|High throughput sample preparation for the quantitation of drug compounds in serum samples|
|MultiScreen Deep Well Solvinert Filter Plate: High throughput sample preparation for the quantitation of drug compounds in serum samples|
|MultiScreen Solvinert and MultiScreen Deep Well Solvinert Filter Plates; 96-well chemically compatible sample prep platforms|
|Automated Sample Preparation Method for High Throughput Total Drug Analysis by LC-MS/MS|
|Automation of ADME Applications|
|Poster: Rapid Sample Preparation Method for High Throughput Total Drug Analysis by LC-MS/MS|
|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 2006
|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 2005
|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).|
|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 use only a few wells at a time?||It is recommended that all wells of the plate be filled, preferably with similar solutions and volume. If only a portion of the 96 wells are to be used, the remaining wells should be sealed with a mat-cap or sealing tape. Please see the User Guide for specific information.|
|When using the MultiScreen Solvinert Deep Well plate for Total Drug Analysis, why is the maximum recommended volume for the manual method only 1 mL total volume for a 2 mL plate?||For most efficient mixing of the sample, the volume recommended allows for vigorous mixing without spilling from the individual wells.|
|When using the MultiScreen Solvinert Deep Well plate for Total Drug Analysis, can I filter by centrifugation using any centrifuge with a carrier plate?||A deep well filter plate assembled on top of a deep well collection block is not compatible with all centrifuge rotors and plate carriers. The carrier must have at least 86 mm of clearance in order to be compatible with the deep well filter plate/Greiner receiver block assembly.|
|Can I use any plate shaker with the MultiScreen Solvinert Deep Well filter plate when running the Total Drug Analysis protocol?||Any plate shaker is compatible with the MultiScreen Solvinert Deep Well filter plate as long as it meets the following two criteria: 1. It must be able to achieve a high shaking speed to induce vigorous mixing. 2. It must have a solid platform on which to place the plate. It is important to note that some plate shakers have a foam base which has the potential to make contact with the spouts in the underdrain. In some circumstances, this contact can initiate flow through the plate.|
|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, why do I have to leave the vacuum on for 30 seconds after the solution has completely filtered? Won't that cause the acetonitrile to evaporate?||It is recommended to leave the vacuum on for an additional 30 seconds to ensure complete and discrete filtrate transfer into the collection plate. Although the wells have been completely evacuated, the filtrate has not completely cleared the underdrain. An additional 30 seconds of vacuum allows for the discrete transfer of the filtrate.
Vacuum filtration results in minimal evaportaion of acetonitrile. With a vacuum time of 2 minutes, only 10-20 uL of acetonitrile are lost due to evaporation.
|When using the MultiScreen Solvinert or Solvinert Deep Well plate for Total Drug Analysis, what is the maximum total volume I can use?||The maximum total volume is dependent on the designated usage of the plate.
For out of plate protein precipitation, up to 1.9 mL can be transferred to each well without shaking. The maximum volume with shaking for the device is 1.5 mL to ensure the solution does not spill out of the wells. With such volumes one must be wary of the shaker settings and monitor the liquid level.
With in-plate protein precipitation following the protocol, 1 mL is the recommended total volume for manual sample handling while1.25 mL is the maximum volume for the automated procedure.
|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).|
|MultiScreen Deep Well Solvinert Filter Plate User Guide|
Centrifugal and Chromatography AccessoriesCombine the cost savings of bulk media with the convenience of 96-well filtration plates.
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MultiScreen®HTS Vacuum ManifoldFor vacuum filtration of 96 and 384 samples
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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
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.
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.
High Recovery of Drug in Solvent-based Preparations90%) 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|
|Milli-Q 18 MΩ Water||R||R|
|DMSO/PBS (5%/95% v/v)||R||R|
|ACN/H2O (75%/25% v/v)||R||R|
|TFA/ACN (80%/20% v/v)||NR||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|
|Relative Centrifugal Force||Maximum 3,000 x g||Maximum 3,000 x g|
|Maximum Vacuum, mm Hg (in. Hg)||610 (24)||610 (24)|
|Base Plate||Polyolefin copolymer||Polyolefin copolymer|
|Membrane||Hydrophilic Polytetrafluoroethylene (PTFE)||Hydrophilic Polytetrafluoroethylene (PTFE)|
|or||Hydrophobic Polytetrafluoroethylene (PTFE)||Hydrophobic Polytetrafluoroethylene (PTFE)|
|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|