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Automated Sample Preparation of 25-OH-Vitamin D3/D2 from Serum with the AC Extraction Plate and the Tecan Freedom EVO® for the quantification by LC-MS/MS

5 Mar 2014

This application note describes an automated method for the determination of Vitamin D in serum using LC-MS/MS analysis. Details of the workflow are described.

AC Extraction Plate™

Tecan

The Tecan AC Extraction Plate* with TICE™ (Tecan Immobilized Coating Extraction) technology revolutionizes your sample preparation routine. The AC Extraction Plate is a 96-well, deep-well microplate, with each well having a proprietary TICE™ coating. TICE is a material with distinct absorptive properties, and this coating efficiently extracts specific small molecule analytes from aqueous solutions, releasing them when in contact with a suitable organic solvent. A simple pipette and shake sequence, with no filtration, centrifugation or solvent evaporation, is all that is required. The AC Extraction Plate is easily integrated into automated processes, making it a perfect match with Tecan’s Freedom EVO® liquid handling platform. AC Extraction Plate™ Features: Cost-effective – Simple pipette and shake workflow, without labor intensive centrifugation, filtration and solvent evaporation steps. Convenient and fast – Just unpack and start, no conditioning or other preparations needed. Highly reproducible – Pipette and shake routines are easily standardized and automated on a liquid handling platform, eliminating random errors and increasing consistency of results. * For research use only – not for clinical diagnostic applications

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Te-Shake™

Tecan

Tecan’s Te-Shake orbital mixer offers software-controlled shaking and heating options for microplates, deep-well plates and microfuge tubes. Te-Shake is fully compatible with Tecan’s pipetting platforms, providing automated transport of plates to and from the shaker platform. With multiple configurations available, Te-Shake is sure to fit your laboratory’s specific application needs. Te-Shake is easily integrated into Freedom EVO® or Genesis-based workstations. No external power supply is necessary. Straightforward operation of the device is provided by Tecan’s Pipetting Software Package. On-deck mounting optionally allows pipetting during or between shaking periods. Due to flexible shaking speed and rotation mode, optimal mixing results will always be achieved even for delicate resuspension tasks. Te-Shake highlights: Te-Shake can accommodate virtually any type of microplate or deep-well block The module is available in various configurations that address, for example, heated mixing and/or multiple parallel microplate shaking With its dynamic shaking range, configurable orbit/amplitude and variable rotating direction, Te-Shake allows easy optimization of extended mixing Automated exchange of plates on Te-Shake allows multiple batch processes without user intervention Process scale-up can be realized by parallel use of multiple Te-Shake options on one single workstation Te-Shake applications: Cell lysis processes Bead-based extraction methods Cell metabolism assays Solubilization and homogenization tasks

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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. Robotic WorkstationsRobotic workstations are automated platforms enabling the user to perform high-throughput, multi-application processes, from sample preparation tasks such as PCR set-up, DNA purification and NGS library preparation, to integrated sample prep and analysis workstations. Automation makes processes consistent, fast, precise and fully walk-away. Find the best robotic workstations in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Automation Supplies / AccessoriesAutomated accessories and consumables make lab processes fast, efficient and reliable. Useful automation accessories include microplate sealers, label printers and reagent reservoirs.Automated Liquid HandlingAutomated liquid handling equipment is used to mix, dilute and dispense allotted volumes of liquid into microplates and other vessels automatically. The robotic, liquid handling arms can dispense from single channel to 3456 multichannel as well as operate nanoliter dispensing, enabling high throughput of samples. Find the best automated liquid handlers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Vitamin D