125
37-55
NH2
N/A
No
3.5
125
55-105
CN
N/A
Yes
6.5
125
55-105
Diol
N/A
No
2
300
37-55
Silica
High*
N/A
N/A
125
55-105
Florisil
High**
N/A
N/A
60
50-200
Alumina A
1***
N/A
N/A
120
50-300
Alumina N
1***
N/A
N/A
120
50-300
Alumina B
1***
N/A
N/A
120
50-300
Accell Plus CM
N/A
N/A
5.5
300
37-55
Accell Plus QMA
N/A
N/A
6
300
37-55
Porapak RDX
N/A
N/A
95
200
125-150
DNPH-Silica
N/A
N/A
N/A
125
55-105
XPoSure
N/A
N/A
N/A
125
500-1000
***
Moisture content ≤ 1.5%; activity ≤ 1 on Brockmann activity scale
®
™
™
®
™
* Moisture content ≤ 3.2%
** Moisture content ≤ 2.5%
Sample Preparation
Activity
Grade
Sorbent
Table 4: Sep-Pak® Cartridges: Chemistry of Bonded-Phase Sorbents
Sorbent
Chemistry
Bonding Chemistry
Surface Functionality
C18
Monofunctional silane
–Si(CH3)2C18H37
tC18
Trifunctional silane
–SiC18H37
C8
Monofunctional silane
–Si(CH3)2C8H17
tC2
Trifunctional silane
-SiC2H5
NH2
Trifunctional silane
–Si(CH2)3NH2
CN
Difunctional silane
–Si(CH3)(CH2)3CN
Diol
Trifunctional silane
–Si(CH2)3OCH2CH(OH)CH2OH
Accell Plus CM
Polymeric*
–CO2- Na+
™
Accell Plus QMA
Polymeric**
–C(O)NH(CH2)3N(CH3)3+ Cl-
DNPH-Silica
Coated***
™
* An acrylic acid/acrylamide copolymer on Diol silica; ligand
density: 350 µmoles/g; approximate protein binding
capacity = 175 mg Cytochrome c/g of sorbent or
63 mg/Accell Plus CM Classic or Plus cartridge.
** An acrylamide copolymer on Diol silica; ligand density:
220 µmoles/g; approximate protein binding
capacity = 200 mg BSA/g of sorbent or 72 mg/
Accell Plus QMA Classic or Plus cartridge.
*** Unbonded silica sorbent coated with 14 µmoles DNPH/g
of sorbent or 5 µmoles/cartridge.
© 2001 Waters Corporation
31
Sample Preparation
Table 5: Nominal Weight of Sorbent (in milligrams)
Separation Mode
and Sorbent
Cartridge Type
Classic
Light
General Surface
Chemistry
360
130
Neutral
N/A
145
Neutral
N/A
N/A
145
145
Neutral
Neutral
360
360
360
360
360
360
130
130
130
Basic
Neutral
Neutral
690
910
1710
1710
1710
690
900
1850
1850
1850
120
145
280
280
280
Neutral
Basic
Acidic
Basic
Neutral
360
360
360
360
130
130
Basic
Acidic
Plus
Reversed-phase
360
820
400
900
400
400
C18
Environmental C18*
tC18
Environmental tC18*
C8
tC2
Normal or Reversed-phase
Amino Propyl (NH2)**
Cyano Propyl (CN)
Diol
Normal Phase
Silica
Florisil®
Alumina A
Alumina B
Alumina N
Ion-Exchange **
Accell™ Plus QMA
Accell™ Plus CM
*
**
Sep-Pak® Plus long-body cartridge.
The amino propyl packing may also be used in the protonated form as a weak anion exchanger.
Table 6: Sep-Pak® Vac Cartridges Nominal Weight of Sorbent
Syringe Size
1 cc
50 mg
100 mg
X
X
Vac RC
3 cc
200 mg
500 mg
X
X
X
6 cc
X
20 cc
2g
5g
X
X
X
35 cc
X
See Cartridge Selection Guide for specific product availability.
© 2001 Waters Corporation
32
10 g
X
Vac RC
12 cc
1g
Sample Preparation
Normal Phase Chromatography
with Sep-Pak® Cartridges
Sample Preparation
To perform normal phase chromatography with Sep-Pak®
cartridges, use a gradient of nonpolar solvents with polar
Silica, Florisil®, NH2, Diol, CN, Alumina A, B, or N
as a sorbent*.
1. You may condition the cartridge with six to ten hold-up
volumes of nonpolar solvent, usually the sample solvent.
2. Load the sample into the cartridge.
3. Elute unwanted components with a nonpolar solvent.
4. Elute the first component of interest with a polar solvent.
5. Elute remaining components of interest with progressively
more polar solvents.
6. When you recover all of your components, discard the used
cartridge in an appropriate manner.
* Depending upon your chromatographic conditions, you
may also use CN as a packing material for normal phase
chromatography.
Reversed-Phase Chromatography
with Sep-Pak® Cartridges
To perform reversed-phased chromatography with Sep-Pak®
cartridges, use a gradient of strongly to weakly polar solvents with
nonpolar C18, tC18, C8, tC2, Diol, NH2 or CN as a sorbent.
1. Solvate the bonded phase with six to ten cartridge hold-up
volumes of methanol or acetonitrile. Flush the cartridge with
six to ten hold-up volumes of water or buffer. Do not allow
the cartridge to dry out.
2. Load the sample dissolved in strongly polar solvent.
3. Elute unwanted components with a strongly
polar solvent.
4. Elute weakly held components of interest with a less
polar solvent.
5. Elute more tightly bound components with progressively more
nonpolar solvents.
6. When you recover all of your components, discard the used
cartridge in an appropriate manner.
Ion-Exchange Chromatography
with Sep-Pak® Cartridges
To perform ion-exchange chromatography with Sep-Pak®
cartridges, use a gradient of pH or ionic strength with
Accell™ Plus CM, Accell Plus QMA or NH2 as a sorbent.
1. Condition the cartridge with six to ten hold-up
volumes of deionized water or weak buffer.
2. Load the sample dissolved in a solution of deionized water
or buffer.
3. Elute unwanted weakly bound components with a
weak buffer.
4. Elute the first component of interest with a stronger buffer
(change the pH or ionic strength).
5. Elute other components of interest with progressively
stronger buffers.
6. When you recover all of your components, discard the used
cartridge in an appropriate manner.
