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Purifying and Concentrating Imiquimod and Metabolites from Large Volumes of Urine

3M Bioanalytical
4 Apr 2010
Sarah Sarah
Marketing / Sales

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This application note demonstrates the use of large diameter (47 mm) Empore Extraction Disks to purify and concentrate a novel immunomodulator drug, Imiquimod (AldaraTM), and its metabolites from urine. The eluates were analyzed by different chromatographic techniques (HPLC and TLC) to determine the nature and number of metabolites present.

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3M™ Empore™ C18 47 mm Disk

3M Bioanalytical

3M Bioanalytical's Empore ci8 47mm Disk - Commonly used for the solid phase extraction of analytes from complex sample matrices.  A variety of functional groups, such as octadecyl (C18) and octyl (C8) can be bonded to the silica surface to provide non polar interactions.  Each of these sorbents exhibits unique properties of retention and selectivity for a particular analyte.  The choice of which sorbent is best for a particular method will be influenced by the percent recovery of analyte from the sample matrix and the cleanliness of the resulting chromatography.

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3M™ Empore™ SDB-RPS 47 mm Disk

3M Bioanalytical

SDB-RPS is a poly(styrenedivinylbenzene) copolymer that has been modified with sulfonic acid groups to make it hydrophilic.  SDB-RPS is not bonded to silica but is a 100% copolymeric particle that is spherical, porous and cross-linked.  The sulfonation imparts unique selectivity for organic analytes that are polar in nature, such as drugs, drug metabolites, water-soluble pesticides and pesticide metabolites.  SDB-RPS is also useful for comprehensive drug screening, in which a wide range of analytes are simultaneously extracted.  SDB-RPS can be used as a reversed phase sorbent and substituted into existing methods.  Organic analytes with polar character have been shown to extract better on SDB-RPS than on traditional C18 bonded silica.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Solid-Phase ExtractionSolid-phase extraction (SPE) is used for clean-up, extraction and concentration of semi-volatile or non-volatile analytes from complex mixtures including blood, urine and food samples. Multiple formats are available for conducting SPE, including prepacked SPE cartridges, disks and microplates, as well as SPE sorbent powders for manual packing. SPE systems can be used to automate the process and extract multiple samples at once. Solid-phase microextraction (SPME) and supported liquid extraction (SLE) products are also available in the same formats. Find the best SPE, SLE and SPME equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Thin Layer ChromatographyThin-layer chromatography (TLC), also called planar chromatography, is an inexpensive and simple liquid chromatographic technique used to separate and identify small amounts of compound in a mixture, monitor progress of a reaction or determine purity of substance. TLC plates come in multiple sizes with common adsorbent layers being silica, aluminum oxide or cellulose and high-performance (HP-TLC) plates are also available for increased sensitivity. Other equipment needed includes TLC developing chambers, applicators, TLC sprayers and visualizing devices, such as a viewing cabinet and UV lamp. Find the best TLC products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.GC-MS GC-MS (gas chromatography-mass spectrometry) instruments and equipment are used to separate, quantify and identify mixtures of small and volatile compounds, such as polycyclic aromatics, fatty acids and alcohols. Often used in drug detection, forensic investigation and environmental analysis for pesticides and contaminants, GC-MS is a powerful addition to your lab’s analytical capabilities. GC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzers can give greater sensitivity and resolution to your analysis. Find the best GC-MS instruments and equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.

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Purifying and Concentrating Imiquimod and Metabolites from Large Volumes of Urine