ENVIRONMENTALENVIRONMENTALANALYSISANALYSISI. Solid Phase ExtractionI. Solid Phase Extraction

ENVIRONMENTALENVIRONMENTALANALYSISANALYSISI. Solid Phase ExtractionI. Solid Phase Extraction

sorbent configurations –

layered phases

Sorbent Configurations Sorbent Configurations to Enhance Selectivityto Enhance SelectivitySorbent Configurations Sorbent Configurations to Enhance Selectivityto Enhance Selectivity

Stacked phases

Layered phases

Mixed phases

Role of Layered PhasesRole of Layered PhasesRole of Layered PhasesRole of Layered Phases

Extending the range

Enhancing the selectivity

Separate cartridges not required

Layered Phases (Loading)Layered Phases (Loading)Layered Phases (Loading)Layered Phases (Loading)

C 2C 2 C 2C 2ENV+ENV+ ENV+ENV+ C 2C 2ENV+ENV+

Small, Small, polarpolar

Large, non-Large, non-polarpolar Broad RangeBroad Range

Layered Phases (Eluting)Layered Phases (Eluting)Layered Phases (Eluting)Layered Phases (Eluting)

ENV+ENV+ C 2C 2 C 2C 2ENV+ENV+

ENV+ENV+

Small, Small, polarpolar

Large, non-Large, non-polarpolar Broad RangeBroad Range

OC Pesticides from WaterOC Pesticides from WaterExtraction on Layered PhasesExtraction on Layered PhasesOC Pesticides from WaterOC Pesticides from WaterExtraction on Layered PhasesExtraction on Layered Phases

Sample pre-treatment

Column solvation

Column equilibration

Sample application

Interference elution

Analyte elution

HCl to pH=2, HCl to pH=2, Methanol, 0.5%Methanol, 0.5%55 mL Methanol mL Methanol

1010 mL reagent water mL reagent water

One liter in One liter in 1/2 1/2 hourhour

10 mL water, 10 mL water, 1010 min N min N22

Two x Two x 22 mL THF mL THF

ISOLUTE® C2 / C18(EC)

OC Pesticides from WaterOC Pesticides from WaterPercent RecoveryPercent RecoveryOC Pesticides from WaterOC Pesticides from WaterPercent RecoveryPercent Recovery

Pesticides C18(EC) C2 C18(EC) SUM

Aldrin 73 85 0 85

4,4’-DDE 62 86 5 91

Endosulfan II 78 65 37 102

AVG

17 pesticides

88 54 43 97

Interference RemovalInterference RemovalInterference RemovalInterference Removal

NH2

Analyte +

Interferences

Analytes (e.g..., PAHs)

ENV+

Interferences (e.g., Humic Acids)

NH2

ENV+

Interferences Retained

AnalytesEluted

AnalytesEluted

Organic Elution Solvent

Extraction of PAHs from Extraction of PAHs from Natural WatersNatural WatersExtraction of PAHs from Extraction of PAHs from Natural WatersNatural Waters

Extraction of analytes such as

PAHs from natural waters, can be

problematic due to co-extracted

humic and fulvic substances that

can interfere with the final analysis

technique.

PAHs and Humic AcidsPAHs and Humic AcidsPAHs and Humic AcidsPAHs and Humic Acids

Naphthalene

Benzo[a]pyrene

[ R - CH - C OO- ]n“Humic Acid”

ISOLUTE PAHISOLUTE PAHISOLUTE PAHISOLUTE PAH

ISOLUTE PAH is designed to alleviate this problem. It consists of a layer of NH2 sorbent above a layer of C18.

The NH2 layer (positively charged at neutral pHs) retains the acidic interferences, while PAHs are retained by and eluted from the C18 layer.

ISOLUTE PAHISOLUTE PAHISOLUTE PAHISOLUTE PAH

NH2

Analyte +

Interferences

Analytes ( PAHs)

C18

Interferences (Humic Acids)

NH2

C18

Interferences Retained

AnalytesEluted

Organic Elution Solvent

SPE ConditionsSPE ConditionsSPE ConditionsSPE Conditions

1. Sample pre-treatment

2. Column solvation

3. Column equilibration

4. Sample application

5. Interference elution

6. Analyte elution

10% IPA

5 mL IPA

5 mL 2% IPA in water

One liter in 30 to 60 min. Bottle wash required.