© 2001 Waters Corporation
33
Sample Preparation
Sep-Pak® Cartridge Selection Guide
Vac 3 cc
200 mg
Waters Sep-Pak® Vac
Classic
Light
Vac 1 cc
50 mg
Box of 50
Box of 50
Box of 50
C18
WAT020515
WAT051910
WAT023501
tC18
WAT036810
N/A
C8
WAT036775
tC2
Vac 1 cc
100 mg
Box of 100
Box of 100
Box of 50
Box of 50
WAT054955
WAT023590
WAT036935
WAT054945
WAT036805
WAT054960
WAT036820
WAT043410
WAT054925
N/A
WAT036770
WAT054965
WAT036785
WAT043415
WAT054940
WAT052720
N/A
WAT052725
N/A
WAT052710
N/A
N/A
Silica
WAT020520
WAT051900
WAT023537
WAT054980
WAT023595
WAT036940
WAT054930
Florisil®
WAT020525
WAT051960
WAT023543
WAT054985
WAT023600
WAT043420
N/A
Accell™ Plus CM
WAT020550
WAT010910
WAT023531
N/A
WAT023625
WAT043465
N/A
Accell™ Plus QMA
WAT020545
WAT010835
WAT023525
N/A
WAT023620
WAT043460
N/A
Alumina A
WAT020500
WAT051800
WAT023549
N/A
WAT023575
WAT043450
N/A
Alumina B
WAT020505
WAT051820
WAT023555
N/A
WAT023580
WAT043455
N/A
Alumina N
WAT020510
WAT051810
WAT023561
N/A
WAT023585
WAT043445
N/A
Amino Propyl (NH2)
WAT020535
WAT010830
WAT023513
N/A
WAT023610
WAT043475
N/A
Cyano Propyl (CN)
WAT020540
WAT010823
WAT023507
WAT054975
WAT023615
WAT043470
WAT054935
Diol
WAT020530
WAT010818
WAT023519
N/A
WAT023605
WAT043480
N/A
Waters Sep-Pak® Vac
Vac RC
100 mg
Waters Sep-Pak® Vac
Plus
Sorbent
mg = nominal weight of sorbent in milligrams.
See Sep-Pak® cartridge configuration and nominal weight of sorbent chart on
page 32 for specific values of each sorbent.
© 2001 Waters Corporation
34
Sample Preparation
Vac RC
500 mg
Vac 6 cc
500 mg
Vac 6 cc
1 gram
Vac 12 cc
2 gram
Vac 20 cc
5 gram
Vac 35 cc
10 gram
Waters Sep-Pak® Vac
Waters Sep-Pak® Vac
Waters Sep-Pak® Vac
Waters Sep-Pak® Vac
Waters Sep-Pak® Vac
Waters Sep-Pak® Vac
Sample Preparation
Vac 3 cc
500 mg
Sorbent
Box of 50
Box of 50
Box of 30
Box of 30
Box of 20
Box of 20
Box of 10
WAT020805
WAT036945
WAT043395
WAT036905
WAT036915
WAT036925
WAT043345
C18
WAT036815
WAT043425
WAT036790
WAT036795
WAT043380
WAT043365
WAT043350
tC18
WAT036780
WAT043430
WAT054525
WAT054570
WAT054615
WAT054660
WAT054700
C8
WAT052715
N/A
N/A
WAT052705
N/A
N/A
N/A
tC2
WAT020810
WAT036950
WAT043400
WAT036910
WAT036920
WAT036930
WAT043355
Silica
WAT020815
WAT043435
WAT043405
WAT043390
WAT043385
WAT043370
WAT043360
Florisil®
WAT020855
WAT054505
WAT054545
WAT054590
WAT054635
WAT054675
WAT054720
Accell™ Plus CM
WAT020850
WAT054500
WAT054550
WAT054595
WAT054640
WAT054680
WAT054725
Accell™ Plus QMA
WAT020820
WAT043490
WAT054535
WAT054580
WAT054620
WAT054670
WAT054710
Alumina A
WAT020825
WAT043495
WAT054540
WAT054585
WAT054625
WAT054665
WAT054715
Alumina B
WAT020830
WAT043485
WAT054530
WAT054575
WAT054630
WAT043375
WAT054705
Alumina N
WAT020840
WAT054515
WAT054560
WAT054605
WAT054650
WAT054695
WAT054740
Amino Propyl (NH2)
WAT020835
WAT054510
WAT054555
WAT054600
WAT054645
WAT054685
WAT054730
Cyano Propyl (CN)
WAT020845
WAT054520
WAT054565
WAT054610
WAT054655
WAT054690
WAT054735
Diol
mg = nominal weight of sorbent in milligrams.
See Sep-Pak® cartridge configuration and nominal weight of sorbent chart on
page 32 for specific values of each sorbent.
© 2001 Waters Corporation
35
Sample Preparation
Sep-Pak® Plus Cartridges
Box of 50
C18
tC18 (trifunctional)
C8
tC2 (trifunctional)
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT020515
WAT036810
WAT036775
WAT052720
WAT020520
WAT020525
WAT020550
WAT020545
WAT020500
WAT020505
WAT020510
WAT020535
WAT020540
WAT020530
127.00
127.00
127.00
127.00
127.00
127.00
132.00
132.00
127.00
127.00
127.00
127.00
127.00
132.00
Part No.
Price
WAT051910
WAT051900
WAT051960
WAT010910
WAT010835
WAT051800
WAT051820
WAT051810
WAT010830
WAT010823
WAT010818
134.00
134.00
134.00
140.00
140.00
134.00
134.00
134.00
134.00
134.00
140.00
Part No.
Price
WAT023501
WAT036805
WAT036770
WAT052725
WAT023537
WAT023543
WAT023531
WAT023525
WAT023549
WAT023555
WAT023561
WAT023513
WAT023507
WAT023519
114.00
114.00
114.00
114.00
114.00
114.00
120.00
120.00
114.00
114.00
114.00
114.00
114.00
120.00
Sep-Pak® Plus cartridges come in short
body (C18, tC18, C8, tC2, Accell Plus
QMA and CM, NH2, CN, Diol) and
long body (Silica, Florisil, Alumina A,B,N)
configurations. They are stackable,
allowing for increased capacity or for
the use of more than one sorbent.
Sep-Pak® Plus cartridges are compatible
with some automated sample processors.
Sep-Pak® Classic Cartridges
Box of 50
C18
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Sep-Pak® Classic cartridges also come in
short body (C18, Accell Plus QMA and
CM, NH2, CN, Diol) and long body
(Silica, Florisil, Alumina A,B,N) configurations. Waters original Sep-Pak® cartridges,
radially compressed “mini-columns”, for
versatile easy column liquid-solid extraction.
Sep-Pak® Light Cartridges
Box of 50
C18
tC18 (trifunctional)
C8
tC2 (trifunctional)
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
© 2001 Waters Corporation
36
For “sample limited” applications or
where excessive dilution is a concern,
Sep-Pak® Light cartridges contain less
sorbent than the standard Sep-Pak®
cartridge and can give higher sample
recovery with less solvent consumption
for limited sample situations.
Sample Preparation
Sep-Pak® Vac SPE Cartridges
Sample Preparation
Waters Sep-Pak® Vac solid-phase extraction columns come in a wide
range of syringe-barrel configurations, from 1 cc to 35 cc, for all your
high volume batch processing and automated sample prep needs. They
are packed with a variety of sorbent weights from 50 mg to 10 grams to
handle all your sample load requirements.
