Solid Phase Extraction (SPE) - Sorbent Technologies · Solid Phase Extraction (SPE) is a powerful...

84www.sorbtech.com 770-936-0323 Sorbent Technologies, Inc.

Solid Phase Extraction (SPE)Solid Phase Extraction (SPE) is a powerful method for sample preparation and is used by most chromatographers today.

SPE has capabilities in a broad range of applications: • Enviromental analyses • Pharmaceutical and Biochemical analyses • Organic chemistry • Food analysis • Excellent method development tool

The Macherey-Nagel™ and Jordi™ lines of SPE cartridges have the widest diversity of adsorbent types and column sizes.

• Chromabond® SPE Cartridges • Chromabond® QuEChERS

• Chromabond® XTR • Chromabond® - Vacuum Manifold for 12 columns • Jordi™ SPE Cartridges

SPE


SPE


Basic Principles of SPE

In general, SPE is used for three important purposes in state-of-the-art analyses:

• Concentration of the analyte (up to factor 10.000 - increase of chromatographic sensibility and improved limits of detection)• Removal of interfering compounds (protection of subsequent analyses like HPLC, GC, TLC, UV or IR spectroscopy, ...) • Changing an analyte’s environment to a simpler matrix more suitable for subsequent analyses

Advantages of Solid Phase Extraction:• Lower consumption of solvents • Faster-enormous time savings• Less demand placed on analytical equipment• Lower costs per sample• Potential for automation• High consistency in individual sample handling• More specific selectivity because of the broad range of adsorbents and different retention mechanisms• Optimization of extraction by variation or adjusting of the solid phase and chromatographic conditions

Ultra-high performance CHROMABOND® SPE Cartridges are available in polypropylene (standard & Large Volume) and glass tubes from 1 to 70 ml sizes.

Benefits• High recovery rates• Provides variety and selection for even the most specific application• Excellent reproducibility• Improves selectivity of the analysis• All SPE products sealed to maintain quality and shelf life• Reduces consumption of organic solvents• Speeds analysis time• Over 40 Phases available• Cartridges available in polypropylene, glass LV (large volume), 96 well plates

SPE


Main steps of the SPE procedure

1. Conditioning of the adsorbentConditioning of the adsorbent is necessary in order to ensure reproducible interaction with the analyte. Conditioning, also called solvation, results in a wetting of the adsorbent and thus produces an environment which is suitable for adsorption of the analyte. Nonpolar adsorbents are usually conditioned with 2 – 3 column volumes of a solvent, which is miscible with water (methanol, THF, -propanol etc.), followed by the solvent in which the analyte is dissolved (pure matrix, e.g. water, buffer). Polar adsorbents are conditioned with nonpolar solvents. After the conditioning step, the adsorbent bed must not run dry to maintain solvation (deconditioning).

2. Sample application (adsorption)Sample application can be performed with positive or negative pressure with a flow rate of ~3 ml/min. Sample volumes vary from a few milliliters up to liters.

3. Washing of the adsorbentWashing of the adsorbent is usually achieved with a special wash solution; however, in some cases it may not be necessary. If the polarity difference between the wash solution and the eluent is very large, or if both are not miscible, drying of the adsorbent bed after washing is recommended to improve elution and recovery.

4. ElutionElution with a suitable eluent should not be too fast. The elution speed depends on the column or cartridge dimensions and the quantity of adsorbent (about 1 ml/min).

Since analytes can be either adsorbed on the SPE packing material or directly flow through while the interfering substances are retained, two general separation procedures are possible – both cases are shown in the figure below.

Visit www.SORBTECH.com for more information on SPE and Applications!

SPE


Selection GuideMolecular interactions in SPE

SPE adsorbents are most commonly categorized by the nature of their primary interaction mechanism with the analyte of interest. The three most common extraction mechanisms used in SPE are reversed phase (RP), normal phase (NP) and ion exchange.

Typical extraction mechanisms • Reversed Phase Extraction of hydrophobic or polar organic analytes from aqueous matrix • Normal Phase Extraction of polar analytes from non-polar organic solvents • Ion Exchange Extraction of charged analytes from aqueous or non-polar organic samples

Typical extraction mechanismsNonpolar interactions Silica-based: C18 ec, C18, C18 Hydra, C8Polymer-based: HR-X, HR-P, Easy, PS-RPInteraction: hydrophobicSample: mostly aqueousElution: solvents with lower polarity (compared to water) CH3OH, CH2Cl2, CHCl3, hexane

Polar interactionsSilica-based: SiOH, CN, NH2, OH (diol), C6H5Other: Alumina, Florisil®Interaction: hydrogen bonds, dipole-dipole and π-π interactionsSample: mostly organicElution: polar solvents (compared to sample solvent), e.g., (nonprotic) ethers, ketones (MTBE, THF, acetone) CH2Cl2, CHCl3

Cation exchangers Silica-based: SA (SCX), PCA (WCX), PSA,Polymer-based: HR-XC, HR-XCW, PS-H+Interaction: between charged analytes and functional group of cation exchangerSample: aqueous (pH 3-5)Elution: acidic: pH 2 (e.g., HCl, or 20 % AcOH in CH3OH – CH3CN) basic: pH 8-9 (e.g., 5 % NH3 in CH3OH – CH3CN) solvents or buffers with higher ionic strength and counter ions with high selectivity (e.g., Ca2+)

It should be noted, that in SPE the interactions described above are not found in pure form, but in combination. For example, modified silicas, unless they have been subjected to endcapping (silanization of residual silanol groups with short-chain silanes), still possess free silanol groups, which can enter into secondary interactions.

Anion exchangers Silica-based: SB (SAX), NH2, DMAPolymer-based: HR-XA, HR-XAW, PS-OH-Interaction: between charged analytes and functional group of anion exchangerSample: aqueous (pH 8-9)Elution: basic: pH 10 (e.g., 20 % NH3 in CH3OH – CH3CN) acidic: pH 4-5 (e.g., HCl, or 5 % AcOH in CH3OH – CH3CN) solvents or buffers with higher ionic strength and counter ions with high selectivity (e.g., citrate)

SPE


Sample Pretreatment

For direct extraction with adsorbents the sample matrix (sample environment) has to fulfill three conditions: • The matrix has to be liquid, if possible with low viscosity. • Solids should be removed from the liquid matrix. • The matrix (sample environment) should be suit-able for retention of the analyte.

For solid samples there are different methods to convert the sample into a suitable matrix: • Dissolution of the solid sample in a suitable solvent • Lyophilization of the sample and dissolution in a suitable solvent • Extraction of the solid sample with a suitable solvent • Homogenization of the sample in a suitable solvent

In order to find the suitable solvent, one has to consider all desired sample components. Also, the suitable solvent should enhance retention of the analyte. For example, samples with large contents of solids are often homogenized in nonpolar solvents like hexane, while for samples with high water content dissolution in acids, bases, buffers or very polar solvents such as methanol is recommended.

Additionally, SPE allows to alter the properties of the sample matrix. If, for example, natural products are extracted with methanol or acetone, the polarity of the extracts can be increased by dilution with water, in order to enhance nonpolar solid phase extraction on the C18 material.

Design of columns, cartridges, and 96-well plates

CHROMABOND® polypropylene columns• PP columns with PE filter elements• Different sizes from 1, 3, 6 up to 150 mL• Adsorbent weights from 20 mg to 50 g• Male Luer tip as exit• Compatible with most robots (e.g., Gilson ASPEC™,Caliper AutoTrace®)CHROMABOND® glass columns• Glass columns with chemically very inert glass fiber filter elements (nominal pore size 1 μm)• Two different sizes: 3 and 6 mL• Available with all CHROMABOND® phases • Excludes any influence from the column material (e.g., plasticizers)CHROMAFIX® cartridges• PP cartridges with PE filter elements• Three different sizes with different adsorbent weights: Small (0.4 mL), Medium (0.8 mL), Large (1.8 mL) • Female Luer tip at the inlet, male Luer tip as exit• Offers alternative way of handling using positive pressure by syringes or peristaltic pumps• Especially suited for convenient solid phase extraction of small sample volumes

CHROMABOND® LV columns• Large volume PP columns with PE filter elements• Three different adsorbent weights (100, 200 and 500 mg) • Funnel-shaped reservoir with 15 mL volume• Especially for clinical samples - the whole sample (e.g., urine, serum, blood) can be applied to the column in one step• Can be directly used in the Zymate® lab robots of ZymarkCHROMABOND®MULTI 96 SPE in 96-well format• 96-well PP plates with PE filter elements• Cavity volume 1.5 mL• Adsorbent weights 10, 25, 50 and 100 mg• Supplied with any CHROMABOND® SPE adsorbents• For simultaneous preparation of 96 samples• Easy method transfer from CHROMABOND® columns or CHROMAFIX® cartridges to CHROMABOND® MULTI 96• Readily adaptable to all common automated/robotic handling Online-SPE • Online columns and cartridges• SPE columns with caps and needles for the Gerstel MultiPurposeSampler (MPS)• Columns for Gilson ASPEC™ systems (ASP)

