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: 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.
Part No: P160Issued year: 2017File size: 0.24mbFile type: pdf
Although capable of very high resolution, RP-HPLC is often limited by low column loading capacity, therefore demanding a significant time investment for peptide purification. As an alternative strategy, reversed-phase flash chromatography can also be used to purify synthetic peptides. The larger particle size used in flash column chromatography enables much larger loading capacity, thereby significantly reducing the time required for peptide purification.
Part No: PPS375.v1Issued year: 2015File size: 0.3mbFile type: pdf
User report: Flash instruments. Chugai Pharmaceutical uses Biotage flash chromatography products for drug discovery research. When deciding to convert from manual open-column procedures to automated systems, they chose successive generations of Biotage products, ranging from the Flash+® packed column to the Biotage® Horizon, SP1, Isolera™ Spektra, and Isolera™ Dalton automated flash chromatography systems.
Part No: AN041Issued year: 2001File size: 0.16mbFile type: pdf
Normal-phase flash purification is commonly used by organic chemists in pharmaceutical drug discovery and
process development labs. However, for many synthesized products (e.g. peptides, nucleotides and basic
drug candidates) purification on standard flash silica is not an option due to irreversible adsorption, chemical
interaction and/or solubility issues. Reversed-phase flash purification is an excellent solution for these applications. Yet, this technique has been used sparingly because of perceived lower loading capacity, higher
operating pressures and a scarcity of publications addressing reversed-phase flash chromatography.
Part No: AN039Issued year: 2002File size: 0.64mbFile type: pdf
For pilot and production scale purification drugs, the use of a large particle-size media is common. Improving the purification throughput is limited by the media’s large particle size. Such a case was recently encountered when a pharmaceutical company attempted to improve the efficiency of a large-scale purification. In their current process, the use of a 350-600 µm polystyrene-type resin resulted in a lengthy purification cycle and low separation efficiency. However, the labile properties of the targeted component required a shorter purification cycle and high purification.
Part No: TN128Issued year: 2006File size: 0.09mbFile type: pdf
This technical note describes the use of derivatization techniques to separate aliphatic
secondary amines from aliphatic tertiary amines in a mixture using the strong cation
exchange sorbent, ISOLUTE® SCX-2.
Part No: PPS385Issued year: 2015File size: 4.87mbFile type: pdf
Our carefully selected portfolio of industrial scale products have a proven track record of successful applications and use in scale-up projects. We can support the discovery, development and manufacturing of customer pharmaceutical and biotechnology products, from pre-clinical, phase I, II and III to small scale commercial operations.
Part No: TN-0028.1222Issued year: 2012File size: 1.6mbFile type: pdf
The Initiator+ is a flexible system that utilizes all Biotage vials, from 0.2 to 20 mL,
in any order or combination, at any time without system modifications, delivering greater flexibility and direct scale-up from milligrams to grams.
Part No: TN-0029.0611Issued year: 2011File size: 1.07mbFile type: pdf
In peptide synthesis mode, this is the perfect tool for chemists synthesizing peptides and peptidomimetics, including difficult modifications and labeling of sequences, by using single or multi-step procedures. The intuitive Initiator 4.0 software controls the instrument functions and by using the pre-defined methods, synthesis can be started immediately.
Part No: AN090Issued year: 2014File size: 1.95mbFile type: pdf
In mass detection, the presence of ions can suppress peaks
from the analytes of interest. This study concludes that
Isolera™ Dalton performs successful separations even with
ions present in the sample.
Part No: AN088.v1Issued year: 2014File size: 1.59mbFile type: pdf
Fragrance compounds, being invisible to UV, can be
elusive and difficult to isolate. In this application, a four
component mixture of fragrance compounds was purified
using Accelerated Chromatographic Isolation™ (ACI) and
identified by mass detection.
Part No: AN600Issued year: 2005File size: 0.08mbFile type: pdf
This Application Note reviews the use of ISOLUTE PPT+ plates for the isolation of drugs from biological fluids, including the method, recovery data and the impact of the filtrate’s cleanliness on analyte quantitation.