Sep-Pak® Vac cartridges provide the method reproducibility you need for
dependable analytical results and they are registered with the FDA as
Class 1 medical Devices. They are manufactured in strict compliance
with cGMP guidelines in manufacturing facilities certified to meet the
ISO 9002 standard.
Sep-Pak® Vac 1 cc Cartridges (contains 50 mg of sorbent)
Box of 100
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Cyano Propyl (CN)
Part No.
Price
WAT054955
WAT054960
WAT054965
WAT054980
WAT054985
WAT054975
107.00
107.00
107.00
95.00
107.00
151.00
For information on the availability of other sorbents, please contact your
local sales representative.
Sep-Pak® Vac 1 cc Cartridges (contains 100 mg of sorbent)
Box of 100
C18
tC18 (trifunctional)
C8
tC2 (trifunctional)
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT023590
WAT036820
WAT036785
WAT052710
WAT023595
WAT023600
WAT023625
WAT023620
WAT023575
WAT023580
WAT023585
WAT023610
WAT023615
WAT023605
115.00
115.00
115.00
115.00
105.00
115.00
169.00
169.00
115.00
115.00
115.00
169.00
169.00
169.00
Sep-Pak® Vac 3 cc Cartridges (contains 200 mg of sorbent)
Box of 50
C18
tC18 (trifunctional)
C8
Silica
Cyano Propyl (CN)
Part No.
Price
WAT054945
WAT054925
WAT054940
WAT054930
WAT054935
83.00
82.00
83.00
78.00
102.00
Sep-Pak® Vac 3 cc Cartridges (contains 500 mg of sorbent)
Box of 50
C18
tC18 (trifunctional)
C8
tC2 (trifunctional)
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT020805
WAT036815
WAT036780
WAT052715
WAT020810
WAT020815
WAT020855
WAT020850
WAT020820
WAT020825
WAT020830
WAT020840
WAT020835
WAT020845
91.00
91.00
91.00
91.00
87.00
91.00
110.00
110.00
91.00
91.00
91.00
110.00
110.00
110.00
Sep-Pak® Vac 6 cc Cartridges (contains 500 mg of sorbent)
Box of 30
C18
tC18 (trifunctional)
C8
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT043395
WAT036790
WAT054525
WAT043405
WAT054545
WAT054550
WAT054535
WAT054540
WAT054530
WAT054560
WAT054555
WAT054565
76.00
76.00
76.00
69.00
92.00
92.00
69.00
69.00
69.00
92.00
92.00
92.00
© 2001 Waters Corporation
37
Sample Preparation
Sep-Pak® Vac 6 cc Cartridges (contains 1 g of sorbent)
Box of 30
C18
tC18 (trifunctional)
C8
tC2 (trifunctional)
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Sep-Pak® Vac 35 cc Cartridges (contains 10 g of sorbent)
Part No.
Price
WAT036905
WAT036795
WAT054570
WAT052705
WAT036910
WAT043390
WAT054590
WAT054595
WAT054580
WAT054585
WAT054575
WAT054605
WAT054600
WAT054610
92.00
92.00
92.00
92.00
78.00
78.00
139.00
139.00
78.00
78.00
78.00
139.00
139.00
139.00
Sep-Pak® Vac 12 cc Cartridges (contains 2 g of sorbent)
Box of 20
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Box of 10
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT043345
WAT043350
WAT054700
WAT043355
WAT043360
WAT054720
WAT054725
WAT054710
WAT054715
WAT054705
WAT054740
WAT054730
WAT054735
131.00
133.00
133.00
78.00
78.00
185.00
185.00
78.00
78.00
78.00
185.00
185.00
185.00
WAT036915
WAT043380
WAT054615
WAT036920
WAT043385
WAT054635
WAT054640
WAT054620
WAT054625
WAT054630
WAT054650
WAT054645
WAT054655
100.00
100.00
100.00
69.00
69.00
139.00
139.00
69.00
69.00
69.00
139.00
139.00
139.00
Description
Qty
Part No.
Price
30 cc Reservoir
60 cc Reservoir
Reservoir Adapters for 1, 3
and 6 cc VAC
Reservoir Adapters for 12, 20
and 35 cc VAC
Male-Male Adapter
Male Luer Plugs
Female Luer Plugs
48
12
WAT011390
WAT024659
49.00
12.00
12
WAT054260
35.00
10
100
WAT048160
WAT024310
WAT044395
WAT044385
165.00
55.00
32.00
32.00
Part No.
Price
WAT036925
WAT043365
WAT054660
WAT036930
WAT043370
WAT054675
WAT054680
WAT054670
WAT054665
WAT043375
WAT054695
WAT054685
WAT054690
163.00
163.00
163.00
100.00
100.00
210.00
210.00
100.00
100.00
100.00
210.00
210.00
210.00
© 2001 Waters Corporation
38
Price
Sep-Pak® Cartridge Accessories
Sep-Pak® Vac 20 cc Cartridges (contains 5 g of sorbent)
Box of 20
Part No.
Sample Preparation
Sep-Pak® Vac RC robotic compatible extraction cartridges contain
the same quality and amount of sorbent as our 1 cc - 100 mg and
3 cc - 500 mg Sep-Pak® Vac cartridges which makes methods
transfer easier.
Sep-Pak® Vac RC Cartridges (contains 100 mg of sorbent)
Box of 50
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
Part No.
Price
WAT036935
WAT043410
WAT043415
WAT036940
WAT043420
WAT043465
WAT043460
WAT043450
WAT043455
WAT043445
WAT043475
WAT043470
WAT043480
78.00
78.00
78.00
69.00
69.00
105.00
105.00
69.00
69.00
69.00
105.00
105.00
105.00
Sep-Pak® Vac RC Cartridges (contains 500 mg of sorbent)
Box of 50
C18
tC18 (trifunctional)
C8
Silica
Florisil®
Accell™ Plus CM
Accell™ Plus QMA
Alumina A
Alumina B
Alumina N
Amino Propyl (NH2)
Cyano Propyl (CN)
Diol
TM
Part No.
Price
WAT036945
WAT043425
WAT043430
WAT036950
WAT043435
WAT054505
WAT054500
WAT043490
WAT043495
WAT043485
WAT054515
WAT054510
WAT054520
110.00
110.00
110.00
92.00
92.00
146.00
146.00
92.00
92.00
92.00
146.00
146.00
146.00
Visit Waters on-line
product resource today
© 2001 Waters Corporation
39
Sample Preparation
Sep-Pak® Vac RC Cartridges are labeled
with sorbent type and weight and colorcoded for easy identification. Each cartridge contains either 500 mg or 100 mg
of high quality sorbent and can be utilized by many robotic sample processors.