SPE


CHROMABOND®- HR-Xpert

The innovative concept of five polymer-based RP- and mixed-mode ion exchange phases for SPE:• CHROMABOND®HR-X - hydrophobic PS/DVB copolymer• CHROMABOND®HR-XC - strong mixed-mode cation exchanger • CHROMABOND®HR-XA - strong mixed-mode anion exchanger• CHROMABOND®HR-XCW - weak mixed-mode cation exchanger • CHROMABOND®HR-XAW - weak mixed-mode anion exchanger

These innovative SPE phases offer:

State-of-the-art spherical polymer• Two particle sizes (45 μm and 85 μm) adequate for different sample volumes and matrices• Broad spectrum of application with special suitability for erichedment of parmaceuticals from biological matrices• Ideal flow properties due to low content of particulate matter

Optimized pore structure and high specific surface• High loadability and outstanding elution properties• Low solvent consumption• Rapid, economical analyses

High - purity adsorber material• Allows highest reproducibility with extremely low blind values• Reliabla analyses at ultra trace level• No method asaptation for new batches necessary

The HR-Xpert concept guarantees:

• RP and mixed-mode SPE phases with distinct ion exchange and reversed phase properties: excellent enrichment of neutral, acidic and basic compounds

• Modern, spherical support polymer with optimized pore structure and high surface: good reproducibility, reliable and cost-efficient analysis

• Possibility for more aggresive washing procedures for matrix removal: cleaner samples and protection of your HPLC and GC instruments

• Quantification of analytes also from heavily contaminated samples: lower limits of detection also for critical matrices

CHROMABOND® HR-Xpert is the perfect combination for all tasks in sample preparationSPE


Chemical structures of the phases:

SPE


2 particle sizes - 1 goal: HR-Xpert for optimized sample preparationFor different application requirements the particle sizes complement each other perfectly

Ideal for:• smaller sample volumes• smaller adsorbtent weights• lower elution volumes

Recommended for:• Large volume or viscous samples, heavy matrix load• operation without vacuum possible (e.g., for volatile analytes)• higher adsorbent weight without increase in back pressure

Features of 45 µm particles• About half the radius results in 8fold particle number per volume for approx. equal adsorbent weight• Same specific surface for both particle sizes: considerably larger freely accessible external surface for 45 μm particles• Denser adsorbent packing: enhanced interaction of the analyte with the adsorbent, better extraction results.

Ideal elution characteristicsMethod: 1mL column with 30 mg CHROMABOND® HR-X, 1 mL standard solution (1mg/mL hexobarbital), drying, elution in portions of 100 μL with methanol.

Advantages of 45 µmparticles:• faster elution• lower elution volumes required

Breakthrough behavior in enrichmentMethod: 1mL column with 15 mg CHROMABOND® HR-X, apply portions of 1 mL standard solution (250 μg/mL hexobarbital in water), collect eluates.

45 µm (red)the analyte is completely retained up to fraction 10.

85 µm (blue)Small amounts even break through with fraction 4.45 μm particles provide better enrichment and breakthrough behavior for small adsorbent weights.When using larger adsorbent weights this effect isless pronounced, since then analytes have sufficient contact with the 85 μm adsorbent particles as well.

45 µm particles are ideal for small sample and elution volumes, while for large amounts of sample and adsorbent 85 µm particles show advantages due to better flow properties.

SPE


3 paths – 1 goal: cleaner samplesDepending on the character of the analytes HR-Xpert offers suitable adsorbents and

optimal method for sample preparation, cleaning and concentration.

Strong Bases Strong AcidsAcidic

CompoundsNeutral

CompoundsBasic

Compounds

strong basespKa > 10

CHROMABONDHR-XCW

acidspKa 2 > 8

CHROMABONDHR-XA

neutralscompounds

CHROMABONDHR-X

strong acidspKa > 1 CHROMABOND

HR-XAW

basespKa 2 -10

CHROMABONDHR-XC

CHROMABOND HR-Xpert concept neutral, acidic, and basic

analytes

Method A1. Sample pretreatment2. Column conditioning3. Sample application4. Washing 1: (basic) aqueous5. Washing 2/ Elution 1*:

organic6. Elution 2: organic acidic

Method C1. Sample pretreatment2. Column conditioning3. Sample application4. Washing 1: (acidic) aqueous5. Washing 2/ Elution 1*:

organic6. Elution 2: organic base

Method B1. Sample pretreatment2. Column conditioning3. Sample application4. Washing 1: aqueous5. Elution: organic

SPE


HR-X Properties: • Hydrophobic polystyrene-divinylbenzene copolymer• pH stability 1 – 14• High-purity material with highest reproducibility and lowest blank values due to a novel manufacturing process• Spherical particles 85 μm; pore size 55 – 60 Å• Very high surface 1000 m2/g• Excellent recovery rates especially for enrichment of pharmaceuticals

• Capacity 390 mg/g (caffeine in water)

Recommended Applications:• Pharmaceuticals / active ingredients from tablets, creams and water / waste water• Drugs and pharmaceuticals from urine, blood, serum and plasma• Trace analysis of pesticides, herbicides, phenols, PAHs and PCBs from water

Cat. # Product Weight Size Qty/pkg730934 Chromabond®, HR-X 30 mg 1 ml 30730931 Chromabond®, HR-X 200 mg 3 ml 30730936 Chromabond®, HR-X 60 mg 3 ml 30730936G Chromabond®, HR-X - Glass 60 mg 3 ml 30730937 Chromabond®, HR-X 500mg 3 ml 30730938 Chromabond®, HR-X 200 mg 6 ml 30730939 Chromabond®, HR-X 500mg 6ml 30732130 Chromabond® LV, HR-X - Large Volume 30 mg 15 ml 30732131 Chromabond® LV, HR-X - Large Volume 60 mg 15 ml 30

SPE

For more selections and sizes, visit our website or contact your personal Product Specialist.

Drugs from WaterColumn type:CHROMABOND® HR-X, 3 mL, 200mg

Sample: 1μg.mL each in waterColumn conditioning: 5 mL methanol, 5 mL dist. waterSample application: slowly aspirate 500 mL water (pH3) throughthe columnColumn washing: 5 mL waterElution: after drying 3 x 2 mL acetonitrileFurther analysis: HPLC on NUCLEODUR® C18 Gravity, 5 μm

Recovery rate [%]

Compound HR-X Strata™ XKetoprofen 98 92Ibuprofen 91 93Pentobarbital 99 95Meclofenamic Acid 92 93Protriptyline 63 45Nortriptyline 53 39

Pesticides from WaterColumn type:CHROMABOND® HR-X, 3 mL, 200mg

Sample: samples are spiked with 500 ng of each pesticidein 1000 nL water, adjusted to pH 2 with HCI or pH 7Column conditioning: 10 mL methanol, 10 mL dist. waterSample application: Slowly pass 1000 mL spiked water sample through the column with the aid of a tubing adapterElution: after drying 5 mL methanol - THF (1:1, v/v)Further analysis: HPLC

Recovery rate [%]

Compound HR-X Compound HR-X pH 2 pH 7

Metamitron 86 Desisopropylatrazine 90Quinmerac 90 2, 4-Dichlorobenzamide 95Chloridazon 93 Desethylatrazine 89Picloram 83 Hexazinone 95Metribuzin 84 Bromacil 103Cyanazine 83 Simazine 91Metabenzthiazuron 94 Desethylterbuthylazine 89Chlortoluron 91 Atrazine 88Isoproturon 89 Metalaxyl 97Diuron 91 Metazachlor 93Dimethenamid-P 89 Propazine 88Linuron 94 Terbuthylazine 86Epoxyconazole 85 Metolachlor 97Penconazole 90Alachlor 93Propiconazole-1 89Flufenacet 91Diflufenicam 58Triallate 42

Standard protocol for CHROMABOND® HR-XColumn type:CHROMABOND® HR-X, 3 mL, 200 mg

Sample pretreatment: if necessary, adjust pH valueColumn conditioningL 5 mL methanolEquilibration: 5 mL waterSample application: Slowly aspirate the sample through the columnColumn washing: 5 mL water - methanol (95:5, v/v)Elution: after drying 3 x 2 mL methanolFurther analysis: if necessary, evaporate and redissolve in a suitablesolvent; HPLC or GC


HR-XC Properties: • Base material polystyrene-divinylbenzene copolymer• Strong acidic benzenesulphonic acid cation exchanger • Exchange capacity 1.0 meq/g, pKa < 1• pH stability 1 – 14• High purity material, highest reproducibility and lowest blank values due to an optimized production process• Spherical particles size 85 μm; pore size 65 – 75 Å• Very large specific surface 800 m2/g; pore volume 1.4 cm3/g

• RP capacity 300 mg/g (caffeine in water)• Outstanding recovery rates especially for the enrichment of basic analytes.

