ISOLUTE® PLD+ Protein and Phospholipid Removal plates offer a substantial improvement in extract cleanliness compared to traditional protein precipitation techniques for bioanalytical sample preparation.
This application note describes a simple, effective ISOLUTE® PLD+ protocol for the extraction of 25-hydroxy vitamin D from serum, demonstrating high, reproducible analyte recoveries with low protein and phospholipid content in the extracts.
Tags: 25-OH Vitamin D2, 25-OH Vitamin D3, 25-hydroxy-vitamin D, 96-well plate, Application Notes, Clinical, English, LC-MS/MS, Serum, Vitamin D, Vitamins
This application note describes the use of the ISOLUTE® PLD+ Protein and Phospholipid Removal Plate for clean-up of a range of acidic, basic and neutral drugs from plasma. ISOLUTE® PLD+ Protein and Phospholipid Removal Plates are suitable for clean-up of a range of analytes with widely differing functionality and polarity characteristics from plasma, giving high recoveries, good reproducibility and excellent extract cleanliness.
Tags: 96-well plate, Acetaminophen, Acidic, Amitriptyline, Application Notes, Atenolol, Basic, Bretylium, Brompheniramine, Clinical, English, Fluoxetine, Forensic, Ketoprofen, LC-MS/MS, Metoprolol, Mianserin, NSAIDs, Naltrexone, Neutral, Pharma, Plasma, Procainamide, Quinidine, Ranitidine, Salbutamol, Sulindac, Tricyclic Antidepressant, p-toluamide, β-blockers
In this catalog product information is displayed with easy-to-follow application details and supporting documentation.
Ordering information is found following each product listing.
This poster demonstrates the use of a novel protein and phospholipid depletion plate, for the extraction of 25-hydroxy vitamin D from serum. The extraction protocol was ultimately transferred to an SPE automation platform and method performance versus manual processing was compared.
Tags: 25-OH Vitamin D2, 25-OH Vitamin D3, 25-hydroxy-vitamin D, 96-well plate, Biomarker, Clinical, English, Extrahera, LC-MS/MS, Posters, Serum, Vitamin D, Vitamins
This poster describes a comparison of 25-hydroxy Vitamin D Extraction using Supported Liquid Extraction and Phospholipid Depletion Plate Technology using Manual and Automated Sample Preparation prior to LC/MS Analysis. It was presented at MSACL 2015, San Diego
Tags: 25-OH Vitamin D2, 25-OH Vitamin D3, 25-hydroxy-vitamin D, 96-well plate, Clinical, English, LC-MS/MS, Posters, Serum, Vitamin D, Vitamins
This poster presents the ISOLUTE PLD+ Protein and Phospholipid removal plate, highlighting its ease of use and excellent performance with respect to sample clean up, analyte recovery and elimination of back pressure build up in the UPLC system.
Tags: 96-well plate, Acetaminophen, Acidic, Amitriptyline, Atenolol, Basic, Bretylium, Brompheniramine, Clinical, English, Fluoxetine, Forensic, Ketoprofen, LC-MS/MS, Metoprolol, Mianserin, Naltrexone, Neutral, Pharma, Pharmaceuticals, Plasma, Posters, Procainamide, Quinidine, Ranitidine, Salbutamol, Sulindac, p-toluamide
This poster evaluates the performance of a novel 96-well filter plate (ISOLUTE PLD+ Protein and Phospholipid Removal Plate) for
the simultaneous removal of proteins and phospholipids from serum samples prior to LC-MS/MS analysis.
Tags: 96-well plate, Acidic, Amitriptyline, Analgesics, Atenolol, Basic, Bretylium, Clinical, English, Fluoxetine, Forensic, Ketoprofen, LC-MS/MS, Metoprolol, Mianserin, NSAIDs, Naltrexone, Pharma, Plasma, Posters, Procainamide, Quinidine, Ranitidine, Salbutamol, Serum, Sulindac, Tricyclic Antidepressant, β-blockers
The new ISOLUTE® PLD+ Protein and Phospholipid Removal Plate is highlighted in this poster. Protein and phospholipid removal, and analyte recovery are included, along with data illustrating the positive impact of clean (PL and protein free) samples in maintaining analyte signal intensity and low UPLC column back pressure over multiple LCMS runs.
Tags: 96-well plate, Clinical, English, Forensic, LC-MS/MS, Pharma, Posters
ISOLUTE PLD+ plates remove >99 % of plasma proteins and phospholipids, the main causes of ion suppression, leading to cleaner extracts and increased sensitivity (signal-to-noise
(S/N)) for a broad range of analytes.