Use Sep-Pak® Vac RC cartridges manually or with automated or robotic
equipment. The “robotic compatible” extraction cartridges feature a
20 mL overall capacity which allows for larger conditioning /elution
volumes. They eliminate the need for adapting reservoirs to standard
size Vac cartridges making sample preparation more convenient.
Sample Preparation
The Waters Extraction Manifold for Use With
Solid-Phase Extraction Cartridges.
We are pleased to introduce a new vacuum manifold from Waters. The
new extraction manifold incorporates improvements designed to make
your solid phase extraction process easier, faster and more accurate.
The vacuum manifold has the capacity to process up to twenty samples
simultaneously. The extraction manifold has enhanced features designed
for use with conventional silica-based solid phase extraction cartridges as
well as modifications that allow you take full advantage of the unique performance characteristics of our Oasis® HLB extraction cartridges.
B
C
A
Just look at what this new manifold has to offer:
A
Precision machined Delrin® cover with alignment posts
for quick and easy alignment with test tube rack.
B
Vacuum gauge placement on cover, not in fluid path
allows for quick and easy waste removal at bottom by
vacuum.
C
Enhanced vacuum control valve designed for use with
Waters Oasis® HLB extraction cartridges, allows for a
quick and momentary rise in vacuum above the frit bubble point at the touch of a finger.
D
High purity polypropylene needle valves and needle
tips with minimum dead volume.
D: Required when using
silica-based SPE cartridges.
Extraction Manifolds
Description
20 position extraction manifold without rack (includes 20 needle tips, 25 plugs and ejector tool)
20 position extraction manifold with 13 mm x 75 mm test tube rack (includes 20 needle tips,
25 plugs and ejector tool)
20 position extraction manifold with 13 mm x 100 mm test tube rack (includes 20 needle tips,
25 plugs and ejector tool)
20 position extraction manifold with 16 mm x 75 mm test tube rack (includes 20 needle tips,
25 plugs and ejector tool)
20 position extraction manifold with 16 mm x 100 mm test tube rack (includes 20 needle tips,
25 plugs and ejector tool)
© 2001 Waters Corporation
40
Part number
Price
WAT200677
540.00
WAT200606
635.00
WAT200607
635.00
WAT200608
635.00
WAT200609
635.00
Sample Preparation
Accessories and Spare Parts for the Waters Extraction Manifolds
Description
pkg of 48
pkg of 12
pkg of 100
pkg of 12
pkg of 10
Price
WAT200685
WAT200691
WAT200686
WAT200687
WAT200688
WAT200689
WAT200690
WAT058839
WAT058851
WAT058840
WAT200678
WAT200679
WAT200680
WAT200681
WAT011390
WAT024659
WAT024310
WAT054260
WAT048160
WAT058871
WAT085114
WAT085115
45.00
10.00
210.00
140.00
190.00
99.00
5.00
8.00
8.00
6.00
95.00
95.00
95.00
95.00
49.00
12.00
55.00
35.00
165.00
120.00
705.00
595.00
Sample Preparation
Needle valves, pack of 20 (required when using silica-based SPE cartridges)
(not required for use with Oasis® HLB Extraction Cartridges)
Needle tips, pack of 20
Cover, 20 position without gauge assembly
Gauge assembly, Vacuum
Reservoir, glass with outlet valve
Outlet valve kit
Gasket for cover
Ejector tool
Luer Plugs, pack of 25
Rubber ball ring (for vacuum gauge assembly)
13 mm x 75 mm test tube rack
13 mm x 100 mm test tube rack
16 mm x 75 mm test tube rack
16 mm x 100 mm test tube rack
Reservoir, 30 cc (for Sep-Pak® Plus, Light, Vac & Classic cartridges)
Reservoir, 60 cc (for Sep-Pak® Plus, Light & Vac cartridges)
Adapter, male-male Luer (for Sep-Pak® Classic cartridges)
Adapter (to attach reservoir to 1, 3 & 6 cc Sep-Pak® Vac cartridges)
Adapter (to attach reservoir to 12, 20 & 35 cc Sep-Pak® Vac cartridges)
Reversible Vial Rack for 1 mL or 4 mL autosampler vials
Vacuum pump (110V, 60 Hz)
Vacuum pump (220V, 50 Hz)
Part No.
Accessories and Spare Parts for the Waters Vacuum Manifolds
(Older Design with Blue Lid) 12, 16 & 24 Position
Description
C-Clips
Cover, Blue Nylon, with Gasket and 12 Stopcocks
Cover, Blue Nylon, with Gasket and 16 Stopcocks
Cover, Blue Nylon, with Gasket and 24 Stopcocks
Eluent Delivery Needles, Polypropylene
Eluent Delivery Needles, Stainless Steel
Gaskets
Gaskets
Gauge and Bleed Valve Assembly
Luer Fitting, Female (Top of Cover, for Stopcocks)
Luer Fitting, Male (Bottom of Cover, for Needles)
Plastic Legs, Black
Stopcocks
Vacuum Chamber, Glass
Vacuum Chamber, Glass
Collection Rack Assembly
Holder Plate, 10 mm Test Tubes or Autosampler Vials
Holder Plate, 12 mm Test Tubes
Holder Plate, 16 mm Test Tubes
Holder Plate, Volumetric Flasks
Collection Rack Assembly
Holder Plate, 12 mm Test Tubes
Holder Plate, 16 mm Test Tubes
Collection Rack Assembly
Holder Plate, 12 mm Test Tubes
Holder Plate, 16 mm Test Tubes
Reservoirs (for Sep-Pak Plus, Light, Classic Cartridges)
Adapters (male-male) for Classic cartridges
Position Size
Quantity
Part No.
Price
12/16/24
12
16
24
12/16/24
12/16/24
12
16/24
12/16/24
12/16/24
12/16/24
12/16/24
12/16/24
12
16/24
12
12
12
12
12
16
16
16
24
24
24
12/16/24
12/16/24
12
1
1
1
12
12
2
2
1
2
2
4
12
1
1
1
1
1
1
1
1
1
1
1
1
1
48
100
WAT054822
WAT054800
WAT054801
WAT054802
WAT054808
WAT054807
WAT054803
WAT054804
WAT054799
WAT054809
WAT054810
WAT054805
WAT054806
WAT054797
WAT054798
WAT054811
WAT054812
WAT054813
WAT054814
WAT054815
WAT054816
WAT054817
WAT054818
WAT054819
WAT054820
WAT054821
WAT011390
WAT024310
20.00
150.00
235.00
265.00
12.00
39.00
8.00
14.00
95.00
4.00
4.00
10.00
15.00
85.00
115.00
85.00
35.00
35.00
35.00
35.00
105.00
50.00
50.00
105.00
50.00
50.00
49.00
55.00
© 2001 Waters Corporation
41
Sample Preparation
Waters Alliance® Filtration System
The introduction of the Alliance® Filtration System dramatically changes
the way sample filtration is performed for Alliance® HPLC systems. In only
seconds up to 24 samples can be simultaneously fitted into vials held in a
Waters 2690 Separation module’s sample carousel. The Alliance®
Filtration System eliminates the time-consuming alternative one-at-a-time
syringe filter process.