Recommended Applications:• Basic active ingredients from heavily matrix-contaminated samples, e.g. urine, plasma, serum• Fungicides from food, melamine from milk basic analytes, e. g. amines• Bases with pKa 2 – 10

Cat. # Product Weight Size Qty/pkg730969 Chromabond®, HR-XC 30 mg 1 ml 30730956 Chromabond®, HR-XC 60 mg 3 ml 30730049 Chromabond®, HR-XC 100 mg 1 ml 30730957 Chromabond®, HR-XC 150 mg 6 ml 30730952 Chromabond®, HR-XC 200 mg 3 ml 30730953 Chromabond®, HR-XC 500 mg 3 ml 30730955 Chromabond®, HR-XC 500 mg 6 ml 30731755 Chromafix®, HR-XC 155 mg S 50731756 Chromafix®, HR-XC 240 mg M 50

Standard protocol for CHROMABOND® HR-XCColumn type:CHROMABOND® HR-XC, 3 mL, 200 mg

Sample pretreatment: individual sample preparation with reference to analytes and matrixColumn conditioning: 5mL methanolEquilibration: 5 mL waterSample application: Slowly aspirate the sample through the columnWashing 1: 2 mL 0.1 mol/L HCI in waterWashing 2/Elution 1: 2 mL methanol (neutral and acidic compounds); if necessary, further washing stepsElution 2: after drying 5mL methanol - 5% NH3 (basic compounds)Further analysis: if necessary, evaporate and redissolve in a suitablesolvent; HPLC or GC

Fractionation of acidic, neutral and basic analytes from serumColumn type:CHROMABOND® HR-XC, 3 mL, 200mg

Sample: 1 mL spiked matrix, acidified with 200 μL 2% h3PO4Column conditioning: 5 mL methanol, then 5 mL dist. waterSample application: slowly aspirate sample through the columnWashing: 2 mL 0.1 mol/L HCIElution: 2.5 ml methanol (fraction A: neutral and acidic analytes);then 5 mL methanol - NH3 90:10, v/v (fraction B: basic analytes)

Recovery rate [%]Fraction A: Fraction B:neutral and acidic basic analytesanalytes

Compound HR-XC Compound HRXC Oasis® Strata™ MCX X-C Suprofen 108 1.Doxepin 101 68 82Naproxen 85 2.Imipramine 95 71 85Tolmetin 73 3.Amitriptyline 94 72 78Phenobarbital 108 4.Trimipramine 92 70 81Indomethacin 33 Hexobarbital 80

SPE



HR-XAProperties:• Base material polystyrene-divinylbenzene copolymer• Strong basic quaternary ammonium anion exchanger • Exchange capacity 0.25 meq/g, pKa ~ 18• pH stability 1 – 14• High purity material with highest reproducibility and lowest blank values due to an optimized production process• Spherical particles size 85 μm; pore size 55 – 65 Å• Very large specific surface 850 m2/g; pore volume 1.4 cm3/g• RP capacity 350 mg/g (caffeine in water)

• Outstanding recovery rates especially for the enrichment of acidic analytes

Recommended Applications: • Acidic active ingredients from heavily matrix-contaminated samples like e. g. urine, plasma, serum• Phenolic acids• Acidic herbicides• Weak/medium-strength• Acids with pKa 2 – 8

Cat. # Product Weight Size Qty/pkg730968 Chromabond®, HR-XA 30 mg 1 ml 30730950 Chromabond®, HR-XA 60 mg 3 ml 30730727 Chromabond®, HR-XA 100 mg 1 ml 30730951 Chromabond®, HR-XA 200 mg 3 ml 30730958 Chromabond®, HR-XA 150 mg 6 ml 30730954 Chromabond®, HR-XA 500 mg 3 ml 30730966 Chromabond®, HR-XA 500 mg 6 ml 30731768 Chromafix®, HR-XA 155 mg - 50731769 Chromafix®, HR-XA 240 mg - 50

Standard protocol for CHROMABOND® HR-XA

Column type: CHROMABOND® HR-XA, 3 mL, 200 mgSample pretreatment: individual sample preparation with referenceto analytes and matrixConditioning: 5 mL methanolEquilibration: 5 mL waterSample application: Slowly aspirate the sample through the columnWashing 1: 2 mL 0.1 mol/L NaOH in waterWashing 2/Elution 1: 2 mL methanol (neutral and basic compounds), if necessary, further washing stepsElution 2: after drying 5 mL methanol - 1 to 10% formic acid(acidic compounds)Further analysis: if necessary, evaporate and redissolve in a suitablesolvent; HPLC or GC

Acidic Herbicides on CHROMABOND® HR-XA

Column type: CHROMABOND® HR-XA, 3 mL, 200 mgSample pretreatment: 1 μg/mL, adjusted on basic pH with 1 N NaOHConditioning: 5 mL methanol, then 5 mL water(Do not let run the column dry!)Sample aspiration: The prepared sample is passed through the column by vacuumWashing: With the following washing mixtures impurities are removed: a) 2.5 mL 50 mM NaOAc b) 2.5 mL methanolDrying: With nitrogen or airElution: Analytes are eluted with 5 mL methanol / 10 % formic acid

Evaporation to dryness and reconstitution with 1 mL of mobile phase from subsequent HPLC

Subsequent analysis: HPLC determination of recovery rates withEC 125/4 NUCLEODUR® C18 Gravity, 5 μm (REF 760100.40) in reference to Appl. # 122820

Phenolic Acids on CHROMABOND® HR-XA

Column type: CHROMABOND® HR-XA, 3 mL, 200 mgSample pretreatment: 1 μg/ml, adjusted on basic pH with 1 N NaOHConditioning: 5 ml methanol, then 5 ml water (Do not let run the column dry!)Sample aspiration: The prepared sample is passed through the column by vacuumWashing: With the following washing mixtures impurities are removed: a) 2.5 ml water b) 2.5 ml 0.1 N NaOH c) 5.0 ml methanolDrying: With nitrogen or airElution: Analytes are eluted with 5 ml methanol / 5 % formic acid

Evaporation to dryness and reconstitution with 1 ml of mobile phase from subsequent HPLC

Subsequent analysis: HPLC determination of recovery rates with EC 125/4 NUCLEODUR® 100-5 C18 ec (Cat. No. 760001.40) in reference to Appl. No 122830

HR-XA Oasis MaxDicamba 87 % 78 %Bentazon 79 % 72 %

2,4-D 82 % 80 %MCPA 76 % 72 %

HR-XA Oasis Maxp-hydroxybenzoic acid 97 % 97 %

vanillinic acid 86 % 85 %syringic acid 70 % 60 %

SPE



Cat. # Product Weight Size Qty/pkg730731 Chromabond®, HR-XCW 30 mg 1 ml 30730735 Chromabond®, HR-XCW 60 mg 3 ml 30730733 Chromabond®, HR-XCW 100 mg 1 ml 30730737 Chromabond®, HR-XCW 150 mg 6 ml 30730739 Chromabond®, HR-XCW 200 mg 3 ml 30730741 Chromabond®, HR-XCW 500 mg 3 ml 30730743 Chromabond®, HR-XCW 500 mg 6 ml 30731774 Chromafix®, HR-XCW 155 mg - 50731775 Chromafix®, HR-XCW 240 mg - 50

HR-XCWProperties: • Base material spherical PS/DVB copolymer• Weak acidic carboxylic acid cation exchanger• Exchange capacity >0.7 meq/g, pKa ~ 5• pH stability 1 – 14• High purity material, highest reproducibility and lowest blank values due to an optimized production process• Spherical particles size 85 μm; pore size 50 – 60 Å• Very large specific surface 850 m2/g; pore volume 1.2 – 1.4 cm3/g• RP capacity 350 mg/g (caffeine in water)• Outstanding recovery rates especially for enrichment of strongly basic analytesRecommended Applications: • Basic compounds like• Quaternary amines• Active ingredients from heavily matrix-contaminated, samples e.g. urine, plasma, serum• Strong bases with pKa > 10

Compound HR-XCW

HR-XC*

PCA** Oasis® WCX

Doxepine 79 5 11 41Imipramine 79 9 20 67Amitriptyline 91 9 14 46Trimipramine 98 7 14 27

Tricyclic AntidepressantsAppl. No. 305340Column type: CHROMABOND® HR-XCW/3 ml, 60 mg, REF 730735Sample: 250 μl spiked Serum, diluted with 1 ml 10%, formic acid in waterConditioning: 3 ml MeOHEquilibration: 3 ml waterSample app.: slowly aspirate sample through the columnWashing: 1 ml 5% formic acid in water, then 1 ml MeOHElution: After drying by vacuum (15 min) 3 ml 5% formic acid in MeOHFurther analysis: Evaporate and redissolve in a suitable solvent for HPLC on NUCLEODUR® C8, Gravity see Appl. No. 118520

Recovery rate [%]:

* HR-XC: Basic analytes can not be eluted with slightly acidic organic conditions from the strong cation exchanger CHROMABOND® HR-XC, because the eluting power is not sufficient to dissociate the interaction with the ion exchanger. However, with the usage of basic methanol a complete elution can be achieved (please see also Appl. No. 304780).** PCA: Due to the missing RP interactions of silica based weak cation exchanger, CHROMABOND® PCA gives only a small enrichment elution of the analytes.

