Solid-Phase Extraction with Oasisfi HLB Sorbent

1 Solid-Phase Extraction withOasis HLB Sorbent :Simple Procedures for SuperiorSample 1998 Waters Presents:Outline Introduction Oasis HLB Sorbent Physical properties Advantages compared with traditionalsorbents A Simple Strategy for Most Samples Agrochemicals and metabolites Pharmaceuticals Environmental Advanced Strategies for DifficultSamples 1998 Waters phase Extraction (SPE) methods are widely used for organic analysis of aqueous samples in the agrochemical, pharmaceutical, and environmental industries. Traditional silica based sorbents - effective for non-polar analytes Polymeric sorbents - higher capacity than silica based sorbentsOasis HLB - more effective for polar analytes and metabolites 1998 Waters of TraditionalSample Preparation Methods Liquid-Liquid Extraction Cumbersome Time consuming Not easy to automate Not good for polar drugs and metabolites Solid-Phase Extraction Overcomes many problems of othermethods, but.

2 1998 Waters of Silica-Based SPE SorbentsDifficult to optimize for polar compounds Inconsistant recovery Lower capacity Tedious Extractions require close attention socartridges do not run dry 1998 Waters Ideal Sample PreparationMethod: Removes interferences High and reproducible recoveries for acidic,basic and neutral analytes with a broad range ofpolarities Easy to automate for high sample throughput Easy to use and rugged Fast and cost efficient 1998 Waters HLB Extraction products contain a resinmade from a co-polymer of divinybenzene andvinyl pyrrolidinone.

3 The pyrrolidinonefunctionality acts as an imbedded hydrophilicgroup. The pyrrolidinone functionality alsoprovides enhanced retention for some this seminar, applications are presented forthe agrochemical, pharmaceutical, andenvironmental industries 1998 Waters Corp. Provides wetting properties Reduces contact angle withwater Provides enhanced retention forpolars Provides reversed-phaseproperty for analyte retentionHydrophilicmonomerLipophilicmon omerNOOasis HLB Sorbent :A Hydrophilic-Lipophilic BalancedCopolymer 1998 Waters Properties of Oasis HLB Sorbent Specific Surface Area:800 m2/g Average Pore Diameter:80 Specific Pore mL/g Average Particle Diameter.

4 30 m **60 m particles in products with more than 200 mg Sorbent 1998 Waters of Drying on Recovery withSilica-based CartridgeBouvier, CapparellaProcainamideAcetaminophenRanit idinePropranolol010203040506070809010002 468 Drying Time @ 10" Hg [min]Percent RecoveryDoxepin 1998 Waters :Water vs Methanol on C18-SilicaWater on C18d = 100 = dynes/cm = **B. Janczuk, T. Bialopiotrowicz and W. Wojcik, Colloids and Surfaces 36 (1989) 391-403 Methanol on C18d = 100 = 22 dynes/cm = * Pc ~ -1000 psi Pc ~ 1500 psi 1998 Waters : C18 vs.

5 Oasis HLB HLBC18H = hydrophilic unitL = lipophilic unitLLLLLLLLLLLLLLLLLLHHHHHH 1998 Waters Effect on Recovery:C18 vs. Oasis HLB CartridgesProcainamide Acetaminophen Ranitidine Propranolol DoxepinBouvier,Capparella020406080100051 0 Drying Time [min]0204060801000510 Drying Time [min]% Recovery% RecoveryC18 Oasis HLB 1998 Waters of Oasis HLB Sorbent High Capacity - sample size of 75 - 250 mL on 60 mgcartridge, 1 L on 200 mg cartridge Low elution volumes (~1 mL for 60 mg cartridge) Higher recovery for polar metabolites such ashydroxyatrazine compared with silica-based sorbents Proprietary cleaning process - no significant cartridgeinterferences for GC/NPD:ECD or HPLC/PDA (200-360nm) Detection limits of 100 ng/L or better No stopcocks needed for routine analysis 1998 Waters Corp.

