Part No: AN795Issued year: 2013File size: 1.09mbFile type: pdf
This application note describes a method of extraction for metanephrine and normetanephrine from synthetic
urine using EVOLUTE® EXPRESS WCX 96-well SPE plates.
A new solid phase extraction technology has been developed to reduce
the time associated with sample preparation methods by
eliminating steps in the extraction workflow. Leveraging this
technology, a method protocol was developed using a load,
wash and elute approach. This method was optimized for
Part No: AN080Issued year: 2013File size: 0.26mbFile type: pdf
This application note describes a method for sample preparation
of different matrixes prior to amino acid analysis according to IS
ISO 13903:2005 or the similar AOAC Official Method 994.12 using
Biotage® Initiator+ to speed up the process.
Part No: PPS436.v1Issued year: 2016File size: 0.17mbFile type: pdf
Sanford Burnham Prebys Case Study.
Fewer transfers and the ability to evaporate difficult solvents right in
the scintillation vial has made Biotage® V-10 the go-to evaporator at
Sanford Burnham Prebys (SBP), a top medical research institute.
Part No: TN-0012.0806Issued year: 2008File size: 0.09mbFile type: pdf
Biotage FLASH 400™ Production-Scale System The FLASH 400 is a complete skid-mounted system designed for large-scale flash chromatography and adsorption purification. The FLASH 400 uses prepacked cartridges and radial compression for performance and reliability. Built with industrial-grade components that are appropriate for operations under FDA regulations and cGMP standards, the FLASH 400 is rapidly becoming the first choice of pharmaceutical and contract manufacturing companies around the world for critical purification applications.
Part No: AN059Issued year: 2012File size: 0.63mbFile type: pdf
Flash chromatography is the primary technology for separating, purifying, and isolating both synthetic organic compounds and natural products. Until recently, automated flash purification systems were limited to user-selected detection and collection wavelengths to fractionate separated compounds from sample mixtures. In this application note we demonstrate how the advances in Isolera Spektra are used in the purification of a spinach extract.
Part No: AN060Issued year: 2012File size: 1.02mbFile type: pdf
Isolera Spektra provides definitive fraction purity assessment
in both 2-D and 3-D prior to any post flash analysis. With this
information chemists quickly know if a fraction contains a pure
product suitable for further mass and structure confirmation
or whether the impure fractions must be re-purified. This
knowledge improves synthesis throughput and avoids any
embarrassment of submitting impure fractions for analytical
Part No: AN092.v1Issued year: 2014File size: 1.12mbFile type: pdf
The UV absorption spectrum of some solvents
overlaps with the product they dissolve, meaning
that fraction collection processes cannot distinguish
between solvent and product. Luckily, there is
technology that solves this problem.
Flash purification is a separation technique developed in 1978 by Professor W.C. Still that uses a stationary phase (a column or cartridge filled with an insoluble solid support) and a mobile phase (elution solvent mixture) to separate and purify a mixture of organic compounds.
Part No: AN095Issued year: 2014File size: 0.65mbFile type: pdf
Elution of polar molecules in flash chromatography sometimes requires solvent additives, making purification more cumbersome. The Isolera™ family of flash purification instruments and Biotage® SNAP KP-NH cartridges significantly reduce the extra work involved.
Part No: TN-0006.0806Issued year: 2006File size: 0.86mbFile type: pdf
Purify up to 400 grams of compound at 1 L/min with the Biotage Flash 150™ system, up to 2x faster than traditional glass columns. This robust stainless steel system safely operates at 100 psi enabling high flow rates and the use of higher viscosity solvents. A variety of cartridge media provides
chemists with selectivity choices for optimal purification conditions. Simple and reliable, this system contains everything needed to begin your separations.
Part No: TN407.1107Issued year: 2007File size: 0.07mbFile type: pdf
FlashVac-10 and -20 Sample Processing Manifolds are used
to process standard Luer tipped ISOLUTE work-up columns.
Constructed from glass or high density polyethylene, the
FlashVac manifolds are compatible with commonly used
reaction and work-up solvents.
Part No: AN086.v2Issued year: 2014File size: 1.02mbFile type: pdf
A novel resin from MIP Technologies, RENSA® PY, is able to fractionate colored compounds present in beetroot juice using mild and purely aqueous elution conditions without the use of salts or organic solvents.