Tags: 96-well plate, Clinical, English, Forensic, LC-MS/MS, Pharma, Product Notes
A poster from Biotage labs presented at ASMS 2015 demonstrates the applicability of ISOLUTE® PLD+ for the extraction of 25-hydroxy vitamin D from 100 or 150 μL of serum. This poster...12 May 2015
This application note describes the extraction of vitamin D metabolites from serum using ISOLUTE® PLD+ protein and phospholipid removal plates. This simple clean up technique provides an altern...20 March 2015
Effortlessly enhance analyte sensitivity with ISOLUTE® PLD+ protein and phospholipid removal plates from Biotage. In this video, product manager Claire Desbrow explains how and why they work so...13 November 2014
This application note describes the use of the ISOLUTE® PLD+ Protein and Phospholipid Removal Plate for clean-up of a range of acidic, basic and neutral drugs from plasma. The protein ...11 November 2014
ISOLUTE® PLD+ plates provide extremely efficient removal of proteins and phospholipids from plasma samples using a simple 3 step procedure. Recommended operating procedure fo...10 July 2014
Eight new posters by Biotage Scientists and collaborating groups have been presented at ASMS 2014, Baltimore, MD, USA. P81 A Reduced Workflow Method For The Extraction Of Vitamin B7 (bi...20 May 2014
Four new posters by Biotage R&D and Application Scientists have been presented this week at HPLC 2014, New Orleans, LA, USA. The studies focus on clean-up of biofluids using different sa...23 January 2014
Uppsala, Sweden, 22nd January, 2014 – Biotage (STO: BIOT), a leading global supplier of solutions and technology for analytical, medicinal and peptide chemistry, is pleased to announce the la...
What is the advantage of a phospholipid removal plate over the ISOLUTE SLE+ plate?
There are a number of technical considerations, but one of the key things is that the extracts can be injected direct into the LC-MS system, no evaporation-recon required. Elimination of this time consuming step is an important factor for many chemists.Browse more questions and answers >
How do ISOLUTE® PLD+ Protein and Phospholipid Removal Plates improve my sample preparation?
ISOLUTE® PLD+ Protein and Phospholipid Removal Plates combine effective protein and phospholipid removal in a single product providing a very effective but extremely simple sample clean-up for LC-MS/MS analysis. Requiring next to no method development, ISOLUTE PLD+ can be integrated quickly and easily into routine workflow,increasing productivity and reducing instrument downtime. ISOLUTE PLD+ plates remove >99 % of plasma proteins and phospholipids, the main causes of ion suppression, leading to cleaner extracts and increased sensitivity (signal-to-noise (S/N)) for a broad range of analytes.Browse more questions and answers >
How do ISOLUTE® PLD+ plates work?
In ISOLUTE® PLD+ plates, optimized frit components are combined with a novel proprietary sorbent to provide simultaneous protein and phospholipid removal. The procedure is very simple:
An appropriate volume of crash solvent is dispensed into the wells, and the proprietary treated top frit prevents
solvent dripping. This allows for a more efficient ‘solvent first’ protein precipitation to occur when plasma samples are added.
An appropriate volume of plasma or other blood based matrix sample is added to the crash solvent. To ensure efficient protein precipitation, sample should be added directly into the crash solvent with force, and plates left for 5 minutes for protein precipitation to complete. A depth filter prevents precipitated proteins from blocking or plugging the wells. Alternatively, ISOLUTE PLD+ plates can be used with vortex mixing or aspirate/dispense protocols.
Low vacuum or positive pressure is applied to the plate, drawing the sample through the proprietary sorbent bed, for collection of purified extract in a pre-positioned collection plate. The optimized bottom frit controls flow rates to ensure sufficient contact time for efficient phospholipid removal by the ISOLUTE PLD+ sorbent.Browse more questions and answers >
What is the ISOLUTE® PLD+ sorbent?
The ISOLUTE® PLD+ sorbent is a novel, proprietary multifunctional phase optimized to selectively retain phospholipids, but allow small molecule analytes (e.g. drugs and metabolites) with a broad range of chemical characteristics to pass through un-retained. Unlike some competitor products, phospholipid retention is not based on a purely hydrophobic retention mechanism.
The combination of efficient protein and phospholipid removal with high analyte recoveries leads to better sensitivity than other commercially available phospholipid removal products.
What analytes can be extracted using ISOLUTE® PLD+ plates?
ISOLUTE PLD+ plates can be used to extract wide range of acidic, basic and neutral analytes, with widely differing polarity/hydrophobicity. In most cases, a single simple generic methodology can be used, virtually eliminating method development time.Browse more questions and answers >
What detection limits should I expect when using ISOLUTE PLD+ plates for my sample preperation?
This is highly dependent on the analyte and analytical system employed, but increased sensitivity can be achieved using ISOLUTE® PLD+ plates compared with competitor products.Browse more questions and answers >
Can I add sample matrix first rather than crash solvent first when using ISOLUTE PLD+ plates?