6 Old Method, C18 Silica 1000 mg cartridge 500 mL sample, 50 minutes final volume 4 mL recovery > 85% LOQ ~ 50 ng/L if bed runs dry duringconditioning or loading, startover New Method, Oasis HLB 60 mg cartridge 100 mL sample, 15 minutes final volume 1 mL recovery > 90% LOQ ~ 50 ng/L do not worry if bed runs dryduring conditioning or loadingExample: Sulfonylurea HerbicidesSHNOOCHNONNNOCH3CH3 SCO2CH3 Thifensulfuron methylClSHNOOCHNONNNOCH3CH3 ChlorsulfuronCO2CH3 SHNOOCHNONNNOCH3CH3 Metsulfuron methyl 1998 Waters Types Typical Analytes Sample SizePharmaceuticaldrugs, metabolites1-10mLplasma, urineForensicdrugs of abuse1-25mLurine, tissue extractsEnvironmental organic pollutants 25mL-1 Lwater, soilAgrochemicalpesticides, metabolites 1mL-1 Lwater, soil, tissue,plasma, urineFoodnutrients.

7 Pesticides1-100mLbeveragesMajor Industries Using SPE 1998 Waters of Cartridge Based onSample SizeSample Size Cartridge Size(clean samples)(dirty samples)1-25 mL1 cc, 30 mg3 cc, 60 mg25-200mL3 cc, 60 mg6 cc, 200 mg200-500mL6 cc, 200 mg6 cc, 500 mg (LP)500-1000mL6 cc, 500 mg (LP) 12 cc, 1g (LP) 1998 Waters SPE Method for HPLC Analysis(conditions for 3 cc, 60 mg Oasis HLB cartridge)Condition/Equilibrate1 mL methanol/1 mL waterLoadup to 200 mL sampleWash1 mL 5% methanol in waterElute1 mL methanolEvaporate and ReconstitutePrepare SampleWhen analyzing forbasic compounds, do notuse high organic contentwash.

8 There are nosilanols on Oasis the general methodfirst. Use 2-D method ifinterference reductionis required. 1998 Waters SPE Method for GC Analysis(conditions for 6 cc, 200 mg Oasis HLB cartridge)3 mL solvent*/3 mL methanol/3 mL waterDry (Na2SO4), Adjust to Final VolumeCondition/EquilibrateLoadup to 500 mL sampleWash3 mL 5% methanol in waterElute6 mL solvent*Prepare Sample*typical solvents: ethyl acetate, MTBE, methylene chloride 1998 Waters Agrochemical, Environmental triazine herbicides and metabolites PAHs acidic herbicides and phenols Pharmaceutical tetracyclines basic drugs (1-D and 2-D) Food tetracyclines pesticides 1998 Waters , we will look at some applications using the general methods.

9 The general methods are simple and one dimensional (1-D, no pH adjustment in wash/elute steps).Agrochemical: Organonitrogen herbicides and metabolitesPharmaceutical and Food:TetracyclinesEnvironmental:Organoph osphorous Pesticides 1998 Waters cartridgespiked well water(200 ng/L)nonspiked sample1: Desisopropylatrazine2: Hydroxyatrazine3: Desethylatrazine4: Simazine5: Cyanazine6: AtrazineTriazine Herbicides and Metabolites - HPLCHPLC MethodColumn:SymmetryShield RP8 Mobile Phase:A: 15% Acetonitrile in pH buffer (5 mM)B: Acetonitrile Gradient.

10 100% A for 2 min, then lineargradient to 70% B in 25 minFlow mL/minDetection:UV @ 214 nm ( AUFS)Sample:75 mL Injection:75 LatrazineNNNClNHCH(CH3)2C2H5 NHSPE: 3 cc, 60 mg Oasis HLB 1998 Waters are given as % recovery with % RSD in parenthesisTap WaterWell WaterSpike level 500 ng/LSpike level 200 ng/LCompound5 replicates7 replicatesDesisopropylatrazine ( ) ( )Hydroxyatrazine132 ( )109 (11)Desethylatrazine106 ( )104 ( ) * ( ) * ( )Atrazine101 ( )101 ( )* Simazine and cyanazine were not included in the tap water spikeexperimentTriazine Herbicides Results 1998 Waters Herbicides and MetabolitesHPLC1: 2-[(2-ethyl-6-methylphenyl)amino]- 1-propanol2.