Part No: Issued year: 2015File size: 1.88mbFile type: pdf
User Report: Isolera™ Dalton, Okayama University. The laboratory of Prof. Hiroyuki Miyachi at Okayama University installed Biotage’s Isolera Dalton, an automated mass-directed purification system for flash chromatography, allowing the fractionation and collection of optical isomers. “I felt like the world’s first mass-detection automated purification system had finally arrived.”
Part No: AN792Issued year: 2013File size: 2.3mbFile type: pdf
This application note describes fully automated liquid handling of the extraction of a range of drugs from urine, which are typically screened for forensic toxicology panels, using ISOLUTE® SLE+ 96-well supported liquid extraction plates.
drugs of abuse, TECAN, automated, ISOLUTE SLE+
Part No: P150Issued year: 2016File size: 0.39mbFile type: pdf
Interest in peptide-based therapeutics has risen dramatically over the last decade. More specifically, significant energy has been dedicated to cyclic peptides, their synthesis, their biological activity, and their biological stability as potential alternatives to their linear counterparts. However, synthesizing a cyclic peptide creates significant challenges not encountered in linear peptide synthesis.
Herein we present a synthesis of oxytocin describing optimization for fully automated, on-resin disulfide bond formation utilizing Branches™, a unique feature enabling visualization and specific programing for cyclic and branched peptide synthesis. We will highlight the transferability of this optimized synthesis to larger scale syntheses and conclude with the fully automated synthesis of cyclized oxytocin incorporating an isotopically labeled Leu residue, enabling use as an internal standard in MS experiments.
Part No: TN113Issued year: 2006File size: 0.09mbFile type: pdf
ISOLUTE Confirm HCX columns are based on cation exchange and C8 mixed mode chemistries. Basic drugs are therefore retained by two primary retention mechanisms - ionic and non-polar. This allows a more rigorous interference elution regime to be used, leading to a very clean final extract, as many non-polar interferences which are retained by a non-polar interaction alone, can be eluted selectively, prior to elution of the drug.
Part No: P024Issued year: 2008File size: 1.9mbFile type: pdf
It is well known that traditional liquid-liquid extraction (LLE) provides very clean extracts prior to LC/MS analysis. Supported liquid extraction is analogous to traditional LLE, however, analyte partitioning takes place using an inert support material, rather than two immiscible liquids. This provides excellent extraction efficiencies while alleviating many of the tedious liquid handling issues associated with LLE.
Part No: TN116Issued year: 2006File size: 0.07mbFile type: pdf
The extraction of basic drugs from biological fluids using a purely non-polar retention mechanism (e.g. C4, C8 or C18) can lead to extracts that contain a large amount of non-polar co-extracted material that can interfere with subsequent analysis. Conversely, extraction mechanisms based on ion exchange interactions can be non-robust due to the variable ionic strength of the sample matrix.
Part No: TN129Issued year: 2005File size: 0.07mbFile type: pdf
The use of ISOLUTE HCX mixed-mode sorbents is widely accepted for providing high purity extracts of basic drugs from biological fluids. The ISOLUTE HCX-Q sorbent utilizes a combination of weak cation exchange and C8 non-polar retention mechanisms.
Part No: Issued year: 2014File size: 1.02mbFile type: pdf
Testing for drugs of abuse in oral fluids can strongly benefit the criminal justice field as a less invasive and cost-effective approach for drug detection when compared to blood or urine sampling. Oral fluid analysis has facilitated laboratory analysis for many drugs of abuse and is a constantly evolving analysis procedure which benefits from increasingly sensitive methods of detection. The Laser Diode Thermal Desorption (LDTD) source combined with Mass spectrometry is presented as a new screening tool for drug analysis in Oral Fluids. Analysis speed of LDTD provides accurate results in seconds in combination with exceptional specificity of MS instruments make a powerful platform for the screening of different drugs of abuse and new emerging drugs. Different extraction procedures are available; however those methods depend on specific drug conditions: basic or acid drugs / hydrophilic or hydrophobic. A new extraction approach, Supported Liquid Extraction (SLE+) is evaluated as generic extraction procedure.
