8 Environmental considerations


UV-Vis: • Temperature control: Maintain a stable temperature during measurements. Temperature affects absorbance.


• Solvent choice: Different solvents can alter absorption spectra. Choose a solvent compatible with your sample.


Fluorescence:


• Avoid photobleaching: Limit exposure to intense light to prevent fluorophore degradation.


• Shield from ambient light: Fluorescence is sensitive; even room lighting can affect measurements.


Data analysis and interpretation


UV-Vis: • Baseline correction: Subtract baseline absorbance to remove noise. • Quantitative analysis: Use calibration curves to convert absorbance to concentration.


Fluorescence:


• Correct for inner filter effects: High concentrations absorb excitation light, affecting emission. Apply correction factors.


• FRET efficiency calculations: Understand FRET efficiency based on donor and acceptor fluorescence.


Conclusion


Individually and when combined, these molecular spectroscopy technologies enable scientists to advance our knowledge and understanding of biomolecules. Both UV-Vis and Fluorescence spectroscopy are simple, cost effective and powerful for quantification and characterisation and are well-established techniques. Researchers can use both techniques side by side and gain greater insight for biological research.


References


1: Agilent Technologies (n.d.). High-Throughput Fluorescence-Based mAb Aggregation Analysis Solution. Agilent.com. Retrieved May 15, 2024, from https://www.agilent.com/en/product/molecular-spectroscopy/fluorescence- spectroscopy/fluorescence-accessories/high-throughput-fluorescence-based-mab-aggregation-analysis-solution


2: Clark, J. (n.d.). The Beer-Lambert Law. Chem.Libretexts.org. Retrieved May 15, 2024, from https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/Supplemental_Modules_(Physical_and_ Theoretical_Chemistry)/Spectroscopy/Electronic_Spectroscopy/Electronic_Spectroscopy_Basics/The_Beer-Lambert_Law


About the author


Ursula Tems has worked at Agilent Technologies since 2011 in Marketing and Product Management. Ursula is part of the Molecular Spectroscopy business, working on product and portfolio management and strategic planning with a focus on the Pharma and Biopharma end markets. With this experience Ursula is involved in new product design, development, and commercialisation.


Read, Share and Comment on this Article, visit: www.labmate-online.com


LCMS system supports mass-based peptide purification The VERITY®


271 LCMS system from Gilson combines the power of High-Performance Liquid Chromatography (HPLC) with Mass Detection


(MS) for conditional fraction collection. Initial screening for peptide purification can be done using smaller columns for scouting, and then the LC optimised conditions can be scaled up, allowing researchers to target a specific peptide for purification. In addition, with UV-VIS detection, the optional single quad VERITY 1920 MS Detector with electrospray ionisation (ESI) or atmospheric pressure chemical ionisation (APCI) ion sources supports targeted fractions collection using selected ion monitoring (SIM) channels.


With a mass range of 10-2000 m/z, the VERITY 1920 MS helps you easily detect large molecules such as peptides and small proteins. Many biomolecules carry multiple charges with electrospray ionisation so that even proteins of several tens of kDa can be measured within the 2,000 m/z range. The method takes advantage of the software’s powerful conditional fraction collection feature, which simultaneously combines UVVIS signals and up to five MS channels to improve selectivity and purity.


The VERITY 271 LCMS system is the ideal solution for peptide purification and laboratories seeking effective MS detection to isolate pure compounds from synthetic and natural products from lab to pilot scale. It makes mass detection affordable and straightforward for any purification work.


LC Software, which provides real-time and post-run MS and UV spectral data. More information online: ilmt.co/PL/5BD0 and ilmt.co/PL/EVXJ


The VERITY 271 LCMS system can be used to perform mass-based purification of target peptides from a mixture. The system is controlled using TRILUTION®


62604pr@reply-direct.com Element selective gas chromatograph mass spectrometer


Shimadzu Corporation introduces ELEM-SPOT, an innovative element selective gas chromatograph mass spectrometer. This ground-breaking system, developed in collaboration with TotalEnergies and leading universities in France and Spain, represents a significant advancement in biofuel quality assessment. By selectively detecting oxygen- and nitrogen-containing compounds within raw materials, ELEM-SPOT empowers users to enhance production efficiency and ensure biofuel quality.


In response to the growing demand for carbon-neutral energy sources, research has intensified on second-generation biofuels derived from non-edible materials like sugarcane peels, stalks, algae, and waste plastics. However, the complex composition of these materials posed challenges in identifying compounds affecting biofuel quality. ELEM-SPOT addresses this by efficiently screening for oxygen and nitrogen compounds, crucial for refining processes.


ELEM-SPOT comprises a gas chromatograph mass spectrometer (GC-MS) and a combustion catalyst reactor. This combination allows for the online oxidation and decomposition of target compounds, followed by selective detection of carbon, hydrogen, nitrogen, and oxygen. With over 80% reduction in compound identification time compared to traditional methods, ELEM-SPOT streamlines biofuel analysis, minimising downtime and maximising productivity.


ELEM-SPOT offers several key features that distinguish it as a pioneering solution in biofuel analysis:


Selective Detection: ELEM-SPOT stands out as the first system capable of selectively detecting oxygen- and nitrogen-containing compounds. This capability is essential for assessing biofuel quality and preserving catalyst integrity during the refining process.


Cutting-edge R&D Support: With unparalleled sensitivity, ELEM-SPOT facilitates the analysis of trace compounds without the need for specific standards. This feature is particularly valuable for researchers engaged in the development of advanced biofuels, enabling more precise analysis of complex raw materials.


High Basic Performance: ELEM-SPOT builds upon Shimadzu’s renowned GC-MS technology, inheriting the superior performance and usability of the flagship GCMS-QP2020 NX. This ensures accurate and efficient analysis, further enhancing the reliability of biofuel assessment processes. ELEM-SPOT represents a significant advancement in biofuel analysis, supporting the transition towards sustainable energy sources. More information online: ilmt.co/PL/8xq2


62505pr@reply-direct.com


LAB ASIA - JUNE 2024


Page 1  |  Page 2  |  Page 3  |  Page 4  |  Page 5  |  Page 6  |  Page 7  |  Page 8  |  Page 9  |  Page 10  |  Page 11  |  Page 12  |  Page 13  |  Page 14  |  Page 15  |  Page 16  |  Page 17  |  Page 18  |  Page 19  |  Page 20  |  Page 21  |  Page 22  |  Page 23  |  Page 24  |  Page 25  |  Page 26  |  Page 27  |  Page 28  |  Page 29  |  Page 30  |  Page 31  |  Page 32  |  Page 33  |  Page 34  |  Page 35  |  Page 36  |  Page 37  |  Page 38  |  Page 39  |  Page 40  |  Page 41  |  Page 42  |  Page 43  |  Page 44  |  Page 45  |  Page 46  |  Page 47  |  Page 48