Standard protocol for CHROMABOND® HR-XCWColumn type:CHROMABOND® HR-XCW, 3 mL, 200 mg

Sample pretreatment: individual sample preparation with referenceto analytes and matrixConditioning: 5 mL methanolEquilibration: 5 mL acidified waterSample application: Slowly aspirate the sample through the columnWashing 1: 2 mL acidified waterWashing 2/Elution 1: 2 mL methanol (neutral and basic compounds), if necessary, further washing stepsElution 2: after drying 2x2 mL methanol - 1 to 5% formic acid(strongly basic compounds)Further analysis: if necessary, evaporate and redissolve in a suitablesolvent; HPLC or GC

SPE



Cat. # Product Weight Size Qty/pkg730728 Chromabond®, HR-XAW 30 mg 1 ml 30730747 Chromabond®, HR-XAW 60 mg 3 ml 30730729 Chromabond®, HR-XAW 100 mg 1 ml 30730748 Chromabond®, HR-XAW 200 mg 3 ml 30730749 Chromabond®, HR-XAW 150 mg 6 ml 30730744 Chromabond®, HR-XAW 500 mg 3 ml 30730745 Chromabond®, HR-XAW 500 mg 6 ml 30731771 Chromafix®, HR-XAW 155 mg - 50731772 Chromafix®, HR-XAW 240 mg - 50

HR-XAW Properties:• Base material spherical PS/DVB copolymer, pH stability 1 – 14• Weak basic secondary and tertiary ammonium anion exchanger, exchange capacity >0.5 meq/g, pKa ~ 6• High purity material with highest reproducibility and lowest blank values due to an optimized production process• Spherical particles size 85 μm; pore size 55 – 65 Å• Very large specific surface 850 m2/g; pore volume 1.2 – 1.4 cm3/g RP capacity 350 mg/g (caffeine in water)• Outstanding recovery rates especially for enrichment of acidic analytes

Recommended Applications: • Perflourinated surfactants• Acidic compunds like sulfonates• Active ingredients from heavily matrix-contaminated, samples e.g. urine, plasma, serum• Strong acids with pKa < 1

Standard protocol for CHROMABOND® HR-XAWColumn type:CHROMABOND® HR-XAW, 3 mL, 200 mg

Sample pretreatment: individual sample preparation with referenceto analytes and matrixConditioning: 5 mL methanolEquilibration: 5 mL waterSample application: Slowly aspirate the sample through the columnWashing 1: 25 mmol/L ammonium acetateWashing 2/Elution 1: 2 mL methanol (neutral and basic compounds), if necessary, further washing stepsElution 2: after drying 2 x 2 mL methanol - 1 to 5% ammonia(strongly acidic compounds)Further analysis: if necessary, evaporate and redissolve in a suitablesolvent; HPLC or GC

Analysis of perfluorinated surfactants from waterColumn: 125 x 2 mm NUCLEODUR® Sphinx RP, 3 μm Application in accordance with DIN 38407-42Eluent: A) 10 mmol/L NH4Ac in water - methanol (75:25, v/v) Column type: B) 10 mmol/L NH4Ac in acetonitrile - methanol (75:25, v/v) CHROMABOND® HR-XAW, 3 mL, 60 mg 10-30% B in 3 min, 30-55% B in 8 min. Sample: 500 mL water, spiked with 1 mL standard solution 55-10% B in 4 min (20 μg/L of each compound)Flow rate: 0.30 mL/min, temperature 50 ºC Conditioning: 2 mL methanol + 5% ammonia, then 2 mL Injection: 2.5 μL (5 mg/L each after SPE enrichment) methanol, finally 2 mL water.Detection: MS, ESI negative Sample application: slowly aspirate sample through the column. Washing: 2 mL water, then 2 mL acetone - acetonitrile - formic acid (50:50:1 v/v/v), finally 2 mL methanol Elution: 2 mL methanol with 5% ammonia Further analysis: evaporate to dryness in a stream of nitrogen under slight heating, and redissolve in a suitable solvent for HPLC Recovery rates [%]: Compound Recovery 1. Perfluoropropionic acid (PFPrA) 103 2. Perflouropentanoic acid (PFPeA) 94 3. Perflourohexanoic acid (PFPHxA) 95 4. Perflourooctanoic acid (PFOA) 95 5. Perfluorooctane sulfonate K salt (PFOS) 81 6. Perfluorododecanoic acid (PFDoDA) 82

SPE



Easy - Polar, bifunctionally modified polystyrene-divinylbenzene copolymerBecause of the bifunctional modification, CHROMABOND® Easy is significantly more hydrophilic than conventional polystyrene-divinylbenzene polymers and is therefore easily wettable with water. Aqueous sample can be loaded directly with little or no preconditioning. There is no decrease in recovery rates if the cartridge runs dry and therefore automation is easier because a permanent vacuum can be used without supervision.

Properties:• Polar modified polystyrene-divinyllbenzene copolymer with a weak anion exchanger• Specific surface area 650 - 700 m2/g, particle size 80 μm, pore size 50 Å, pH stability 1 - 14• Without preconditioning• Due to bi functional modifcation much more hydrophilic than conventional polystyrene-divinyllbenzene polymers• Easily wettable with water

Cat. # Product Weight Size Qty/pkg730753 Chromabond®, Easy 60 mg 3 ml 30730754 Chromabond®, Easy 200 mg 3 ml 30730756 Chromabond®, Easy 500 mg 6 ml 30730753G Chromabond®, Easy 60 mg 3 ml 30730755G Chromabond®, Easy - Glass 200 mg 6 ml 30

Recommended Applications:• Polar herbicides / pesticides from water (acidic, neutral, basic)• Polar phenols from water• Polyaromatic compounds• Polychlorinated biphenyls• Drug analysis from urine, blood, serum, plasma, pharmaceuticals / active ingredients from tablets, creams


Recovery rates of Phenyl urea pesticides

Cartridge: CHROMABOND®; EASY 3 ml 200 mg

Phase: CHROMABOND Easy

Catalog #: 730754

Substances: fluometuron fenuron monuron isoproturon chlortoluron linuron diuron metoxuron metobromuron monolinuron

Information: Drinking water; internal standard 100 mg/l of each substance

Conditions:

• Conditioning: 2 ml methanol, 2 ml water Sample • Application: Suck 1 L water slowly through the column and

dry with vacuum Elution: 2 x 2 ml methanol/acetone 1:1, then evaporate at 20° C. Legend: Substance Recovery rate % Fluometuron 93 Fenuron 83

Monuron 91 Isoproturon 91 Chlortoluron 95 Linuron 106 Diuron 99 Metoxuron 86 Metobromu

Recovery of PesticidesPrivate communication : Mr. Kühn, GUB, Waldshut Tiengen, Germany

Column type: CHROMABOND® Easy/ 3 ml/ 200 mgColumn conditioning: 1 ml water, 3 ml methanol, 1 ml waterSample application: aspire the sample through the columnElution: 3 x 1 ml acetoneFurther analysis: HPLC with NUCLEOSIL® 120-5 C18

Compound Recovery Compound RecoveryDesisopropylatrazine 90.3% Matalaxyl 95.5%2,6-Dichlorobenzamide 93.1% Isoproturon 93.5%Desethylatrazine 92.7% Diuron 94.4%Hexazinone 69.3% Metazachlor 97.0%Terbacil 65.1% Propazine 94.6%Simazine 81.4% Terbuthylazine 93.2%Cyanazine 92.8% Linuron 95.7%Desethylterbuthylazine 90.6% Metolachlor 97.3%Methabenzthiazuron 93.7% Triallate 61.4%Chlortoluron 91.4% Standard 63.7%Atrazine 92.4%

SPE


HR-PProperties:• Highly porous polystyrene-divinylbenzene copolymer Specific surface 1200m2/g particle size 50-100 μm• Very high binding capacity, up to 30% od adsorbent weight (for comparison: silica adsorbents about 3%

Recommended application:• aromatic compounds• phenols from water• nitroaromatics from water • PAHS from oil

Cat. # Product Weight Size Qty/pkg730108 Chromabond®, HR-P 200mg 3ml 30730117 Chromabond®, HR-P 500mg 3ml 30730111 Chromabond®, HR-P 500mg 6ml 30

PS-RP / PS-OH- / PS-H+ / PS-Mix / PS-Ag+ / PS-Ba2+Properties:• Base material high purity polystyrene-divinylbenzene copolymers (PS/DVB), pore size 100Å, particle size 100μm.• Very low degree of swelling, thus very well suited for Chromatography.• Reliable function over the whole pH range from 0-14 Different modifications for different applications from elimination of nonpolar compounds up to the removal of specific polar components.

