ResourceSeparations

In-well hydrolysis using UCT Refine Ultra-Filtration Technology

15 Mar 2023

In this application note, UCT outlines a suggested procedure for in-well hydrolysis of urine samples using the Refine Ultra-Filtration Technology. Various biological sample types commonly contain proteins that complicate downstream analysis if not effectively removed. While sample preparation techniques, such as solid phase extraction (SPE), can successfully remove these unwanted matrix components, complex and lengthy extraction procedures are not ideal for high-throughput laboratories. With the advancement of LC-MS/MS technology, many laboratories have implemented dilute and shoot procedures in order to reduce sample preparation time, reduce consumable spending and reduce overall turnaround time. The caveat to this quick protocol is the toll that it ultimately takes on the HPLC column and mass spec source.

The Refine Ultra-Filtration Technology allows for sample precipitation and filtration to occur simultaneously in the individual column/well. A novel, hydrophobically treated, submicron frit combination facilitates the removal of sample proteins without a complicated extraction. Using the Refine Ultra-Filtration Technology reduces column backpressure while also reducing analyte suppression when compared to a standard dilute and shoot protocol.

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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.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. UltrafiltrationHPLC Method DevelopmentIt is important to develop and validate HPLC methods in order to identify, quantify or purify compounds of interest in the most efficient way. Effective method development ensures optimization, quality control and cost effectiveness.HPLCHigh-performance liquid chromatography (HPLC) is a technique for separating, identifying, and quantifying components in mixtures. It is widely used in chemical, pharmaceutical, and environmental analysis to ensure purity and identify unknown compounds. Explore the best HPLC systems in our peer-reviewed product directory; compare products, read reviews, and get pricing directly from manufacturers.
In-well hydrolysis using UCT Refine Ultra-Filtration Technology