5 mL water/IPA 9:1 15-30 min drying

2 x 3 mL THF:nC6 (1:1)

ISOLUTE PAH Column (NH2/C18)

(AN 1025)

HPLC ConditionsHPLC ConditionsHPLC ConditionsHPLC Conditions

Column HiChrom PAH2 (150x4.6mmx5m)

Mobile phase A= ACN/Water (30/70, v/v)

B= 100% ACN

Gradient 100% A to 100% B in 35 mins.

100% B for 3 mins

Flow rate 1 ml / min

Detection UV, 254 nm

20 l sample injected

HPLC chromatogram of HPLC chromatogram of blankblank river river water with a high humic acid water with a high humic acid content extracted using content extracted using C18 onlyC18 only

HPLC chromatogram of HPLC chromatogram of blankblank river river water with a high humic acid water with a high humic acid content extracted using content extracted using C18 onlyC18 only

HPLC chromatogram of HPLC chromatogram of blankblank river river water with a high humic acid content water with a high humic acid content extracted using extracted using ISOLUTE PAHISOLUTE PAH

HPLC chromatogram of HPLC chromatogram of blankblank river river water with a high humic acid content water with a high humic acid content extracted using extracted using ISOLUTE PAHISOLUTE PAH

HPLC Chromatogram of 16 PAHs extracted HPLC Chromatogram of 16 PAHs extracted from river water with a high humic acid from river water with a high humic acid content using ISOLUTE PAHcontent using ISOLUTE PAH

HPLC Chromatogram of 16 PAHs extracted HPLC Chromatogram of 16 PAHs extracted from river water with a high humic acid from river water with a high humic acid content using ISOLUTE PAHcontent using ISOLUTE PAH

PAH PAH RecoveriesRecoveriesPAH PAH RecoveriesRecoveries

Naphthalene 62 (13.8)Acenaphthylene 73 (9.5)Acenaphthene 98 (2.1)Fluorene 82 (3.6)Phenanthrene 94 (1.6)Fluoranthene 84 (2.8)Anthracene 101 (2.0) Pyrene 92 (3.1)Benzo(a)anthracene 99 (3.5) Chrysene 100 (3.6)Benzo(b)fluoranthene 94 (4.1)Benzo(k)fluoranthene 95 (4.4)Benzo(a)pyrene 96 (3.5) Indeno(123-cd)pyrene 98 (1.2)Dibenz(ah)anthracene 92 (2.7)Benzo(ghi)perylene 92 (3.1)

Average: 91 (4.0)

ISOLUTE PAH HCISOLUTE PAH HCISOLUTE PAH HCISOLUTE PAH HC

clean-up of HEXANE extracts of

oil contaminated soil

designed to separate the

aromatic PAHs from hydrocarbon

interference (up to C30)

clean baseline in GC analysis

PAH Method (IST AN 1070A)PAH Method (IST AN 1070A)PAH Method (IST AN 1070A)PAH Method (IST AN 1070A)

Action Method

Column conditioning / equilibration

Condition columns with hexane (8ml) at 2ml / min

Sample loading Load 0.5ml of sample in apolar solvent (hexane, i-octane)

Interference elution Rinse column with pentane (3ml) at 0.5ml / min

PAH elution Elute PAHs with 2-propanol (isopropanol) / hexane (3.4 ml in 100ml, v/v), 6ml at 1ml / min

SPE clean-up using ISOLUTE PAH HC, 1g / 3ml

ISOLUTE ISOLUTE EPH ColumnEPH ColumnISOLUTE ISOLUTE EPH ColumnEPH Column

fractionate pentane or hexane extract

of oil contaminated soil

designed to separate the C10-C22

aromatics from C8-C40 aliphatics

proprietary blend of silica / alumina

based on MADEP procedure

PAH Method (IST PAH Method (IST TNTN 1 14242))PAH Method (IST PAH Method (IST TNTN 1 14242))

Action Method

Column conditioning / equilibration

Condition columns with hexane (20ml) under gravity flow

Sample loading Load (max) 1ml of sample in heptane or hexane

Aliphatic elution Elute aliphatics with 12ml of hexane

PAH elution Elute aromatics with 20ml of DCM

SPE clean-up using ISOLUTE EPH-M, 5g / 20ml