8 RP Analysis of RNA Markers using a Bioinert Column
Today, oligonucleotides have become more and more important in genetic testing, research, and forensics. To date a variety of oligonucleotide-based approaches have been developed for different pharmaceutical applications. Therefore, clean and reliable oligonucleotide standards are necessary for various analytical methods. One of these standards are RNA markers which are RNA fragments of a certain size which are used in gel electrophoresis to estimate the size of other RNA fragments.
The electron-rich backbone of oligonucleotides can cause irreversible adsorption on metal parts. This is a major issue for reversed phase liquid chromatography analyses as standard equipment and hardware such as tubing, frits or the standard column-body is made from stainless steel. The oligonucleotides bind through ionic interactions with the positively charged metal oxide layer of the metallic surfaces. This effect is enhanced when working at low to neutral pH as metals become more electropositive.
To overcome these challenges bioinert systems and columns such as the recently introduced YMC-Accura Triart columns are beneficial. YMC-Accura Triart columns have a bioinert coating on all surfaces, including the frits, to prevent any unwanted ionic interactions.
This application note shows the separation of 7 RNA markers from 100–1000 bases using a YMC-Accura Triart Bio C4 column at pH 7 and 80°C. All 7 peaks are very well resolved and show excellent recoveries from the first injection. It shows 10 consecutive injections using the YMC-Accura Triart Bio C4 as well as the corresponding standard YMC-Triart Bio C4 stainless steel column. The bioinert YMC-Accura Triart Bio C4 column provides reproducible results. In contrast, the same separation using a standard stainless steel column requires pre-conditioning and provides much lower sensitivities even after 10 injections.
More information online:
ilmt.co/PL/w1Aj and
ilmt.co/PL/EL4q 59616pr@reply-direct.com
Ready for the Future of Rapid Trace PFAS Screening
The analysis of polyfluoroalkyl substances (PFAS) is a well-discussed topic in today’s environmental market. Due to concerns about the effect on human health and environmental risks, measurements of PFAS concentrations in a wide variety of matrices are conducted. These studies aim to get a better insight into the effects of PFAS substances.
To underline the importance of this subject, a proposal to restrict the use of per- and polyfluoroalkyl substances (PFAS) was submitted in January 2023 to the ECHA by several European countries. This proposal aims to ultimately ban the use of PFAS substances.
Traditional PFAS analytical methods are focused on the speciation of the individual PFAS components with expensive separation techniques. Currently, thousands of PFAS components have been produced and identified for monitoring, which makes the analyses for these components more and more complex.
Due to this, there is a growing request for a standardised, fast and reliable PFAS screening method to optimise the sample throughput and reduce resources. TE Instruments’ solution for PFAS screening is the Xprep C-IC, designed to determine the total amount of PFAS components in aqueous matrices at ultra-low trace levels.
The determination of AOF (Adsorbable Organic Fluorines) is based on the same principle as AOX (Adsorbable Organic Halogens) analyses: A generally accepted and standardised technique used for decades in environmental laboratories for the monitoring of the levels of organic halides (Chlorine/Bromine/Iodine) in aqueous matrices. Where a sample is combusted, conditioned, and quantified by means of coulometric titration. For the analyses of AOF, the quantification step is performed by the use of an Ion Chromatograph (IC).
TE Instruments developed a fully automated, extremely compact sample preparation system covering oxidative pyrohydrolytic combustion, fraction collection, and sample injection towards the IC. The Xprep C-IC can introduce samples both via optimised direct injection (liquids module) and conventional boat-inlet (boat module) into a horizontal furnace. This automated sample-prep solution reduces the complexity of sample transfer and significantly improves user convenience.
A unique feature of the Xprep C-IC setup is the optional usage of a ceramic insert in the furnace combustion tube. While standard AOX analysis focuses on standard halides, the analysis of PFAS is mainly aimed at the analysis of elemental Fluoride. This is where the power of the ceramic insert comes into its place. In a standard situation, Fluoride attacks quartz glass and may thus create a defective combustion tube over time. With the usage of the ceramic insert, a protective layer protects the furnace tube quartz material resulting in a far longer lifetime compared to a standard quartz glass setup.
More information online:
ilmt.co/PL/5qbm 59680pr@reply-direct.com
PEEK HPLC Column Hardware
Get bio-inert, metal free HPLC columns with MicroSolv’s expanded line of PEEK column hardware. Extruded from Victrex® PEEK this hardware the highest quality on the market. Graphite is added (20%) to the formulation to achieve a black surface with a sheen and no other chemical or physical treatment is done leaving these columns more biocompatible.
PEEK or titanium frits of 2 µm porosity are available. The inner bore of the columns are not treated in any way and the mirror like surface is achieved through our proprietary extrusion method.
Available column packing equipment includes: packing adapters, seals, slurry reservoirs, frits & end fittings for packing stations. Now at reduced prices.
More information online:
ilmt.co/PL/w1lE 59472pr@reply-direct.com
INTERNATIONAL LABMATE - APRIL 2023
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