Recommended application:Removal of interfering compounds• Improves chromatographic separation, if the interfering components oveerlap with the analyte in the chromatogram• Improves lifetime of the chromatographic column, since interfering components can irreversibly block the column packing• Enrichment of the analytes

Cat. # Product Weight Size Qty/pkg730765 Chromabond®, PS-RP 200mg 3ml 30730692 Chromabond®, PS-RP 500mg 3ml 30730693 Chromabond®, PS-RP 500mg 6ml 30730396 Chromabond®, PS-OH- 200mg 3ml 30730344 Chromabond®, PS-OH- 500 mg 3 ml 30730378 Chromabond®, PS-OH- 500 mg 6ml 30730690 Chromabond®, PS-H+ 200mg 3ml 30

730376 Chromabond®, PS-H+ 500 mg 3 ml 30730377 Chromabond®, PS-H+ 500mg 6ml 30

Aromatic amines from water samplesCompounds investigated: aromatic amines Sample application: aspirate sample through the column with about 10 mL/Column type: ColumnWashing: wash with 3 mL dist. water, dry 5 min under CHROMABOND® HR-P, 3 mL, 200 mg vacuum Elution: 3 x 1 mL methanol - acetonitrile (1:1, v/v) Sample pretreatment: adjust to pH 9 using 10 mol/L NaOHColumn Conditioning: 2 mL each of methanol, acetonitrile and10-5 mol/L sodium hydroxide

Application 301930/302750: removal of halides from aqueous samples shown for the trace analysis of nitrate besides an excess of chloride or bromide

Compounds investigated: 20 ppm nitrate besides 2500 ppm chloride Sample application and elution:or 500 ppm bromide, respectively apply 4 x 1 mL sample factions to the cartridge, discards 1st mL, collect 2nd, 3rd and 4th mLColumn type: separatelyCHROMAFIX® PS-Ag+ (M) Further analysis: HPLC with column 250 x 4 mm0.8 mL, Ø 480 mg NUCLEOSIL® Anion II; eluent 2 mmol/L K HColumn conditioning: 1 mL dist. water phthalate pH 6, 2 mL/min; detection: indirect UV, 280 nm

SPE


Recommended Applications:• Aflatoxins• Chloramphenicol • Pesticides • Steroids • Vitamins

Silica GelProperties:• Unmodified, weakly acidic silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Very polar• Adsorbs humidity from air, for this reason it should be kept well closed and if necessary dried before use• Due to its high affinity for polar compounds it should not be conditioned with polar (e.g. methanol) or water-containing solvents

Cat. # Product Weight Size Qty/pkg730070 Chromabond®, Silica 500 mg 6 ml 30730075 Chromabond®, Silica 1 g 6 ml 30730217 Chromabond®, Silica 2 g 15 ml 20730072 Chromabond®, Silica 10 g 70 ml 10730214G Chromabond®, Silica - Glass 200 mg 3 ml 50730075G Chromabond®, Silica - Glass 1 g 6 ml 30

C18 ecProperties:• Base material silica, pore size 60 Å, particle size 45 μm for C18 ec, specific surface 500 m2/g, pH stability 2 – 8• Octadecyl phases, end-capped, carbon content 14%• Very nonpolar, hydrophobic interactions with a wide variety of organic compounds• Advantageous for clean-up of samples with large structural variations (polarity differences)

Recommended Applications:• Nonpolar compounds• Aflatoxins, amphetamines, antibiotics, Anti-epileptics, barbiturates, caffeine, drugs, preservatives, fatty acids, nicotine, PAHs, pesticides, PCBs, heavy metals, vitamins• Very well suited for desalting of samples

Cat. # Product Weight Size Qty/pkg730011 Chromabond®, C18 ec 100 mg 1 ml 100730012 Chromabond®, C18 ec 200 mg 3 ml 50730013 Chromabond®, C18 ec 500 mg 3 ml 50730014 Chromabond®, C18 ec 500 mg 6 ml 30730015 Chromabond®, C18 ec 1 g 6 ml 30730404 Chromabond®, C18 ec 2 g 15 ml 20730405 Chromabond®, C18 ec 5 g 45 ml 20730259 Chromabond®, C18 ec 10 g 70 ml 10730012G Chromabond®, C18 ec - Glass 200 mg 3 ml 50730014G Chromabond®, C18 ec - Glass 500 mg 6 ml 30732012 Chromabond® LV, C18 ec - Large Volume 200 mg 15 ml 30732013 Chromabond® LV, C18 ec - Large Volume 500 mg 15 ml 30

Anti-epileptics from serumCartridge: CHROMABOND®; C18 ec / 1 ml / 100 mgPhase: CHROMABOND C18 ecCatalog #: 730011

Substances: Carbamazepine phenobarbital phenytoin primidone Information: Mix 500 μl serum with 500 μl internal standard (500 μg / ml 4-methylprimidone in citrate buffer pH

Conditions: Column conditioning: 2 column volumes methanol, 2 column volumes water Sample application: Slowly force or aspirate the sample through the column Column washing: 2 column volumes dist. water.

SPE



Recommended Applications:• Pesticides, PCBs

C8Properties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Octyl phases, not end-capped, carbon content 8%• Similar to C18, however slightly more polar• Secondary interactions with polar compounds are more pronounced due to shorter alkyl chains

Recommended Applications:• Compounds, which are too strongly retained on C18 or C8 e.g. analgetics from blood

C4Properties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Butyl phase, not end-capped, carbon content 7%• Slightly more polar than C18 or C8• Due to shorter alkyl chains the silica surface is not completely shielded

Cat. # Product Weight Size Qty/pkg730021 Chromabond®, C8 100 mg 1 ml 100730022 Chromabond®, C8 200 mg 3 ml 50730023 Chromabond®, C8 500 mg 3 ml 50730024 Chromabond®, C8 500 mg 6 ml 30730134 Chromabond®, C8 1000 mg 6 ml 30730024G Chromabond®, C8 - Glass 500 mg 6 ml 30732023 Chromabond® LV, C8 - Large Volume 500 mg 15 ml 30

Cat. # Product Weight Size Qty/pkg730225 Chromabond®, C4 100 mg 1 ml 100730227 Chromabond®, C4 500 mg 3 ml 50

Recommended Applications:• E.g. anti-epileptics from plasma

C2Properties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Dimethyl phase, not end-capped, carbon content 4 %• Similar to C4

Cat. # Product Weight Size Qty/pkg730169 Chromabond®, C2 100 mg 1 ml 100730221 Chromabond®, C2 500 mg 3 ml 50730409 Chromabond®, C2 500 mg 6 ml 30730410 Chromabond®, C2 1 g 6 ml 30

SPE


Recommended Applications:• Aflatoxins• Caffeine • Phenols

PhenylProperties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Phenyl phase, carbon content 8% • Polarity similar to C8• In addition to hydrophobic interactions, more selective adsorption is possible by π-π interactions due to the electron density of the phenyl ring

Recommended Applications:• Trace elements • Lipids

Amino (NH2)Properties:• Base material silica, pore size 60 Å, particle size 45, specific surface 500 m2/g, pH stability 2 – 8• Aminopropyl phase, carbon content 3.5% • Polar, weak anion exchanger

Cat. # Product Weight Size Qty/pkg730083 Chromabond®, Phenyl (C6H5) 100 mg 1 ml 100730411 Chromabond®, Phenyl (C6H5) 200 mg 3 ml 50730084 Chromabond®, Phenyl (C6H5) 500 mg 3 ml 50

