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Therefore, typical sample matrices that can be employed in normal-phase SPE include hydrocarbon or fatty oils diluted in an organic solvent, hexane, isooctane, chlorinated solvents, THF, diethyl ether, and ethyl acetate. Most organic analytes exhibit some polar functionalities that can be exploited for normal-phase separation. Because many molecules exhibit polar functionality, each interaction can provide different levels of selectivity offering highly selective separations of compounds very similar in structure. Normal-Phase SPE Retention Mechanism: Polar Interactions - Hydrogen bonding, π-π, dipole-dipole, and induced dipole-dipole Sample Matrix: Non-polar samples - Organic extracts of solids - Very non-polar solvents - Fatty oils, hydrocarbons Analyte Characteristics: Analytes exhibiting polar functionalities - Hydroxyl groups, carbonyls, amines, double bonds - Hetero atoms (O, N, S, P) - Functional groups with resonance properties Elution Scheme: Polar interactions disrupted with a more polar solvent or solution - Acetonitrile, methanol, isopropanol - Combinations of buffer/solvent or solvent/solvent mixtures Common Applications: - Cleanup of organic extracts of soils and sludge - Fractionation of petroleum hydrocarbons - PCBs in transformer oil - Isolation of compounds in cosmetics ## Basic Steps 1. Sample Pre-treatment Liquid samples should be initially extracted or diluted with a non-polar solvent such as hexane or a chlorinated solvent. Soil, sediment, and other solid samples are initially extracted (Soxhlet or sonication) with a non-polar solvent, and concentrated prior to SPE cleanup. Aqueous residues in the sample can reduce normal-phase retention. It may be necessary to further dry the organic extract with sodium sulfate or magnesium sulfate prior to SPE. 2. Conditioning/Equilibration Condition and equilibriate with 2-3 tube volumes of a non-polar solvent similar or identical to sample matrix resulting from sample pre-treatment. 3. Sample Load Apply sample (from step 1) at a consistent and reduced flow rate of ~1-2 drops/second to ensure optimal retention. The compounds should be a non-polar solvent (e.g., hexane) for optimal retention. Note that methanol and acetonitrile are often used as elution solvents in normal-phase SPE, and will often not promote compound retention during sample load. 4. Wash Sample interferences are often co-retained with compounds of interest during sample load. A wash step is necessary to elute interferences without prematurely eluting compounds of interest. In normal-phase SPE, 1-2 tube volumes of the solvent used in sample pre-treatment and conditioning can be used during wash. 5. Elution Disrupt polar interactions with a solvent or solvent/buffer mixture more polar than both the sample and wash solutions. Typical elution solvents include water miscible organic solvents such as acetone, acetonitrile, methanol, and isopropanol. Eluting with increasingly polar solvents (or solvent mixtures) in succession can also fractionate multiple compound classes. See “Common Normal-Phase Solvents” table for assistance. 6. Eluate Post-treatment Normal-phase SPE is often followed by GC analysis, and therefore requires a volatile sample matrix prior to injection. Use sodium sulfate or magnesium sample to remove residual moisture. Further SPE eluate concentration may also be necessary prior to analysis. ### Common Normal-Phase Solvents lventElutropic (e°) or elution strength on silicaPromotes Normal-Phase Retention / Elution Hexane0.00__Retention__ Isooctane0.00 Carbon tetrachloride0.14 Toluene0.22 Benzene0.27 Tert-butyl methyl ether0.29 Chloroform0.31 Methylene chloride (dichloromethane)0.32 Diethyl ether0.29 Ethyl acetate0.43 Tetrahydrofuran0.35 Acetone0.45 Acetonitrile0.50 40% methanol in acetonitrile0.67 20% methanol in diethyl ether0.65 20% methanol in methylene