Total Drug Analysis for Pharmacokinetic Studies
MultiScreen® Deep Well and MultiScreen Solvinert Filter Plates
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Overview
Specifications
Ordering Information
Documentation
References
| Reference overview | Application |
|---|---|
| LC–MS-MS Total Drug Analysis of Biological Samples Using a High-Throughput Protein Precipitation Method Bogdan Sleczka, Jian Wang, and Timothy Olah, Bristol-Myers Squibb Bioanalytical and Discovery Analytical Sciences, Princeton, New Jersey LC-GC, Volume 24 Number 7 July 2006 2005 | |
| Assessment of filter plates for multi-well in-gel digestion of proteins separated by polyacrylamide gel elelctrophoresis to identify them with LC-ESI/MSMS Kazuhisa Kameyama, Tomohiro Nanri, Yuko Yamanaka, Mikikio Arima, Hiroshi Kawasaki Journal Electrophoresis. 2005; 49, 71-75 2004 | Mass Spectrometry |
FAQ
| Question | Answer |
|---|---|
| What is the maximum volume that should be used with the MultiScreen Deep Well filter plate? | The maximum recommended volume to be used with the MultiScreen Deep Well filter plate is 1.8 mL (1.5 mL with shaking). |
| Should the MultiScreen Solvinert Deep Well plate be used with vacuum or centrifuge? | The MultiScreen Solvinert Deep Well plate with both vacuum and centrifuge. However, vacuum should not be used with volitile solvents (e.g., TFA). |
| At what g force should the MultiScreen Solvinert Deep Well plates be spun? | The MultiScreen Solvinert Deep Well plates should be spun at 500-3,000 x g. |
| What is the active membrane surface area of the MultiScreen Solvinert Deep Well plate? | The active membrane surface area of the MultiScreen Solvinert Deep Well plate is 0.28cm2. |
| What is the extractables level for the MultiScreen Solvinert Deep Well filter plate? | The extractables level of the MultiScreen Solvinert Deep Well plates is extremely low, making this product ideal for processing samples to be analyzed by HPLC or LC/MS/MS. |
| When I wet out the membranes in the MultiScreen Solvinert Deep Well plate they become translucent. Is this normal? | Yes, this is completely normal and expected. |
| Is the MultiScreen Solvinert Deep Well plate automation compatible? | Yes, the MultiScreen Solvinert Deep Well plate is automation compatible. |
| When using the MultiScreen Solvinert or Solvinert Deep Well for Total Drug Analysis, can I add the serum all at once to each well? Why do I have to drip the serum in slowly for the manual procedure? | Slow addition of the serum to each well allows for maximum surface area exposure of the plasma with the acetonitrile. Subsequent mixing of the solution provides for complete dispersion of the plasma into the acetonitrile thus allowing complete protein precipitation. |
| When using the MultiScreen Solvinert or Solvinert Deep Well for Total Drug Analysis, why does the protocol require 90% acetonitrile? Can I use 100% acetonitrile? | The specific density of acetonitrile and plasma are rather different, which can lead to incomplete mixing. Prewetting of the acetonitriile with 10% water improves mixing and allows for complete integration of the plasma/serum into the acetonitrile. |
| When using the MultiScreen Solvinert or Solvinert Deep Well plate for Total Drug Analysis, how fast should I shake the plate after adding the serum/plasma and acetonitrile to each well? At what setting should my plate shaker be set? | It is recommended to shake the plate as fast as possible without spilling the solution from the wells. With the clear wells of the Solvinert Deep Well plate it is easy to see the level of the solution in the wells as it shakes. Simply turn the dial of the plate shaker up until the solution is approaching the top of the wells. It has been our experience that plate shakers can have different gauge settings, and even the same setting on different plates can have different actual rates. |
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Kategorie
| Life Science Research > Cell Culture and Systems > Cell Culture Flasks, Plates, & Slides > Multiwell Plates |
Determining the concentration of drug in plasma or serum at various time points after administration is necessary to calculate the pharmacokinetics (PK) of a drug. PK, in turn, is an important component in the absorption, distribution, metabolism, and excretion (ADME) profile. Precise knowledge of ADME properties enables accurate determination of the proper drug dosage to maintain therapeutic drug levels without risking toxicity.
MultiScreen Deep Well Solvinert and MultiScreen Solvinert filter plates are pre-validated for in-plate precipitation of proteins from plasma or serum for total drug analysis. The plates enable fast, discreet and complete filtrate transfer to provide an automation-compatible platform for sample preparation prior to total drug analysis. Filtration with the Solvinert family of filter plates provides protein-free filtrate that is compatible with HPLC-MS or HPLC-UV analysis.
The MultiScreen Deep Well and MultiScreen Solvinert filter plates are robust and reliable platforms that generate reproducible results. Samples isolated from serum by protein precipitation and filtration through these plates are essentially protein-free and show high compound recovery and no interference from extractables.
Automation-compatible MultiScreen Deep Well Solvinert Filter Plates Yield Reproducible Results for Total Drug Determination
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Warfarin was spiked into bovine serum followed by protein precipitation and filtration through MultiScreen Deep Well Solvinert filter plates. The assay was carried out manually and with complete automation. Drug concentrations in the filtrates were analyzed by LC-MS/MS and are compared. The linearity of results for both automated and manual methods (R2 values= .999) demonstrates that drug samples in serum can be reliably prepared manually or automated by precipitation and filtration in a MultiScreen Deep Well Solvinert filter plate. The high degree of linearity indicates that filtrates generated following this procedure allow accurate LC-MS/MS analysis across a wide concentration range.
High Performance Plate Designs
Automation-compatible MultiScreen Solvinert filter plates are available in 96-well deep well (2 mL) and standard (500 µL) formats. The one-piece plate designs incorporate rigid sidewalls to facilitate use with robotic gripper arms and provide space for barcode labels.