Cat. # Product Weight Size Qty/pkg730031 Chromabond®, Amino (NH2) 100 mg 1 ml 100730413 Chromabond®, Amino (NH2) 200 mg 3 ml 50730033 Chromabond®, Amino (NH2) 500 mg 3 ml 50730180 Chromabond®, Amino (NH2) 500 mg 6 ml 30730626 Chromabond®, Amino (NH2) 1 g 6 ml 30730033G Chromabond®, Amino (NH2) - Glass 500 mg 3 ml 50730180G Chromabond®, Amino (NH2) - Glass 500 mg 6 ml 30730626G Chromabond®, Amino (NH2) - Glass 1 g 6 ml 30732033 Chromabond® LV, Amino (NH2) - Large Volume 500 mg 15 ml 30

Recommended Applications:• Together with phase SA for PCB and pesticides

AluminaProperties:• Aluminium oxide, high purity, pore volume 0.90 ml/g, particle size 60 – 150 μm, specific surface 150 m2/g

Cat. # Product Weight Size Qty/pkg730452 Chromabond®, Alumina Acid 500 mg 3 ml 50730429 Chromabond®, Alumina Basic 500 mg 3 ml 50730446 Chromabond®, Alumina Neutral 500 mg 3 ml 50730017 Chromabond®, Alumina Acid 1 g 6 ml 30730139 Chromabond®, Alumina Neutral 1 g 6 ml 30730467 Chromabond®, Alumina Basic 4 g 45 ml 20730139G Chromabond®, Alumina Neutral - Glass 1 g 6 ml 30730020G Chromabond®, Alumina Basic - Glass 1 g 6 ml 30732210 Chromabond® LV, Alumina Acid - Large Volume 1 g 15 ml 30732091 Chromabond® LV, Alumina Neutral - Large Volume 1 g 15 ml 30732205 Chromabond® LV, Alumina Basic - Large Volume 1 g 15 ml 30

SPE


Recommended Applications:• Cyclosporins• Carbohydrates

CyanoProperties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Cyanopropyl phase, carbon content 5.5% • Polar to mid-polar• In addition to weak hydrophobic interactions, selective interactions are possible due to the high electron density of the Cyano group

Cat. # Product Weight Size Qty/pkg730061 Chromabond®,Cyano 100 mg 1 ml 100730420 Chromabond®, Cyano 200 mg 3 ml 50730063 Chromabond®, Cyano 500 mg 3 ml 50730421 Chromabond®, Cyano 500 mg 6 ml 30

Recommended Applications:• Antibiotics • Prostaglandins

DiolProperties:• Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2 – 8• Diol phase, carbon content 5.5%• Polar • Properties similar to SiOH

Cat. # Product Weight Size Qty/pkg730051 Chromabond®, Diol 100 mg 1 ml 100730417 Chromabond®, Diol 200 mg 3 ml 50730053 Chromabond®, Diol 500 mg 3 ml 50730418 Chromabond®, Diol 500 mg 6 ml 30730053G Chromabond®, Diol - Glass 500 mg 3 ml 50732053 Chromabond® LV, Diol - Lg. Vol. 500 mg 15 ml 30

Also available in Glass!Florisil®Properties:• Matrix magnesium silicate (MgO - SiOH 15:85)• High purity• Particle size 150 – 250 μm

Recommended Applications:• Organic tin compounds, aliphatic carboxylic acids, PCBs, PAHs


Cat. # Product Weight Size Qty/pkg730457 Chromabond®, Florisil® 200 mg 3 ml 50730081 Chromabond®, Florisil® 500 mg 3 ml 50730238 Chromabond®, Florisil® 500 mg 6 ml 30730082 Chromabond®, Florisil® 1 g 6 ml 30730082G Chromabond®, Florisil® - Glass 1 g 6 ml 30

Carbon AProperties: • Base material: activated carbon, highly porous, spherical particles, specific surface >1000 m2/g

Recommended application:• Recommended application:• Determination of acrylamide from water according to DIN 38413-6• Note:Acrylamide and the isotopically labeled form are carcinogenic, mutagenic and neurotoxic.

Cat. # Product Weight Size Qty/pkg730165 Chromabond®, Carbon A 500 mg 6 ml 30730167 Chromabond®, Carbon A 1000 mg 6 ml 30

SPE


SA (Strongly acidic cation exchanger)Properties:• Base material silica, pore size 60Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2- 8• Benzenesulfonic acid modified silica strongly acidic cation exchanger (capacity ~ 0.5 meq/g)• Ion exchange of organic compounds from aqueous matrix elution of interesting compounds with solvent systems, which compensate the ionic and nonpolar interactions, e.g., methanolic HCI

Recommended application:• amino acids• amines• chlorophyll• PCB

Cat. # Product Weight Size Qty/pkg730322 Chromabond®, SB 200mg 3ml 50730079 Chromabond®, SB 500mg 3ml 50730426 Chromabond®, SB 500mg 6ml 50

SB (Strongly basic anion exchanger)Properties:• Base material silica, pore size 60Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2- 8• Silica modified with quaternary amine strongly basic anion exchanger (capacity ~ 0.3 meq/g)• Not suited for very strong anions such as sulfonic acids, because these are difficult to elute

Recommended application:• organic acids• caffeine• saccharin

Cat. # Product Weight Size Qty/pkg730275 Chromabond®, SA 200mg 3ml 50730077 Chromabond®, SA 500mg 3ml 50730425 Chromabond®, SA 500 mg 6 ml 30731832 Chromafix®, SA 450 mg M 50

Sulfonamides in meat and kidneyB. Pacciarelli et al., Mitt Gebiete Lebensm. Hyg. 82 (1991) 45-55 Sample application: 1/10 of the extract volume, flow rateCompounds investigated: sulfaguanidine, sulfanilamide, sulfadiazine, about 2 mL/min; the column must not run drySulfathiazole, sulfapyridine, sulfamerazine, sulfamethizole, sulfadimidine, Column washing: 5 mL water, then 5 mL methanol; dry sulfamethozypryridazine, sulfachlorpyridazine, sulfadoxine, sulfadimethoxine for 10 min under vacuum. Now suck NH3 gas throughColumn type: the column until the acid is neutralized. To control theCHROMABOND® SA (= SCX), 3 mL, 500 mg neutralization process, press air through the column: a wet pH paper should indicate a neutral or basic pH value.Sample pretreatment: homogenize 10 g sample and 60 mL dichloromethane Elution: 3 mL methanol (1-2 mL/min)l carfully concentrate- acetone (1:1,v/v) for 30 s with a polytron. Centrifuge the homogenate for the eluate on a rotation evaporator (40 ºC/100 mbar)10 min at 2500 rpm. Filtrer the organix phase and wash the filter residue with a dissolve the residue in 0.5 mL of 5.5% acetonitrile inlittle dichloromethane - acetone. Add 5 mL glacial acetic acid to the filtered extract. buffer (1.641 g sodium acetate in 1 L water, adjusted to Column conditioning: apply 6 mL hexane and suck air until the column is dry pH 5 with glacial acetic acid) and centrifuge.(10 min). Then apply 6 mL dichloromethane - acetone - glacial acetic acid Further analysis: HPLC(10:10:1, v/v/v). Now the column must not run dry.

Vitamins: folic acid from food (e.g., wheat germs)Column type:CHROMABOND® SB (= SAX), 3 mL, 200 mg

Sample pretreatment: homogenize 10 g food sample in 100 mL0.01 mol/L phosphate buffer pH 7.4 and filterColumn conditioning: 2 column volumes n-hexane, then 2 column volumes methanol, finally 2 column volumes dist. waterSample application: force or aspirate 10 mL of the filtrate through the columnElution: 5 mL 10% sodium chloride in 0.1 mol/L sodium acetate buffer.

SPE


CHROMABOND® - QuEChERS Properties: • Base material silica, pore size 60 Å, particle size 45 μm, specific surface 500 m2/g, pH stability 2–8• Primary and Secondary Amine functions (PSA), 5 % C• Removes polar compounds (e.g., organic acids, pigments, sugars) from matrices like fruit or vegetables• Similar phases: Supelclean PSA, Bond Elut PSA

Method and Pre-Mixes Within a few years after its development by Anastassiades et al. the QuEChERS method has gained a leading position for determination of pesticide residues in food samples by GC-MS or LC-MS, allowing rapid and cheap clean-up of strongly matrix-contaminated samples. For optimizing the extraction of pH-dependent compounds, for minimizing decomposition of sensitive substances, and for broadening the matrix spectrum, different modifications of the QuEChERS method have been elaborated. In addition to the required adsorbent CHROMABOND® Diamino offers a number of individually weighed and premixed buffer and extraction mixtures, specially composed for different sample matrices.

The European standard EN 15662 recommends a citrate extraction mix (Mix I), while AOAC standard 2007.1 uses an acetate extraction mix (Mix II).