chloride0.63 Isopropanol0.63 Methanol0.73 Water>0.73__Elution__ Acetic acid>0.73 __Related Technical Articles__ - [Determination of Formaldehyde and Acetaldehyde in Air Using DNPH Cartridges and Automated On-Line Desorption Followed by HPLC](https://www.sigmaaldrich.com/KR/en/technical-documents/protocol/environmental-testing-and-industrial-hygiene/air-testing/aldehydes-air-dnph) - [Effect of Container Materials on Milk Off-Flavors](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/gas-chromatography/container-materials) - [Pesticide Residue Analysis in Ginger Powder with QuEChERS](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/quechers/analysis-of-pesticide-residues-in-ginger-powder) - [HybridSPE-Phospholipid Technology](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/small-molecule-hplc/hybridspe-precipitation-technology) - [Impact of Mobile Phase Additives on LC-MS Sensitivity, Demonstrated using Spice Cannabinoids](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/small-molecule-hplc/lcms-spice-cannabinoids) - [Analysis of Drugs of Abuse in Urine](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/solid-phase-extraction/analysis-of-drugs-of-abuse-in-urine-after-cleanup-with-hlb) - [LC-MS Analysis of Warfarin™ in Plasma Samples](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/solid-phase-extraction/chiral-and-achiral-lc-ms-analysis-of-warfarin-in-plasma-samples) - [QuEChERS Cleanup and Analysis of Veterinary Drugs in Milk by LC/MS/MS](https://www.sigmaaldrich.com/KR/en/technical-documents/technical-article/analytical-chemistry/solid-phase-extraction/dispersive-spe-quechers-veterinary-drugs-milk) - [View More](https://www.sigmaaldrich.com/KR/en/search/facet-search?focus=sitecontent&term=facet-search) 상부 __계속하려면 로그인하세요.__ 계속 읽으시려면 로그인하거나 계정을 생성하세요. 로그인__계정이 없으십니까?__등록 지원[고객 지원](https://www.sigmaaldrich.com/KR/ko/support/customer-support)[문의하기](https://www.sigmaaldrich.com/KR/ko/collections/offices)[자주 묻는 질문](https://www.sigmaaldrich.com/KR/ko/life-science/ecommerce/ecommerce-faq)[안전 데이터 시트(SDS)](https://www.sigmaaldrich.com/KR/ko/documents-search?tab=sds)[인증서(COA/COO)](https://www.sigmaaldrich.com/KR/ko/documents-search?tab=coa)[품질 및 규제](https://www.sigmaaldrich.com/KR/ko/life-science/quality-and-regulatory-management)[계산기 및 앱](https://www.sigmaaldrich.com/KR/ko/support/calculators-and-apps)[웹 세미나](https://www.sigmaaldrich.com/KR/ko/collections/webinars) 주문[주문 조회](https://www.sigmaaldrich.com/KR/ko/order-lookup)[빠른 주문](https://www.sigmaaldrich.com/KR/ko/quick-order)[맞춤형 제품](https://www.sigmaaldrich.com/KR/ko/services/custom-products)[전자상거래 솔루션](https://www.sigmaaldrich.com/KR/ko/life-science/ecommerce/ecommerce-solutions) 회사[회사 소개](https://www.sigmaaldrich.com/KR/ko/life-science/about-us)[책임](https://www.sigmaaldrich.com/KR/ko/life-science/ssbi)[이벤트](https://www.sigmaaldrich.com/KR/ko/collections/events)[보도 자료](https://www.sigmaaldrich.com/KR/ko/collections/press)[프로그램](https://www.sigmaaldrich.com/KR/ko/life-science/partnership-programs)[채용 정보](https://careers.merckgroup.com/global/en)[사무실](https://www.sigmaaldrich.com/KR/ko/collections/offices) 소셜 미디어 [![LinkedIn icon](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/images/icons/linkedin.svg)](https://www.linkedin.com/company/merck-life-science) [![X icon](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/images/icons/x-icon.svg)](https://x.com/Merck_lifesci) [![Facebook Icon](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/images/icons/facebook.svg)](https://www.facebook.com/Merck.lifescience) [![Instagram Icon](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/images/icons/instagram.svg)](https://www.instagram.com/mercklifescience/) [![YouTube Icon](https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/images/icons/youtube.svg)](https://www.youtube.com/@MerckLifeScience/featured) Merck __연구. 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