The Alliance® Filtration System is comprised of a manifold which houses
the Alliance® sample carousel and a Pall Gelman Laboratory AcroPrep™
disposable filter plate containing 24 membrane filters.
Here’s how it works:
Simply place your Alliance® carousel with empty sample vials into the
manifold. The manifold contains an alignment key which locks the
carousel into place ensuring that sample vials in the carousel line up
directly under the appropriate filter well on the AcroPrep™ filter plate. The
AcroPrep™ filter plate is placed onto the manifold. Alignment posts on the
manifold ensure that the filter wells line up directly over the appropriate
sample vial in the carousel. Pipette your samples into the filter wells on
the AcroPrep™ filter plate, apply a vacuum, and in seconds, the samples
are drawn through the AcroPrep™ filter plate into the appropriate sample
vials housed in the carousel. Once the samples are filtered, simply shut
off the vacuum source, remove and dispose of the AcroPrep™ filter plate,
cap your sample vials and you’re ready to run. For applications that
require a filter prewash step, a waste tray is included with the Alliance®
Filtration System.
AcroPrep™ filter plates for the Alliance® Filtration System are available with
0.45 µm PTFE, PVDF, Nylon, and GH Polypro (GHP) membrane filters
from Pall Gelman Laboratory.
Some advantages the Alliance® Filtration System has
over the conventional syringe filter technique
Ease-of-Use: No need to use cumbersome, individually wrapped
disposable syringes. With the Alliance® Filtration System you can use
disposable pipette tips which are less costly, easier to use and require
less bench space.
Saves Time: The conventional syringe filter technique is a one-at-a-time
filtration process. With the Alliance® Filtration System, you can simultaneously filter up to 24 samples, in only seconds, saving both time and labor.
Saves Space: Using the conventional syringe filter technique requires
significant bench space. The Alliance® Filtration System is compact and
requires much less bench space.
Affordable: When factoring in material and labor costs, the
Alliance® Filtration System is comparable in cost to the conventional
syringe filter technique.
Improves Productivity: The conventional syringe filter technique requires
25 minutes of labor per carousel. The Alliance® Filtration System improves
productivity in your lab by saving 20 minutes in labor time per carousel.
The materials of construction used in the AcroPrep™ filter plates are the
same as those used in the Pall Gelman Laboratory Acrodisc® syringe filters. If you are currently using Acrodisc® syringe filters for your sample filtration, this should eliminate the need to validate the AcroPrep™ filter plates.
Material and labor requirements
Syringe Filter Technique
Alliance® Filtration System
24 disposable syringes
24 disposable pipette tips
24 syringe filters
1 AcroPrep™ filter plate
25 minutes
5 minutes
© 2001 Waters Corporation
42
Sample Preparation
Compatibility Chart for AcroPrep™ Filter Plates
Alliance Filtration Manifold (complete)
®
Solvent
GHP
Nylon
PVDF
PTFE
Acetone
Acetonitrile
Acetic acid, glacial
n-Butanol
Chloroform
Dioxane
Dimethyl formamide
Dimethyl sulfoxide
Ethanol
Ethyl acetate
Ethyl ether
Freon TF
Solvent
Hydrochloric acid (1N)
Hexane, dry
Methanol
Methylene chloride
Methyl ethyl ketone
N-Methylpyrrolidone
Isopropanol
Sodium hydroxide (3N)
Tetrahydrofuran
Tetrahydrofuranl/water (50/50)
Toluene
Water
R
R
R
R
R
R
R
R
R
R
R
R
GHP
R
R
R
R
R
R
R
R
R
R
R
R
R
R
NR
R
R
*
R
R
R
R
R
R
Nylon
NR
R
R
R
NR
R
R
LR
R
R
R
R
NR
R
R
R
R
R
NR
NR
R
R
R
R
PVDF
R
R
R
R
NR
NR
R
NR
NR
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
PTFE
R
R
R
R
R
R
R
R
R
R
R
R
Quantity
Part No.
Price
1
289000159
750.00
1
3/pkg
3/pkg
3/pkg
1
1
1
5 carousels
700000231
700000232
700000233
700000234
700000235
WAT085114
WAT085115
WAT270328
75.00
15.00
6.00
22.00
25.00
705.00
595.00
205.00
186000154
186000155
186000156
186000157
460.00
460.00
460.00
460.00
Sample Preparation
Description
Spare Parts and Accessories
R = Resistant LR = Limited Resistance NR = Not Resistant * = Insufficient Data
Glass Housing for Filtration Manifold
O-Ring Kit
Rubber Feet
Manifold Alignment Post
Waste Tray
Vacuum Pump 110 V, 60Hz
Vacuum Pump 220V, 50Hz
Alliance® Carousel Kit
AcroPrep™ Filter Plates for the Alliance® Filtration System
AcroPrep™ GHP Filter Plates, 0.45 µm
AcroPrep™ Nylon Filter Plates, 0.45 µm
AcroPrep™ PTFE Filter Plates, 0.45 µm
AcroPrep™ PVDF Filter Plates, 0.45 µm
10/pkg
10/pkg
10/pkg
10/pkg
2 mL (12 x 32 mm) Crimp Top and Snap Cap Vials and Accessories
(6.0 mm neck opening) Recommended for use with the Alliance® Filtration System
12 x 32 mm clear glass vial, crimp top
12 x 32 mm amber glass vial, crimp top
Polyurethane cap (0.44 mm with star)
12 x 32 mm clear glass vial, snap cap design,
glass is Type 1, Class A
12 x 32 mm amber glass vial, snap cap design,
glass is Type 1, Class B
One-step plastic cap and septa (PTFE)
Alliance® total recovery vial
100/pkg
100/pkg
100/pkg
100/pkg
WAT094222
WAT094223
WAT094224
WAT094219
16.00
18.70
26.00
15.00
100/pkg
WAT094220
17.50
100/pkg
100/pkg
186000328
186000234
18.50
98.00
Test Methods: The data presented in this chart is a compilation of testing by Pall
Gelman Laboratory with certain chemicals, manufacturer’s data, or compatibility
recommendations from the Compass Corrosion Guide, by Kenneth M. Pruett. This
data is intended to provide expected results when filtration devices are exposed to
chemicals under static conditions for 48 hours at 25 °C, unless otherwise noted.
This chart is intended only as a guide. Accuracy cannot be guaranteed.
Users should verify chemical compatibility with a specific filter under under actual
use conditions. Chemical compatibility with a specific filter, under actual use conditions, is affected by many variables, including temperature, pressure, concentration,
purity, and various chemical combinations which present complete accuracy.