Part No: AN056Issued year: 2012File size: 0.2mbFile type: pdf
Quinolines are an important class of broad-spectrum antibiotics1 that were traditionally obtained by refluxing an aniline and diethyl ethoxymethylemalonate (Scheme 1) for several hours often in low yield.2 Heating the reaction mixture by microwaves to temperatures above the boiling points, with or without solvents, improves the yields and shortens the reaction time dramatically.3-6
Part No: AN106Issued year: 2015File size: 0.3mbFile type: pdf
The term “Green Chemistry” has become a major part of the
science community’s lexicon. In this application note we will
look at two areas for flash chromatography:
1. Replacing chlorinated solvents with those considered more
2. Reducing solvent use and waste generation with more
thoughtfully applied chromatography principles.
Part No: P158Issued year: 2017File size: 0.27mbFile type: pdf
As reversed-phase flash chromatography gains traction in
medicinal chemistry labs the need to monitor its cost and
safety are becoming more important. Commonly used
reversed-phase solvents typically include water with an
organic solvent such as methanol or acetonitrile – each
have advantages and disadvantages.
Part No: AN801Issued year: 2003File size: 0.27mbFile type: pdf
The heat flow parameter can be used to monitor the progress of a reaction, and can be useful
in determining reaction rates. A heat flow probe study was executed on the Advantage
Series™ 3400 process chemistry workstation for the hydrolysis of acetic anhydride in water
at temperatures from 25 to 55 °C. The observed rate constant for the hydrolysis was determined
from the heat flow measurements. An Eyring plot was produced and the enthalpy and
entropy of activation were determined. Finally, the heat of reaction was measured based on
the total amount of heat that was observed during the hydrolysis.
Part No: AN036Issued year: 2001File size: 0.55mbFile type: pdf
Activated carbon is an adsorbent media used to remove colored compounds from solution. Using this media in the Biotage FLASH-AC cartridge as a packed bed improves adsorbent performance. Increasing the temperature of the solvent modifies adsorption and alters performance of the decolorization process. Elevated temperatures increase product solubility, therefore requiring less solvent than at room temperature. The increased concentration of product in a hot solution also simplifies crystallization.
Part No: AN040Issued year: 2003File size: 0.4mbFile type: pdf
Research in CNS drugs is primarily centered on nitrogen heterocycle chemistry. Basic amines are difficult to purify using traditional silica chromatography because of strong interactions between acidic silica and the molecules’ basic amine groups. These interactions cause band spreading and poor compound recovery. Solutions employed to counteract this phenomena include adding a competing amine (e.g. triethyl amine or ammonium hydroxide) to the flash chromatography solvent system or using reversed-phase HPLC with a buffered solvent system.
Part No: P172Issued year: 2017File size: 0.49mbFile type: pdf
We investigate the use of reversed-phase high performance flash chromatography (HPFC) to quickly purify large quantities of crude synthetic peptides.
(6th Solid Phase Peptide Synthesis Symposium & 12th Australian Peptide Conference, Queensland, Australia, Oct. 2017)
Part No: Issued year: 2018File size: 0.66mbFile type: pdf
This poster presents a high sensitivity method for extraction and analysis of estrogens in human plasma. Sample preparation utilizes Biotage ISOLUTE SLE+ plates, providing high analyte recovery and low matrix effects.
MSACL 2018, Palm Spings, CA
Part No: Issued year: 2014File size: 0.34mbFile type: pdf
In this poster, an IONICS 3Q 220 triple quadrupole mass spectrometer using a load, wash, elute method protocol with recently advanced solid phase extraction (SPE) sorbent technology (EVOLUTE EXPRESS WCX) was used to perform analysis of the two compounds. As a result low levels of metanephrine and normetanephrine are detectable in plasma with short sample preparation and LC run times. This LC-MS/MS method provides a fast, sensitive, accurate, and reproducible solution for the analysis of ME and NME in plasma.
Part No: AN303Issued year: 2007File size: 0.14mbFile type: pdf
Heck reactions utilizing a thermostable catalyst in combination with microwave irradiation have been performed. A substantial enhancement of reaction rates as well as excellent regiocontrol could be obtained under microwave conditions without using inert gas. A slight increase in yield was also noted.