For clean-up, the Diamino phase (PSA) removes, e.g., sugars and organic acids. Magnesium sulfate removes water, C18 ec removes nonpolar interferences such as fats and the Carbon phase removes pigments, sterols, and nonpolar interferences.

Sample Property QuEChERS Extraction Mixes

QuEChERS clean-up mixes

Low fat content (e.g., Apples, Strawberries)

Citrate or acetate extractionMix I or Mix II

Diamino clean-upMix III

Moderate content of chlorophyll and carotinoids (e.g., Carrots, Lettuce)

Citrate or acetate extractionMix I or Mix II

Diamino/Carbon clean-upMix IV

Higher content of chlorophyll and carotinoids (e.g., Bell Peppers, Spinach)

Citrate extractionMix I

Diamino/Carbon clean-up (higher Carbon content)Mix V

Higher fat content(e.g., Avocado)

Citrate extractionMix I

Diamino/C18 ec clean-upMix VI

QuEChERS Mixes

Cat. # Description Composition Volume Qty/pkg730970 Mix I Citrate Extraction Mix 4 g MgSO4, 1 g NaCl, 0.5 g Na2H Citrate • 1.5 H2O,

1 g Na3 Citrate • 2 H2O15 ml 50

730971 Mix II Acetate Extraction Mix 6 g MgSO4, 1.5 g Na Citrate 15 ml 50

CHROMABOND® QuEChERS Exctraction Buffer Mixes

Cat. # Description Composition Volume Qty/pkg730972 Mix III Diamino Clean Up Mix 0.15 g CHROMABOND® Diamino with 0.9 g

MgSO4

15 ml 50

730973 Mix IV Diamino/Carbon Clean Up Mix 0.15 g CHROMABOND® Diamino with 0.9 g MgSO4 and 15 mg Carbon

15 ml 50

730974 Mix V Diamino/Carbon Clean Up Mix 0.15 g CHROMABOND® Diamino with 0.9 g MgSO4 and 45 mg Carbon

15 ml 50

730975 Mix VI Diamino/C18 ec Clean Up Mix 0.15 g CHROMABOND® Diamino with 0.9 g MgSO4 and 150 mg C18 ec

15 ml 50

CHROMABOND® QuEChERS Clean Up Mixes

Cat. # Description Weight Size Qty/pkg730561 Diamino Adsorbent 200 mg 3 ml 50730562 Diamino Adsorbent 500 mg 6 ml 30

CHROMABOND® Diamino Polypropylene SPE Cartridges

Cat. # Description Qty/pkg730653.20 Diamino Adsorbent 20 g730653 Diamino Adsorbent 100 g

CHROMABOND® Diamino Adsorbents

Recommended application:• Special SPE phase for quick and cheap determination of pesticides in strongly matrix-contaminated samples by GC-MS and LC-MS • QuEChERS method = Quick Easy Cheap Effective Rugged Safe

SPE


CHROMABOND® - XTRProperties: • Base material coarse-grained diatomaceous earth• Large pore size, high pore volume, constantly high batch-to-batch quality• pH working range 1–13• Fast, reproducible and economical• Simultaneous preparation of several samples• No problems with phase separation ∙ no formation of emulsions• High recovery rates• Saving of time and solvents• Organic solutions need not to be dried after separation

Recommended application:• Liquid-liquid extraction of highly viscous aqueous solutions such as physiological fluids (blood, plasma, and serum) in clinical chemistry, dyes in textiles, environmental and food analysis without use of a separation funnel• High water loadability without breakthrough of water during elution with organic solvents• Also suited for removing small amounts of water from solvents which are not miscible with water

Diatomaceous earth phase for liquid-liquid extractionDepending on the concentration of the analytes eluates can be analyzed immediately, or the organic solvent is evaporated. The pH value of the aqueous solution can be altered on the column, which enables elution of different compounds of a sample under optimized conditions. Under certain circumstances, acidic, neutral, and basic compounds can be fractionated in this way.

Solvents applicable for elution• diethyl ether • tert-butyl methyl ether • ethyl acetate• n-hexane • cyclohexane • toluene• dichloromethane • trichloromethane • trichloromethane – methanol • trichloromethane – methanol (85:15, v/v) • diethyl ether – ethanol (90:10, v/v) • diethyl ether – ethanol (80:20, v/v) • dichloromethane – 2-propanol (90:10, v/v)

Cat. # Product Weight Size Qty/pkg730507 Chromabond®, XTR 14.5g 70ml 30730487 Chromabond®, XTR 1g 6ml 30

Volume Amount of Adsorbent Max. volume capacity of aq. solution

Waiting period before elution

Elution volume

1 mL 250 mg 0.25 mL 5 min 3 mL3 mL 500 mg 0.5 mL 5 min 6 mL6 mL 1 g 1 mL 5–10 min 8 mL15 mL 3 g 3 mL 5–10 min 12 mL30 mL 4.5 g 5 mL 5–10 min 16 mL45 mL 8.3 g 10 mL 10–15 min 24 mL70 mL 14.5 g 20 mL 10–15 min 40 mL

150 mL 37.5 g 50 mL 10–15 min 90 mL

Sample Pretreatment:Weigh about 1 g cut - up textile sample (colored textiles about 0.1 g) in a 100 mL threaded vial. (Degrease leather samples before processing: cover sample with technical purity n-hexane and put the vial in an ultrasonic bath for 20 min. After decanting the n-hexane renise with little n hexane and dry sample by gentle heating and blowing with air or N2.) Add 250 μL internal standard (IS: 1.2mg/mL tetramethylbenzidine in methanol - ethyl acetate (1:1,v/v)), 17.0 mL citrate beffer (pH 6) (25.05 g citric acid and 12.64 g NaOH, fill up with deionized water to 2 L) and heat 30 min at 70 ºC. Then add 3 mL of a freshly prepared solution of 0.2 g/mL sodium dithionite in water and heat for exactly 30 min to 70 ºC while shaking occasionally.

Sample application:Cool the solution immediately (put vial in water - stoping of reductive cleavage). After 5 - 10 min pour it onto the CHROMABOND® XTR column (squeeze textile remains).

Determination of azo dyes and aromatic amines in colored textile materials with reference to § 64 LFGB (Formerly § 35 LMBG) Elution:Allow solution to be soaked up by the adsorbent for 15 min. Then elute four times with 20 mL each of diethyl ether or diethyl ether - ethenol (90:10, v/v) (depending on recovery rates), using the first 40 mL to rinse the sample remains. Evaporate eluates to 3 mL with a rotation evaporator and transfer the solution into a 10 mL measuring flask using a pasteur pipette and rinsing with methanol. Fill up to the marking with methanol, shake, and pipette about 1 mL into a vial.

Further analysis:Fast GC on OPTIMA® §-3, 10 m, 0.1 mm ID, 0.1 μm filmHPLC column, Nucleosil® 100-5 C18 HD (App. # 110500)

SPE


CHROMABOND® - Vacuum Manifold for 12 columnsFor simultaneous preparation of up to 12, 16 or 24 sample. Replacement parts and accessories for special applications

1. Rectangular glass cabinet; 2 sizes available: • small for up to 12 CHROMABOND ® columns or CHROMAFIX® cartridges; • large for up to 16 CHROMABOND ® LV columns or up to 24 CHROMABOND ® columns or CHROMAFIX ® cartridges (depending on lid)

2. Polypropylene lid3. Vacuum gauge for pressure reading4. Control valve for adjustment of vacuum 5. Replaceable valves for vacuum control of individual SPE columns6. Variable rack with exchangeable partitions, which accept a wide variety of vessels like test tubes, measuring flasks, scintillation vials, autosampler vials, plastic vials etc.7. CHROMABOND ® LV columns with 15 mL sample reservoir for medium size samples8. Polypropylene sample reservoir (30 or 70 mL available)9. Adapter for sample reservoirs10. CHROMABOND ® tubing adapters

Protection From Cross ContaminationFor special applications, which require maximum protection from cross contamination we supply chrome-plated brass valves and stainless steel or PTFE connectors, the application of which is shown below. These special connectors are fitted through the lid; thus the sample only has contact with the inert connector and can flow directly into the receptacle.

Drying AttachmentIf the eluate has to be evaporated, this can be performed with the so-called drying attachment (11, see below). This special lid has a gas connector on one side (12), from which the gas is fed simultaneously to the 12 or 24 stations (13). Thus 12 or 24 eluates can be evaporated simultaneously by just changing the lid and applying a stream of inert gas, e.g., nitrogen.

