Waters Certified Sep-Pak User Manual

[ Care and Use ManUal ]

CeRtIFIeD seP-PAK CARtRIDGes

Contents

I. IntRoDUCtIon

II. UsInG CeRtIFIeD seP-PAK CARtIDGes

III. stRAteGIes FoR soLID-PHAse eXtRACtIon

IV. FACtoRs AFFeCtInG sePARAtIon eFFICIenCY

V. stoRAGe AnD DIsPosAL oF UseD CARtRIDGes

VI. oRDeRInG InFoRmAtIon

VII. ADDItIonAL InFoRmAtIon

VII. LIteRAtURe ReFeRenCes

I. IntRoDUCtIon

Waters Certified Sep-Pak® cartridges are convenient, reproducible,

disposable solid-phase extraction (SPE) devices for sample

preparation. Certified Sep-Pak cartridges 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.

Certified Sep-Pak cartridges are available in many styles to

accommodate manual and automated solid-phase extraction and are

available in 3 cc and 6 cc formats in the following chemistries:

Reversed-Phase: C

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

18

®

Certified Sep-Pak Cartridges 1

[ Care and Use ManUal ]

Load Sample

(Black)

Step

Elute 1

Step

Elute 2

One cartridge can separate all three dyes

Step

Elute 3

NOTE: Different
strength solvents
can be used to
separate the dyes.

Stationary

Phase

Particles

II. UsInG CeRtIFIeD seP-PAK CARtRIDGes

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

Each separation is different and not all steps may be required for

your application.

1. Prepare sample: T he sample can be applied to the Certified 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.

2. Condition/equilibrate cartridge: A conditioning step is required for

reversed-phase sorbents (e.g., C

). Use a strong solvent to wet the

18

stationary phase. Conditioning is followed by an equilibration step

using a weak solvent.

3. Load sample: T he sample is loaded onto the cartridge.

4. Wash: The wash step removes interferences while retaining the analyte.

If weakly retained interferences are present in the sample matrix, wash

with an appropriate weak solvent.

5. Elute: Use an appropriate stronger solvent to elute the analyte while

retaining more strongly bound interferences.

III. stRAteGIes FoR soLID-PHAse eXtRACtIon

a. Retention-Cleanup-Elution Strategy

As the sample is loaded onto the cartridge, the analytes of interest are

retained by the sorbent. If needed, an optimized series of washes are

used to remove matrix interference from the cartridge. A strong solvent

is used to elute the analytes from the cartridge. Sample enrichment

results when the final elution volume is smaller than the load volume.

b. Pass-Through Cleanup Strategy

Pass-through cleanup methods optimize matrix retention while the

analytes of interest pass-through the cartridge unretained. No sample

enrichment occurs during the solid-phase extraction (SPE) step.

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 desir-

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

that is incompatible with the analytical method.

Certified Sep-Pak Cartridges 2

1. Sample is passed through sorbent and collected

• No sample enrichment

2. Matrix interferences are retained on sorbent

Pass-Through