© 2001 Waters Corporation
43
Sample Preparation
Pall Gelman Laboratory Sample and
Solvent Filtration Products from Waters
Membrane Choices
Membrane Type (PTFE, GHP, PVDF, Nylon) — Typically, the first step
in selecting a suitable filter is to choose a membrane that is chemically
compatible with the sample (see chart opposite). For aqueous samples,
it is preferable to use a hydrophilic membrane (GHP, PVDF or Nylon)
versus a hydrophobic membrane (PTFE). With proteinaceous samples,
a low protein binding filter (GHP or PVDF) is preferable to other
hydrophilic filters (Nylon).
GHP Acrodiscs
Filtration of HPLC samples and solvents is a preventative maintenance
procedure that saves your chromatography lab time and money. Filtration
provides immediate protection for the components of your HPLC
instrumentation by prolonging column life, minimizing downtime, and
improving both the precision and accuracy of your data. Filtering samples
and mobile phase prior to use decreases downtime by eliminating
sample/solvent particulates that may contribute to:
• erratic pressure fluctuations
• noisy base lines
• damage to pump pistons and seals, injector
valves and plugged injector needles
• noise in the detector
• decreased column lifetime
• HPLC certified
• Hydrophilic polypropylene membrane offers excellent
chemical compatibility and minimizes extractables
• Suitable for aqueous and organic solvents
• Low protein binding
Nylon Acrodiscs
• HPLC certified
• Save time, no pre-wetting
• Hydrophilic nylon membrane offers broad
chemical compatibility
Acrodisc® LC (PVDF)
• HPLC certified
• Hydrophilic PVDF membrane
• Filters both aqueous and organic solvent solutions
Pall Gelman Laboratory filters have been Certified for Compliance.
That means they’ve been designed and developed to assist customers
in complying with their regulatory and quality objectives. We have more
membrane types such as hydrophilic polypropylene for broad chemical
compatibility and low protein binding. Acrodiscs come in a variety of
sizes, including the new minispike which is designed for filtering small
volumes with minimum hold-up volume into narrow openings such as
autosampler vials. The Acrodisc® membrane is housed in high purity
polypropylene, resulting in exceptionally low extractable levels.
Acrodisc® CR (PTFE)
• HPLC certified
• Excellent chemical compatibility
• Used for aggressive and organic solvents
Ion Chromatography (IC) Acrodiscs
• IC certified to ensure low ionic backgrounds
• Recommended for aqueous samples only
GHP Acrodisc® GF and Nylon Acrodisc® GF
•
•
•
•
Filter Design Choices
Pall Gelman Laboratory’s broad range of device sizes assures complete
sample filtration with minimal sample loss. In addition, many Acrodisc®
syringe filters offer the option of a minispike outlet, rather than the
traditional male luer outlet. The minispike outlet, available on 13 mm
PTFE, Nylon, GH Polypro and PVDF Acrodisc® syringe filters, allows
for easy dispensing into autosampler vials.
Designed with a glass fiber prefilter over membrane
Maximum throughputs for hard-to-filter samples
HPLC certified
Binder-free borosilicate glass fiber prefilter
Glass Fiber Acrodiscs
• 100% binder-free borosilicate glass fiber filter
• Can be used alone or in series with another Acrodisc®
• deal for prefiltration of 5-100 milliliters of serum
Supor® (PES)
• Hydrophilic polyethersulfone
• For biological, pharmaceutical or sterilizing sample
• Can be gamma sterilized or autoclaved
4mm
PP
Male Luer
13mm
PP
Male Luer
13mm
PP
Minispike
25mm
PP
Male Luer
PP = Polypropylene housing
Pore Size (0.2 and 0.45 µm) — The filter pore size is usually a
personal preference of the chromatographer, but relates to the column
packing material. Since there are many different packing materials, a
general guideline is that most customers find that the 0.45 µm filter meets
their requirements, although chromatographers are increasingly moving
toward using 0.2 µm filters.
© 2001 Waters Corporation
44
Sample Preparation
Water
Toluene
Tetrahydrofuran/water (50/50)
Tetrahydrofuran
Sodium hydroxide (5N)
Isopropanol
N-Methylpyrrolidone
Methyl ethyl ketone
Methylene chloride
Methanol
Hexane, dry
Hydrochloric acid (1N)
Freon TF
Ethyl ether
Ethyl acetate
Ethanol
Dimethyl sulfoxide
Dimethyl formamide
Dioxane
Chloroform
n-Butanol
Acetic acid, glacial
Acetoniltrile
= RESISTANT
= LIMITED RESISTANCE
= NOT RESISTANT
= INSUFFICIENT DATA
GH Polypro Syringe Filters
GHP Acrodisc® 13 (13 mm)
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
GHP Acrodisc® (25 mm)
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
GHP Acrodisc® GF (25 mm)
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
PTFE Syringe Filters
Acrodisc® 4CR PTFE (4 mm)
R*
R
R
R
LR
R
R*
R*
R
R*
R
R
R
R
R
LR
R*
R*
R
LR
LR
•
LR*
R
Acrodisc® 13CR PTFE (13 mm)
R*
R
R
R
R
R
R*
R*
R
R*
R
R
R
R
R
R
R*
R*
R
R
R
R
R*
R
Acrodisc® CR PTFE (25 mm)
R*
R
R
R
R
R
R*
R*
R
R*
R
R
R
R
R
R
R*
R*
R
R
R
R
R*
R
Acrodisc® LC13 PVDF (13 mm)
NR*
R
R
R
R
R
NR* NR*
R
R*
R
R
R
R
R
R
NR* NR*
R
NR
R
R
R*
R
Acrodisc LC PVDF (25 mm)
NR*
R
R
R
R
R
NR* NR*
R
R*
R
R
R
R
R
R
NR* NR*
R
NR
R
R
R*
R
Nylon Acrodisc® 4 (4 mm)
R*
R
R
R
NR
•
R*
R*
R
R*
NR
R
NR
R
R
NR
R*
R*
R
R
NR
LR
R*
R
Nylon Acrodisc® 13 (13 mm)
R*
R
R
R
NR
•
R*
R*
R
R*
NR
R
NR
R
R
NR
R*
R*
R
R
NR
LR
R*
R
Nylon Acrodisc® (25 mm)
R*
R
R
R
NR
•
R*
R*
R
R*
NR
R
NR
R
R
NR
R*
R*
R
R
NR
LR
R*
R
Nylon Acrodisc GF (25 mm)
R*
R
R
R
NR
•
R*
R*
R
R*
NR
R
NR
R
R
NR
R*
R*
R
R
NR
LR
R*
R
PVDF Syringe Filters
®
Nylon Syringe Filters
®
Ion Chromatography Syringe Filters
IC Acrodisc® (13 mm & 25 mm)
NR
LR
NR
R
NR
•
NR
NR
•
LR
R
LR
•
LR
R
NR
•
NR
•
•
NR
•
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
LR
R
R
R
R
NR
NR
NR
R
NR
NR
R
LR
NR
R
NR
NR
NR
R
R
NR
NR
NR
R
R
Glass Fibre Syringe Filters
GF Acrodisc®
R
Acrylic Copolymer Syringe Filters
Non-sterile Acrodisc® (25 mm)
NR
NR
NR
R
NR
Disc Filters
GH Polypro
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
NR
R
R
R
R
LR
NR
NR
R
R
R
R
R
R
R
R
LR
NR
R
NR
LR
•
R
R
Nylaflo (Nylon)
R
R
NR
R
NR
R
R
R
R
R
R
LR
NR
•
LR
NR
NR
R
R
R
R
R
NR
R
TF (PTFE)
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
FP Verciel (PVDF)
Note:
R = Resistant
LR = Limited Resistance
NR = Not Resistant
No significant change was observed in flow rate or bubble point of the membrane.