SPE Column with fritsand solid phase

Luer fitting, female

Lid of vacuum manifoldwith bore

Luer fitting, male

Connector

Stainless steel needle

SPE


Cat. # Description Qty/pkg730150 Vacuum Manifold Complete for up to 12 columns or cartridges (including PP tank) 1730360 Vacuum Manifold Complete for up to 16 LV columns 1730151 Vacuum Manifold Complete for up to 24 columns or cartridges 1

CHROMABOND® Vacuum Manifolds - Complete

Cat. # Description Qty/pkg730173 Glass Cabinets without Accessories for 12 Columns 1730174 Glass Cabinets without Accessories for 16 LV or 24 Columns 1

CHROMABOND ® Vacuum Manifolds - Glass Cabinets without Accessories

Cat. # Description Qty/pkg730175 Lids with Gaskets for 12 columns (including Luer fittings and values) 1730365 Lids with Gaskets for 16 LV columns (including Luer fittings and values) 1730176 Lids with Gaskets for 24 columns (including Luer fittings and values) 1730177 Gaskets for Lids, for 12 columns 2730178 Gaskets for Lids, for 24 columns 2

CHROMABOND® Vacuum Manifolds - Lids with Gaskets

Cat. # Description Qty/pkg730194 Luer Stoppers for Vacuum Manifold, Blue 12730183.12 Luer Fittings for Lid, Female 12730184.12 Luer Fittings for Lid, Male 12730185 Valves, Plastic 12730152 Stainless Steel Needles 12730154 Polyproylene Needles 12730233 PP tanks for Vacuum Manifold for 12 Columns (Not Available for 16 or 24 Position Manifolds) 2730179 Vacuum Gauge, Complete with Accessories 1

CHROMABOND® General Accessories for Vacuum Manifolds

Cat. # Description Qty/pkg730187 Dry Attachments for 12 Columns 1730188 Dry Attachments for 24 Columns 1730157 Collecting Rack for 12 Columns 1730366 Collecting Rack for 16 LV Columns 1730153 Collecting Rack for 24 Columns 1

CHROMABOND® Drying Attachments and Collection Racks - For Evaporation of Eluates

Cat. # Description Qty/pkg730189.1 Valve Brass Tarnish 1730189.12 Valves, as Above 12730106 Stainless Steel Connectors 12730564 PTFE Connectors 12730563 PTFE Connectors with Valve 12

CHROMABOND® Product for Protection from Cross Contamination

Cat. # Description Qty/pkg730387 Tubing Adaptors for Application of Large Sample Volumes For 1, 3, and 6 mL Glass Columns 4730243 Tubing Adaptors for Application of Large Sample Volumes For 1, 3, and 6 mL Polypropylene Columns 4730386 Tubing Adaptors for Application of Large Sample Volumes For 15, 45, and 70 mL Polypropylene Columns 4

CHROMABOND® Tubing Adaptors for Application of Large Sample Volumes

SPE


SPE Cartridges - JordiJordi brings you the highest quality 100% divinylbenzene (DVB) cartridges available. These resins are designed with a 10 μm average particle size compared to the typical 40 μm of leading brands. This smaller particle size provides extremely high retention efficiency and higher capacity. Jordi cartridges offer a significantly larger useful surface area while maintaining excellent flow characteristics.

Jordi offers the broadest range of polymer-based surface chemistries on the market, addressing aqueous, organic and ion exchange sample preparation. Jordi resins are made from 100% divinylbenzene, making them more retentive than standard PS-DVB packings. This makes Jordi Hydroclean SPE one of the most retentive phases available, providing increased analyte recoveries.

Jordi Quality Assurance

All Jordi SPE products are manufactured with the highest quality DVB and checked by stringent standards. Quality control for all of Jordi’s materials include:

• Particle Size - Electrozone sensing analysis of particle size distribution• Particle Shape - Light microscopy analysis of particle shape and size• Surface area and Porosity - Nitrogen adsorption• Chromatographic performance - Column chromatography• Purity - Mass spectroscopy (MS)• Phases are availabe in powder form in bulk

Advantages of Polymeric SPE• Outstanding recovery due to the highly retentive nature of polymeric phases• Absence of leachables from the polymeric media since there is no bonded phase• Lack of residual silanol groups• Incredible ruggedness of polymeric supports allows wide pH and solvent compatibility• Improved batch to batch reproducibility• Higher capacity for improved detection limits and reduced resin volumes

Hydroclean RP• Localized hydrophobic/hydrophilic balance for optimal analyte/resin contact and retention• Patent pending polyamide stationary phase allows for NP and RP separations in 100% water or 100% organic solvent• Exhibits improved wettability even after drying

Hydroclean SCX• Sulfonated stationary phase allows for cation exchange separations• 100% DVB supports promote increased durability, excellent flow properties and stability throughout the entire pH scale• SCX exhibits excellent wettability even after drying due to hydrophilic/hydrophobic balance

Hydroclean SAX• Quaternary amine phase allows for anion exchange separations• Features 100% DVB supports, for enhanced retention characteristics• SAX exhibits excellent wettability even after drying due to hydrophilic/hydrophobic balance

Hydroclean RP DVB• 100% Divinylbenzene phase• Features enhanced retention over styrene/DVB phases• DVB – the phase of choice for highest retention

SPE


Cat. # Description Pore Size Part. Size Size Bed Weight Qty/pkg500SPE-001-00030 Hydroclean SCX 80 Å 25-35 μm 1 ml 30 mg 100500SPE-003-00060 Hydroclean SCX 80 Å 25-35 μm 3 ml 60 mg 100500SPE-006-00200 Hydroclean SCX 80 Å 25-35 μm 6 ml 200 mg 30500SPE-961-00030 Hydroclean SCX - 96 Well 80 Å 25-35 μm 1 ml 30 mg 1510SPE-001-00030 Hydroclean RP DVB 80 Å 25-35 μm 1 ml 30 mg 100510SPE-003-00060 Hydroclean RP DVB 80 Å 25-35 μm 3 ml 60 mg 100510SPE-006-00200 Hydroclean RP DVB 80 Å 25-35 μm 6 ml 200 mg 30510SPE-961-00030 Hydroclean RP DVB - 96 Well 80 Å 25-35 μm 1 ml 30 mg 1530SPE-001-00030 Hydroclean RP 80 Å 25-35 μm 1 ml 30 mg 100530SPE-003-00060 Hydroclean RP 80 Å 25-35 μm 3 ml 60 mg 100530SPE-006-00200 Hydroclean RP 80 Å 25-35 μm 6 ml 200 mg 30530SPE-961-00030 Hydroclean RP - 96 Well 80 Å 25-35 μm 1 ml 30 mg 1580SPE-001-00030 Hydroclean SAX 80 Å 25-35 μm 1 ml 30 mg 100580SPE-003-00060 Hydroclean SAX 80 Å 25-35 μm 3 ml 60 mg 100580SPE-006-00200 Hydroclean SAX 80 Å 25-35 μm 6 ml 200 mg 30580SPE-961-00030 Hydroclean SAX - 96 Well 80 Å 25-35 μm 1 ml 30 mg 1

Jordi SPE Cartridges


Melamine with Milk Plasticizers and UV Screens

ProcedureProduct: HRP-SCX – 200 mg 6 mlCat. #: 500SPE-006-00200Cartridge Prep: Solvent 1 – Methanol (3 ml) Solvent 3 – Hydrochloric Acid (3 ml, 0.1 M)Sample Loading: 4 ml of total solutionWash: Wash 1 – Hydrochloric Acid (3 ml, 0.1M) Wash 2 – (75/25) Methanol / Hydrochloric Acid (0.1M) (3 ml) Dry Step – 10 mm Mercury (~ 5 min.)Elution: (95:5) Methanol / NH4OH (3 ml)Matrix: Milk diluted 3:1 with 0.1M HCl Melamine at 0.25 mg/ml

Detection MethodPacking: Jordi Gel DVB 500 ÅColumn: 4.6 mm x 150 mmCat. #: 16502Mobile Phase: 10/20/30/40 MeOH/CAN/THF/H2OFlow Rate: 1 ml/minDetection: UV238nm

Analyte: MelamineConc.: 1 mg/mlRecovery: 93.1 %

ProcedureProduct: HRP – 200 mg 6 ml Cat. #: 530SPE-006-00200Sample Loading: The solution was completely loaded and was allowed to pull to drynessWash: Wash Solvent 1 – Methanol (2 ml) Wash Solvent 2 – (75/25) Water Methanol (3 ml) Elution: 2 ml Methanol

Analytes ppm(1/25 dilution) % RecoveryTriethylcitrate 35 98Diethylphthalate 30 106Dioctylphthalate 26 1006-Undecanone 29 102Hexadecanol 34 109Padimate O 33 79

SPE

Solid Phase Extraction (SPE) - Sorbent Technologies · Solid Phase Extraction (SPE) is a powerful...

Documents

Transcript of Solid Phase Extraction (SPE) - Sorbent Technologies · Solid Phase Extraction (SPE) is a powerful...