All MultiScreen Solvinert plate materials are selected for broad chemical compatibility and enable extended in-plate incubations with no leaking.
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Optimized for Drug Discovery and HPLC Sample Preparation
MultiScreen Solvinert filter plates – both deep well and standard – are optimized for drug discovery applications including total drug analysis, NCE cleavage from solid phase libraries, and sample preparation prior to HPLC or LC/MS/MS. The plates and membranes demonstrate low binding, low extractables and high recoveries. For chromatography separations, the MultiScreen Column Loader accessory can be used to create 96 minicolumns per plate.
MultiScreen Deep Well with hydrophobic membrane and prefilter is also well suited for total drug analysis in plasma by protein precipitation.
Hydrophilic and Hydrophobic PTFE Membranes Available
MultiScreen Solvinert filter plates are available with chemically-resistant hydrophilic and hydrophobic membranes to accommodate aqueous and non-aqueous samples. MultiScreen Deep Well Solvinert filter plates are also available with an optional pre-filter to use with highly particulate media. Both membranes are highly retentive (>99% retention of acid precipitated BSA).
For the broadest chemical resistance, hydrophobic PTFE membrane is recommended. Designed for extended sample incubation times and the lowest extractables, hydrophobic PTFE membrane is ideal for NCE cleavage and cleanup. It is also suitable for in-plate protein precipitation and sample recovery following in-plate compound cleavage from solids or beads. The plates are also used for peptide synthesis.
Hydrophilic PTFE membrane is optimized for low drug and protein binding with excellent throughput in typical aqueous and solvent sample preparation. High sample recoveries and low extractables provide for optimum analysis by HPLC and LC/MS/MS. The membrane is suitable for applications including natural product screening, aqueous solubility testing and total drug analysis.
Performance
High Recovery of Drug in Solvent-based Preparations
)
90%) for both hydrophilic and hydrophobic MultiScreen Solvinert plates. (The sample was pre-precipitated by the addition of CACN followed by vigorous mixing.) Equivalent results are seen with MultiScreen Deep Well Solvinert filter plates." />
Seven drugs were tested for percent recovery by acetonitrile plasma precipitation. Plasma stock samples (5 mL) were spiked with drug (100 µM stock and tritiated) to a final 5 µM drug concentration followed by 1-hour incubation. Protein was precipitated by the addition of acetonitrile (15 mL) and the solution was vortexed vigorously. For each drug a 300 µL aliquot of the supernatant was added to 8 wells per plate. The samples were filtered by vacuum filtration (12" Hg) and filtrates were collected. Percent recovery was determined by comparing an aliquot (100 µL) of filtrate to an aliquot of the precipitated stock (cpm filtrate/cpm precipitated stock). Results show superior drug recovery (>90%) for both hydrophilic and hydrophobic MultiScreen Solvinert plates. (The sample was pre-precipitated by the addition of CACN followed by vigorous mixing.) Equivalent results are seen with MultiScreen Deep Well Solvinert filter plates.
Note: The storage lid does not have the same solvent resistance properties as the filter plate and can be damaged by exposure to some solvent vapors.
*Dioxane, Hexane and Benzene are not recommended for use in Deep Well plates
The data presented in this chart are a compilation of testing by Millipore with certain chemicals and manufacturers' compatibility recommendations. These data are intended to provide expected results when filtration devices are exposed to chemicals for 48 hours at 25 °C (77 °F), unless otherwise noted.
| Extended Incubation Times with No Leaking | ||
| Solution | 24-hour Incubation Performance | |
| Hydrophilic PTFE Membrane |
Hydrophobic | |
| Milli-Q 18 MΩ Water | R | R |
| EtOH, 100% | R | R |
| MeOH, 100% | R | R |
| MeCl2 | NR | R |
| DMSO/PBS (5%/95% v/v) | R | R |
| ACN, 100% | R | R |
| ACN/H2O (75%/25% v/v) | R | R |
| TFA/ACN (80%/20% v/v) | NR | R |
| DMSO, 100% | R | R |
| DMF, 100% | R | R |
| NaOH, 1.75N | R | R |
| R = Yes NR = Not recommended. Some wells exhibited partial or complete drip out in 24 hour testing. May be compatible for shorter incubation times. |
||
Table 2. MultiScreen Solvinert Filter Plates are designed to support extended incubations. This solvent compatibility table shows results reported for 200 µL of liquid with 24 hours incubation at room temperature.
| Filter Plates | MultiScreen Deep Well Solvinert | MultiScreen Solvinert |
| Sample Volume per Well | ||
| Recommended, mL | 50 µL - 1.8 mL | 50 µL - 0.45 mL |
| Maximum, mL | 1.9 mL | 0.5 mL |
| Filtration Parameters | ||
| Relative Centrifugal Force | Maximum 3,000 x g | Maximum 3,000 x g |
| Maximum Vacuum, mm Hg (in. Hg) | 610 (24) | 610 (24) |
| Materials | ||
| Base Plate | Polyolefin copolymer | Polyolefin copolymer |
| Membrane | Hydrophilic Polytetrafluoroethylene (PTFE) | Hydrophilic Polytetrafluoroethylene (PTFE) |
| or | Hydrophobic Polytetrafluoroethylene (PTFE) | Hydrophobic Polytetrafluoroethylene (PTFE) |
| Prefilter | Polypropylene (optional) | N/A |
| Dimensions | ||
| Membrane Area | 0.28 cm² | 0.28 cm² |
| Plate Length, mm | 128 mm | 128 mm |
| Plate Width, mm | 85.5 mm | 85.5 mm |
| Plate Depth, mm | 40.7 mm | 14.6 mm |