Moderate changes in physical properties or dimension of
the membrane were observed. The filter may be suitable
for short term, non-critical use at room temperature.
The membrane is basically unstable. In most cases, extensive shrinkage or swelling occurs. The filter may gradually
weaken or partially dissolve after extended exposure.
* UV absorbance was set at 254 nm
© 2001 Waters Corporation
45
Sample Preparation
R
LR
NR
•
Acetone
Filter Design and Membrane Choices
Sample Preparation
Part Number Cross Reference
Pall Gelman Laboratory
Description
Whatman
Part No.
Millipore®
Part No.
Waters
Part No.
PTFE, 4 mm, 0.45 µm
PTFE, 13 mm, 0.2 µm
PTFE, 13 mm, 0.45 µm
PTFE, 25 mm, 0.2 µm
PTFE, 25 mm, 0.45 µm
PVDF, 13 mm, 0.45 µm
PVDF, 25 mm, 0.45 µm
GHP, 13 mm, 0.45 µm
GHP, 25 mm, 0.45 µm
Nylon, 4 mm, 0.45 µm
Nylon, 13 mm, 0.2 µm
Nylon, 13 mm, 0.45µm
Nylon, 25 mm, 0.2 µm
Nylon, 25 mm, 0.45 µm
Supor®, 25 mm, 0.45 µm
Supor®, 25 mm, 0.2 µm
PVDF, 47 mm, 0.45 µm
PVDF, 47 mm, 0.2 µm
Nylon, 47 mm, 0.45 µm
Nylon, 47 mm, 0.2 µm
PTFE, 47 mm, 0.45 µm
PTFE, 47 mm, 0.2 µm
GHP, 47 mm, 0.45 µm
Supor®, 47 mm, 0.2 µm
Supor®, 47 mm, 0.45 µm
Supor®, 13 mm, 0.45 µm
Glass Filter Holder
Forceps
300 mL Funnel
Glass Base, Tabulated Cap
Ground Joint Flask
Thick Glass Prefilter
AVE Glass Prefilter
6783-0404
6783-1302
6783-1304
6784-2502
6784-2504
6792-1304
—
6784-1304
6785-2504
6790-0404
6790-1302
6790-1304
—
—
6780-2504
6780-2502
—
—
7404-004
7402-004
7585-004
7582-004
7002-0447
—
—
—
—
—
—
—
—
—
—
SJFHL04NS
SJFG013NS
SJFH013NS
SLFG025NS
SLSR025NS
SJHV013NS
SLHV025NS
SJFH013NS
SLCR025NS
SJHV004NS
—
—
—
—
SLHA025OS
SLGS025OS
HVLP04700
—
—
—
FHLP04700
FGLP04700
FHLC04700
GSTF04700
HATF04700
HATF01300
XX1504700
XX6200006
XX1004704
XX1504702
XX1504705
AP2501000
AP4001000
WAT200508
WAT200506
WAT200502
WAT200504
WAT200500
WAT200512
WAT200510
WAT200516
WAT200514
WAT200526
WAT200524
WAT200520
WAT200522
WAT200518
WAT200528
WAT200529
WAT200530
WAT200531
WAT200532
WAT200533
WAT200534
WAT200535
WAT200537
WAT200539
WAT200538
WAT200540
WAT200543
WAT200544
WAT200545
WAT200546
WAT200547
WAT200541
WAT200542
© 2001 Waters Corporation
46
Sample Preparation
Syringe Filters with PTFE Membrane
Quantity
Diameter
Pore Size
Acrodisc® CR
pkg/50
case/200
pkg/100
case/300
pkg/250
case/750
pkg/100
case/300
pkg/50
case/200
pkg/100
case/300
pkg/100
case/300
25 mm
25 mm
13 mm
13 mm
4 mm
4 mm
13 mm
13 mm
25 mm
25 mm
13 mm
13 mm
13 mm
13 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
Acrodisc® CR Minispike
Acrodisc® CR
Acrodisc® CR Minispike
Part No.
Price
WAT200500
WAT200501
WAT200502
WAT200503
WAT200508
WAT200509
WAT200558
WAT200559
WAT200504
WAT200505
WAT200506
WAT200507
WAT200556
WAT200557
120.00
398.00
158.00
408.52
271.00
765.06
166.00
419.64
120.00
398.00
158.00
408.52
166.00
419.64
Sample Preparation
Description
Syringe Filters with PVDF Membrane
Description
Quantity
Diameter
Pore Size
Acrodisc® LC
pkg/50
case/200
pkg/100
case/300
pkg/100
case/300
pkg/100
case/300
pkg/50
case/200
pkg/100
case/300
25 mm
25 mm
13 mm
13 mm
13 mm
13 mm
13 mm
13 mm
25 mm
25 mm
13 mm
13 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
Acrodisc® LC Minispike
Acrodisc® LC
Acrodisc® LC Minispike
Part No.
Price
WAT200510
WAT200511
WAT200512
WAT200513
WAT200560
WAT200561
WAT200806
WAT200807
WAT200808
WAT200809
WAT200804
WAT200805
110.00
382.43
150.00
398.00
155.00
407.52
155.00
409.00
110.00
385.51
158.00
409.00
Syringe Filters with GHP Membrane
Description
Quantity
Diameter
Pore Size
GHP Acrodisc®
pkg/50
case/200
pkg/50
case/200
pkg/100
case/300
pkg/50
case/200
pkg/100
pkg/300
25 mm
25 mm
25 mm
25 mm
13 mm
13 mm
25 mm
25 mm
13 mm
13 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
GHP Acrodisc® GF
GHP Acrodisc® Minispike
GHP Acrodisc®
GHP Acrodisc® Minispike
Part No.