Yes, but we have found that a better protein precipitation (crash), and hence better sample purification, is achieved using the solvent first approach. The optimized top frit in ISOLUTE PLD+ plates can hold up acetonitrile for up to 24 hours, so there is no danger of the crash solvent dripping through before all samples have been loaded onto the plate.Browse more questions and answers >
What crash solvent should I use with ISOLUTE® PLD+ plates?
We recommend acetonitrile, but methanol can be used as an alternative. Addition of 1 % formic acid to the crash solvent can lead to more efficient protein precipitation, and may increase recovery of certain analytes. For more viscous samples (e.g. whole blood), a crash solvent: sample of 4:1 (v/v) or higher may be required.Browse more questions and answers >
What vacuum or pressure levels should I use to process ISOLUTE® PLD+ plates?
We recommend the use of vacuum (-0.2 bar) or low pressure (3 psi) for processing ISOLUTE PLD+ plates. Using these gentle processing conditions, processing of samples should be complete in 5 minutes or less. Due to the optimized depth filter frit arrangement and narrow sorbent particle size range, ISOLUTE PLD+ are less prone to plugging/blockage than competitor products, so do not require the high pressure/vacuum conditions recommended by other vendors. As a consequence ISOLUTE PLD+ plates are easier to use than some competitor products on automation systems utilizing vacuum processing. For particularly viscous samples, it may be necessary to increase processing pressure/vacuum. In this case, phospholipids will be removed, but removal efficiency may be reduced.Browse more questions and answers >
How much sample can I load onto ISOLUTE PLD+ plates?
Most commonly, 100 μL of sample is loaded into 300–400 μL of crash solvent. However, 200 μL sample matrix with 600–800 μL of crash solvent can be loaded without loss of performance.Browse more questions and answers >
What sample volume should I get back using ISOLUTE® PLD+ plates?
As with all protein precipitation based methods, a loss of sample volume can be expected. This can vary with sample type and the solvent used. Typically, ~ 75% of the original sample plus crash solvent volume is recovered following purification.Browse more questions and answers >
What if I don’t have enough samples to utilize an entire ISOLUTE PLD+ 96-well plate?
You can process a partial ISOLUTE® PLD+ plate by sealing the unused wells with a piercable sealing cap (P/N 121-5204) or sealing tape.Browse more questions and answers >
What’s the maximum total volume (sample + crash solvent) that I can load into each well of an ISOLUTE® PLD+ plate?
We would not recommend exceeding a total volume of ~1500 μL, to prevent well to well cross contamination on processing. If vortex mixing, this volume should be reduced to a maximum of ~1350 μL.Browse more questions and answers >
Can I purify samples other than plasma using ISOLUTE PLD+ plates?
Yes, ISOLUTE PLD+ plates can be used to remove phospholipids from other sample matrices, including serum, whole blood and tissue homogenates.
Special considerations for whole blood
It is particularly important to ensure an efficient crash to prevent breakthrough of red blood cells/extract coloration.
Consider vortex mixing or aspirate/dispense cycles to improve crash efficiency.
Special considerations for tissue homogenates
Centrifugation of the sample prior to clean up on ISOLUTE PLD+
plates may improve performance.
How do I maximize analyte recoveries when using ISOLUTE PLD+ plates?
What should I do if I get a cloudy extract when using ISOLUTE PLD+ plates?
A cloudy extract suggests an incomplete protein precipitation has occurred, due to insufficient mixing of the sample and crash solvent. To ensure good mixing, try the following practical tips.
What if my analyte or sample type or volume is incompatible with ISOLUTE® PLD+ purification?
If your analytes or sample size requirements are not amenable to clean up using ISOLUTE PLD+ plates, we recommend evaluation of an alternative sample preparation technique such as supported liquid extraction (using ISOLUTE SLE+ plates or columns) or solid phase extraction (using EVOLUTE® or EVOLUTE EXPRESS SPE products).Browse more questions and answers >
Biotage® PRESSURE+ manifolds deliver positive pressure, parallel processing for 96 well plates, 1 mL 3 mL and 6 mL column formats. The systems utilize a consistent, uniform flow of positive pressure to move both low and high viscosity liquids through SPE plates and columns.
Biotage® VacMaster™-96 sample processing manifold has been designed with the needs of the bioanalytical scientist in mind. This manifold is ideal when processing 96-well SPE, supported liquid extraction or protein precipitation plates either manually or with an automated liquid handling system.
|Part No.||Description||Product type||Pack Size||Price|
|121-5202||Collection plate, 1 mL Square||Consumables||50||Login for price|
|121-5203||Collection plate, 2 mL Square||Consumables||50||Login for price|
|121-5213||Collection plate, 2 mL Round||Consumables||50||Login for price|
|918-0050-P01||ISOLUTE® PLD+ Protein and Phospholipid Removal Plate||Consumables||1||Login for price|
|Part No.||Description||Product type||Pack Size||Price|
|121-5204||Piercable Sealing Cap||Accessories||50||Login for price|