ENVIRONMENTALENVIRONMENTALANALYSISANALYSISI. Solid Phase ExtractionI. Solid Phase Extraction

ENVIRONMENTALENVIRONMENTALANALYSISANALYSISI. Solid Phase ExtractionI. Solid Phase Extraction

step-by-step

method development

Goal of Six-Step ModeGoal of Six-Step ModeGoal of Six-Step ModeGoal of Six-Step Mode

Isolation

Purification

Trace enrichment

Six-Step Method DevelopmentSix-Step Method DevelopmentSix-Step Method DevelopmentSix-Step Method Development

Compile known parameters

Tech Support Form

Follow step-by-step guide

Fill in method development form

Technical Support FormTechnical Support FormTechnical Support FormTechnical Support Form

Application support request form

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__Address:_______________________________________________________________________________________________________________________________________________________________________________________________________________Postal/Zipcode:_________________Phone:_________________________

ext:___________________________

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In order to provide a solution to your SPE application problem wewouldappreciate you taking the time to complete as much as possible of the twosided application request form. Our past experience in SPE consultingworkhas clearly indicated that the chances of a successful conclusion increase inline with the amount of informationsupplied!If for reasons of confidentiality you are unable to disclose the structuresofthe compounds to be extracted, a representative ofthe

Consultancy Divisionof IST would be willing to sign a suitable non-disclosure agreement inorderto progress with solving your SPE problem. Please contact yourISTDistributor for furtherinformation.

1 What is/are the compounds to be extracted?

_____________________________________________________________What are the Merck Index (12th edn.) reference numbers of the

analytes(s)(if any)?

_____________________________________________________________

Compound Structure(s)

Customer ContactInformation

Please fax or mail a photocopy of thisApplication Support Request Form toyour IST Distributor using the contactinformation on the back cover ofthecatalogue.

The Analyte(s)

13

Pertinent DataPertinent DataPertinent DataPertinent Data

The analyte(s)

The matrix

The analytical method

Existing method(s)

The AnalyteThe AnalyteThe AnalyteThe Analyte

Structure pKa(s) Concentration Sample volume Solubility Stability

The MatrixThe MatrixThe MatrixThe Matrix

Type of matrix

pH and ionic strength

Interferences

The Analytical MethodThe Analytical MethodThe Analytical MethodThe Analytical Method

Final technique

Compatible solvents

HPLC conditions

Existing MethodExisting MethodExisting MethodExisting Method

Liquid-liquid method

Current SPE method details

Data from current method

Method Development ExampleMethod Development ExampleMethod Development ExampleMethod Development Example

Pollution Unlimited1224 Sunnyside DriveElution, Alaska

(907) 663-3217 573(907) 663-3218

Dr. Anna Light

The AnalyteThe AnalyteThe AnalyteThe Analyte

The Analyte (cont.)The Analyte (cont.)The Analyte (cont.)The Analyte (cont.)

The MatrixThe MatrixThe MatrixThe Matrix

The Analytical MethodThe Analytical MethodThe Analytical MethodThe Analytical Method

Other InformationOther InformationOther InformationOther Information

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

The “Form”The “Form”The “Form”The “Form”Sample name _________________

Analyte of Interest ________________

Sample size ____________________

Sorbent Bed Mass _______ Cartridge volume_____

Sorbent phases to try 1) _____ 2) _____ 3) _____ 4) _____

1. Sample pretreatment ___________________________

2. Condition column with ____ mL of _________ at ___ mL/min

3. Equilibrate column with ____ mL of _________ at ___ mL/min.

4. Load sample at ______ mL per minute.

5. Remove interference with ____ mL of _________ at ___ L/min.

Drying step _____ minutes

6. Elute analyte with ____ mL of _________ at ___ mL/min.

Remember to include soak step if possible!

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Sample Size DeterminationSample Size DeterminationSample Size DeterminationSample Size Determination

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Sorbent CapacitySorbent CapacitySorbent CapacitySorbent Capacity

Hydrophobic phases: 1- 5% of Sorbent Mass

Ion Exchangers0.3 to 0.6 milli-equivalents per gram

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Select a MechanismSelect a MechanismSelect a MechanismSelect a Mechanism

Select a MechanismSelect a MechanismSelect a MechanismSelect a Mechanism

Select a MechanismSelect a MechanismSelect a MechanismSelect a Mechanism

Select a MechanismSelect a MechanismSelect a MechanismSelect a Mechanism

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Selecting a SorbentSelecting a SorbentSelecting a SorbentSelecting a Sorbent

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

A Reminder...A Reminder...A Reminder...A Reminder...

Sample PretreatmentSample PretreatmentSample PretreatmentSample Pretreatment

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Conditioning and Conditioning and EquilibrationEquilibrationConditioning and Conditioning and EquilibrationEquilibration

Conditioning and Conditioning and EquilibrationEquilibrationConditioning and Conditioning and EquilibrationEquilibration

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Sample Loading RateSample Loading RateSample Loading RateSample Loading Rate

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Removing InterferencesRemoving InterferencesRemoving InterferencesRemoving Interferences

Removing InterferencesRemoving InterferencesRemoving InterferencesRemoving Interferences

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Drying TimeDrying TimeDrying TimeDrying Time

Filling in the FormFilling in the FormFilling in the FormFilling in the Form

Following the GuideFollowing the GuideFollowing the GuideFollowing the Guide

Selecting an Elution SolventSelecting an Elution SolventSelecting an Elution SolventSelecting an Elution Solvent

Elution Solvent VolumeElution Solvent VolumeElution Solvent VolumeElution Solvent Volume

The Completed FormThe Completed FormThe Completed FormThe Completed Form

What if Low Recovery?What if Low Recovery?What if Low Recovery?What if Low Recovery?

Losses during rinse step

Analyte breakthrough

Poor elution

Losses During Losses During Rinse StepRinse StepLosses During Losses During Rinse StepRinse Step

Check rinse for presence of

analyte

Analyte BreakthroughAnalyte BreakthroughAnalyte BreakthroughAnalyte Breakthrough

Stack Columns Elute Separately

Analyte BreakthroughAnalyte BreakthroughAnalyte BreakthroughAnalyte Breakthrough

Check pH and ionic strength at

each step of extraction procedure

Select phase offering greater

interaction

Impact of pHImpact of pHImpact of pHImpact of pH

Analyte loaded at pH=2

Analyte loaded at pH=7

Selection of Bonded PhaseSelection of Bonded PhaseSelection of Bonded PhaseSelection of Bonded Phase

Analyte well retained on C8

Analyte poorly retained on C2

Analyte well retained on C18(EC)

Poor ElutionPoor ElutionPoor ElutionPoor Elution

Select stronger elution solvent pH, ionic strength, % organics

Select less retentive sorbent

Include a soak step

Solvent StrengthSolvent StrengthSolvent StrengthSolvent Strength

Elution with 20% methanol in water

Elution with 5% methanol in water

Elution with 100% methanol.

Less Retentive SorbentLess Retentive SorbentLess Retentive SorbentLess Retentive Sorbent

Analyte completely eluted from C8

Traces of analyte remain on C18(EC)

Soak StepSoak StepSoak StepSoak Step

The analyte is completely eluted from the column which was soaked for one minute with the elution solvent.

Some analyte remains on the column that was not soaked in the elution solvent.