Price
WAT200514
WAT200515
WAT200802
WAT200803
WAT200516
WAT200517
WAT097964
WAT097965
WAT097962
WAT097963
120.00
382.00
117.00
405.33
165.00
420.00
120.00
398.00
165.00
420.00
© 2001 Waters Corporation
47
Sample Preparation
Syringe Filters with Nylon Membrane
Description
Nylon Acrodisc
®
Nylon Acrodisc® GF
Nylon Acrodisc® Minispike
Nylon Acrodisc®
Nylon Acrodisc® Minispike
Quantity
Diameter
Pore Size
pkg/50
case/200
pkg/100
case/300
pkg/250
case/750
pkg/50
case/200
pkg/100
case/300
pkg/50
case/200
pkg/100
case/300
pkg/100
case/300
25 mm
25 mm
13 mm
13 mm
4 mm
4 mm
25 mm
25 mm
13 mm
13 mm
25 mm
25 mm
13 mm
13 mm
13 mm
13 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
Part No.
Price
WAT200518
WAT200519
WAT200520
WAT200521
WAT200526
WAT200527
WAT200800
WAT200801
WAT200564
WAT200565
WAT200522
WAT200523
WAT200524
WAT200525
WAT200562
WAT200563
118.00
384.00
149.00
408.96
265.00
736.81
116.00
401.86
155.00
407.52
118.00
384.00
149.00
408.96
155.00
407.52
Part No.
Price
WAT200528
WAT200529
86.00
86.00
Part No.
Price
WAT200812
WAT200813
WAT200816
WAT200817
WAT200810
WAT200811
WAT200814
WAT200815
169.00
435.00
122.00
419.00
169.00
435.00
122.00
419.00
Part No.
Price
WAT200818
WAT200819
63.00
226.00
Sterile Syringe Filters with Polyethersulfone Membrane
Description
Quantity
Supor Acrodisc
®
pkg/50
®
Diameter
Pore Size
25 mm
0.45 µm
0.2 µm
Syringe Filters with Supor® Membrane
Description
Quantity
Diameter
Pore Size
IC Acrodisc
pkg/100
case/300
pkg/50
case/200
pkg/100
case/300
pkg/50
case/200
13 mm
13 mm
25 mm
25 mm
13 mm
13 mm
25 mm
25 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
®
Syringe Filters with Glass Fiber Membrane
Description
Glass Fiber Acrodisc
®
Quantity
Diameter
Pore Size
pkg/50
case/200
25 mm
25 mm
1.0 µm
1.0 µm
© 2001 Waters Corporation
48
Sample Preparation
Special Bulk Packaging (1000 devices)
Description
Diameter
Qty
Part No.
Price
0.45
0.45
0.2
0.2
0.2
0.45
1000
1000
1000
1000
1000
1000
WAT200820
WAT200821
WAT200822
WAT200823
WAT200824
WAT200825
998.00
772.00
998.00
772.00
772.00
772.00
25 mm
13 mm
13 mm
13 mm
25 mm
0.45
0.45
0.45
0.2
0.2
1000
1000
1000
1000
1000
WAT200826
WAT200827
WAT200828
WAT200838
WAT200839
998.00
772.00
772.00
772.00
998.00
25 mm
25 mm
13 mm
0.45
0.45
0.45
1000
1000
1000
WAT058853
WAT200829
WAT200830
1,050.00
1,008.00
780.00
Sample Preparation
Pore Size
Syringe Filters with PTFE Membrane
Acrodisc® CR
Acrodisc® CR
Acrodisc® CR
Acrodisc® CR
Acrodisc® CR Minispike
Acrodisc® CR Minispike
25 mm
13 mm
25 mm
13 mm
13 mm
13 mm
Syringe Filters with PVDF Membrane
Acrodisc® LC
Acrodisc® LC
Acrodisc® LC Minispike
Acrodisc® LC Minispike
Acrodisc® LC
Syringe Filters with GHP Membrane
GHP Acrodisc® GF
GHP Acrodisc®
GHP Acrodisc® Minispike
Syringe Filters with Nylon Membrane
Nylon Acrodisc®
Nylon Acrodisc®
Nylon Acrodisc®
Nylon Acrodisc®
Nylon Acrodisc® Minispike
Nylon Acrodisc® Minispike
Nylon Acrodisc® GF
25 mm
13 mm
25 mm
13 mm
13 mm
13 mm
25 mm
0.45
0.45
0.2
0.2
0.2
0.45
0.45
1000
1000
1000
1000
1000
1000
1000
WAT200831
WAT200832
WAT200833
WAT200834
WAT200835
WAT200836
WAT200846
998.00
772.00
998.00
772.00
772.00
772.00
1,040.00
Glass Fibre Acrodisc®
Versapor Acrodisc®
IC Acrodisc®
IC Acrodisc®
IC Acrodisc®
IC Acrodisc®
25 mm
25 mm
13 mm
25 mm
13 mm
25 mm
1
0.8
0.45
0.45
0.2
0.2
1000
1000
1000
1000
1000
1000
WAT200840
WAT200841
WAT200842
WAT200843
WAT200844
WAT200845
954.00
1,050.00
806.00
1,060.00
806.00
1,060.00
TM
Visit Waters on-line
product resource today
© 2001 Waters Corporation
49
Sample Preparation
Solvent Filtration Membranes
Description
Quantity
Diameter
Pore Size
PVDF Filter
Nylon Filter
PTFE Filter
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
pkg/100
47 mm
47 mm
47 mm
13 mm
47 mm
47 mm
13 mm
47 mm
47 mm
47 mm
47 mm
10 mm
10 mm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.45 µm
0.2 µm
0.2 µm
0.2 µm
0.2 µm
1.0 µm
1.0 µm
GH Polypro Filter
Supor® (PES) Filter
PVDF Filter
Nylon Filter
PTFE Filter
Supor® (PES) Filter
Thick Glass Filter
A/E Glass Filter
Part No.
Price
WAT200530
WAT200532
WAT200534
WAT200536
WAT200537
WAT200538
WAT200540
WAT200531
WAT200533
WAT200535
WAT200539
WAT200541
WAT200542
86.00
86.00
231.00
140.00
92.00
82.00
51.00
86.00
86.00
231.00
82.00
20.00
22.00
Solvent Filtration Apparatus
The 300 mL capacity 47 mm Glass Filter Funnel and 1 litre capacity
47 mm Glass Funnel/Support Assembly are ideal for vacuum filtration of
liquids and degassing of HPLC solvent and mobile phases. The 100%
borosilicate glass construction assures resistance to even the most
aggressive solvents.
Description
All Glass Filter Holder 47mm, complete
Forceps, SS
Funnel, 300 mL
Glass Base, tabulated cap
Ground Joint Flask
Swinney Holder
Part No.
Price
WAT200543
WAT200544
WAT200545
WAT200546
WAT200547
WAT200566
400.00
20.00
73.00
165.00
113.00
55.00
© 2001 Waters Corporation
50