Waters Sep-Pak Cartridges and Plates User Manual

[ Care and Use ManUal ]

seP-PAK CArtrIdGes And PLAtes

Waters Sep-Pak® cartridges and 96-well plates are convenient,

reproducible, disposable solid-phase extraction (SPE) devices

for sample preparation. Sep-Pak cartridges and plates are

manufactured in a Waters ISO 13485 and ISO 9002-registered

facility in compliance with cGMP guidelines for the U.S. Food and

Drug Administration for Class 1 Medical Devices.

Contents

I. IntroduCtIon

II. usInG seP-PAK CArtIdGes And PLAtes

III. strAteGIes For soLId-PHAse eXtrACtIon

a. Retention-Cleanup-Elution Strategy

b. Pass-Through Cleanup Strategy

IV. stePs oF An sPe ProCedure

V. storAGe And dIsPosAL oF used CArtrIdGes

VI. AddItIonAL InForMAtIon

VII. LIterAture reFerenCes

Sep-Pak Cartridges and Plates 1

[ Care and Use ManUal ]

I. IntroduCtIon

Sep-Pak devices are available in many styles to accommodate

manual and automated solid-phase extraction. Devices are available

in convenient Plus, Light, Classic, Vac, Vac RC, and 96-well plate

formats.

Available Sorbent Chemistries:

Reversed Phase: C18, tC18, C8, tC

Normal Phase: Silica, Alumina A, Alumina B, Alumina N, Florisil

2

®

Ion Exchange and Mixed Mode: Aminopropyl, PSA, Accell™ Plus

QMA, Accell Plus CM, Cyanopropropyl, Diol

Speciality Products: Porapak® RDX, DNPH-Silica, XPoSure™, Ozone

Scrubber, Dry, PS2, AC2, Carbon Black/Aminopropyl

II. usInG seP-PAK CArtrIdGes And 96-WeLL PLAtes

Below is a five step guideline for using Sep-Pak cartridges. Each separation

is different and not all steps may be required for your application. Typical

strong and weak solvents used in the sample preparation steps are listed in

Table 1 below.

Table 1: Typical strong and weak solvents used in the

sample preparation.

Separation Mode Typical Weak

Solvents

Reversed Phase Water or bufer.

May contain low
concentrations of organics,
such as methanol.

Typical Strong

Solvents

Acetonitrile,
methanol, or
aqueous/organic
mixture

Prepare sample: The sample can be applied to the Sep-Pak Cartridge in

either liquid or a gas phase. If the sample is a solid, it must be dissolved or

extracted prior to loading.

Condition/Equilibrate Cartridge: A conditioning step is required for

reversed-phase sorbents (e.g., C18). Use a strong solvent to wet the station-

ary phase. Conditioning is followed by an equilibration step using a weak

solvent.

Load Sample: The sample is loaded onto the cartridge.

Wash: The wash step removed interferences while retaining the analyte. If

weakly retained interferences are present in the sample matrix, wash with an

appropriate weak solvent.

Elute: Use an appropriate stronger solvent to elute the analyte while retain-

ing more strongly bound interferences.

III. strAteGIes For soLId-PHAse eXtrACtIon

Purification: Use as a “chemical filter” to retain the component of interest

while interferences elute, or to retain the interferences while the analyte

passes through unretained.

Trace Enrichment or Concentration: Use to concentrate an analyte from

a dilute solution. This is done when the analyte concentration is below the

detection limit of the analytical method.

Fractionation: Use with a step gradient of increasing solvent strength to

selectivity elute and isolate analytes based on differences in polarity.

Solvent Exchange: Use to adsorb the analyte and elute with a

desirable solvent. This is helpful of the sample is dissolved in a solvent

that is incompatible with the analytical method.

Normal Phase Hexane, toluene Methylene chloride,

methanol

Weak Cation
Exchange

Strong Anion
Exchange

Sep-Pak Cartridges and Plates 2

Low ionic strength
buffer, pH >4

Low ionic strength
buffer, pH <8

High ionic strength
buffer, or pH <2

High ionic strength
buffer, or pH >10