Part No: CM-HMPB-0810Issued year: 2010File size: 0.29mbFile type: pdf
For peptides acid, we recommend using the HMPB-ChemMatrix
as this resin will provide high crude purity and a recovery yield of
90-95%. The Wang-ChemMatrix will produce similar crude
peptide purity, but the recovery yield is lower (60-70%). On
request, we can provide preloaded HMPB-ChemMatrix resins with
all 20 naturally occurring amino acids. Other amino acids can be
preloaded on special request.
Part No: AN747Issued year: 2011File size: 0.18mbFile type: pdf
Biotage® PRESSURE+ manifolds deliver positive pressure, parallel processing for 96 well plates and 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. ISOLUTE SLE+ Supported Liquid Extraction plates and columns offer an efficient alternative to traditional liquid-liquid extraction (LLE) for bioanalytical sample preparation, providing high analyte recoveries, no emulsion formation and greatly matrix effects associated with biofluid analysis.
SLE+, SLE, Supported Liquid Extraction, positive pressure manifold, Pressure+96, Pressure+48
Part No: UI333Issued year: 2015File size: 0.89mbFile type: pdf
If high back pressure is detected in the makeup pump lines, and/or the High Makeup Flow Pressure dialog appears, the microfilter may be partially clogged and needs replacing. This is a step by step guide.
Part No: UI319.v3Issued year: 2017File size: 0.7mbFile type: pdf
Biotage®SNAP Ultra cartridges are compatible with CombiFlash® Rf Systems. As a convenience, Biotage SNAP Ultra cartridges with Luer slip connectors are available. Contact your Biotage representative for ordering information.
Part No: AN078Issued year: 2013File size: 1.05mbFile type: pdf
In this application note we demonstrate the power of mass detection in Isolera™ Dalton by investigating the incomplete separation of two dyes, Butter yellow (Mass 225.29 g/mol) and Sudan red (Mass 278.31 g/mol), by normal phase flash purification.
Part No: P053Issued year: 2013File size: 1.22mbFile type: pdf
Immunosuppressant drugs are extremely important for therapeutic drug monitoring (TDM) regimes, requiring robust and reliable methods for their analysis. This poster aims to demonstrate strategies for the extraction of various immunosuppressant drugs from whole blood providing clean samples prior to UPLC-MS/MS analysis. Spiked whole blood was extracted using a variety of protocols to investigate optimal combination of drug recovery and extract cleanliness. UPLC-MS/MS was conducted using a Waters ACQUITY UPLC coupled to a Quattro Premier XE triple quadrupole mass spectrometer. Acceptable extraction recoveries were obtained showing excellent extract cleanliness, demonstrating a reliable method for mass spectrometric approaches for these analytes.
Immunosuppressants, SLE, Supported Liquid Extraction, ISOLUTE, MSACL, San Diego, 2013
Part No: P140Issued year: 2016File size: 0.31mbFile type: pdf
Reversed-phase chromatography is typically used when you need to separate several milligrams of relatively polar compounds that either are not soluble in normal-phase solvents or are not compatible with bare silica
because they react, stick, or both. If you are currently using reversed-phase at preparative scale, such as flash chromatography, you know the mobile phase limitations – water with either methanol, acetonitrile, or
THF. As with normal-phase flash chromatography, when it comes time to purify you want your crude sample fully solubilized in the weakest possible solvent at the highest possible concentration.
Part No: AN075.v2Issued year: 2013File size: 0.49mbFile type: pdf
In this application note we present a new type of polymeric reverse phase adsorbent that is able to provide improved separations of a range of small chemical compounds and also peptides evaluated side by side to a comparable benchmark DVB-Styrene macroporous resin.
MIP, molecularly imprint resins
Part No: AN063Issued year: 2012File size: 0.48mbFile type: pdf
Pyrazines are a class of organic molecules often used to provide flavor to foods. They are typically synthesized but some are found in fruits and vegetables, e.g. grapes, bell peppers, peas, asparagus, beetroot, tobacco, and roasted foods. Pyrazine’s heterocyclic chemistry can yield some challenges to their purification due to the various separation kinetics between the compound and silica. Biotage SNAP Ultra.