サーモフィッシャーサイエンティフィック株式会社／Thermo Fisher Scientific K.K. List of Products and Services

The Charged Aerosol Detector (CAD) is a highly versatile liquid chromatography detector capable of detecting all non-volatile and semi-volatile compounds. In particular, it can sensitively detect compounds that do not absorb UV light, such as lipid nanoparticles (LNPs). Additionally, the consistent responsiveness of the CAD allows for accurate assessment of the relative amounts of each lipid component. Furthermore, it is suitable for monitoring impurities and degradation products, in addition to evaluating the composition of LNPs. This application note presents examples of constructing a fast and robust UHPLC method using the Thermo Scientific Accucore C30 LC column to separate and evaluate lipid components contained in two different formulations, as well as an example using the Thermo Scientific Hypersil GOLD C8 column to effectively profile lipid components and their impurities present in raw materials. For this analysis, we used the bio-compatible Thermo Scientific Vanquish Flex and Thermo Scientific Vanquish Horizon UHPLC systems.

The HPLC-CAD system analyzes choline and its counterions, as well as impurities of choline, with high precision without the need for derivatization. The analysis conditions using mixed-mode with hydrophilic interaction chromatography (HILIC) accommodate both charged particle detection and mass spectrometry, enabling the simultaneous analysis of anions and cations, which allows for rapid and reliable detection of choline content and impurities. In this application note, we successfully detected the content of choline derivatives such as choline chloride, choline bitartrate, and choline citrate, along with their impurities, with high precision. Choline chloride, which had poor responsiveness with conventional UV detectors, can also be directly detected without the use of derivatization methods. Furthermore, the mixed-mode using HILIC accurately identifies impurities while avoiding interference from counterions. This serves as a powerful tool for ensuring safety and efficacy in quality control for dietary supplements and other products.

Foods containing sugar often have a high content of sucrose, making it difficult to accurately quantify sucrose and other disaccharides (such as lactose and maltose) in actual samples due to insufficient separation. This technical document adopts a single-loop heart-cut 2D-LC method, where lactose and maltose are fractionated in the sample loop after sucrose elutes in the first dimension, allowing for the separation and analysis of lactose and maltose in the second dimension, thereby avoiding the influence of sucrose. Additionally, the charged aerosol detector (CAD) used in the detection allows for gradient analysis, providing greater flexibility in separating contaminants in a variety of food samples, enabling efficient analysis. This achieves high-sensitivity analysis that was difficult with conventional refractive index detectors (RI). 【Features】 - Accurate analysis by separating lactose and maltose in foods containing sugar - Separation from contaminants possible with the single-loop heart-cut 2D-LC method - Charged aerosol detector (CAD) allows for gradient analysis and accommodates diverse samples - Achieves high-sensitivity analysis that is difficult with conventional refractive index detectors (RI)

This technical document introduces a rapid and quantitative analytical method for the simultaneous detection of seven types of vitamins (vitamin A, vitamin D2, vitamin D3, and four types of vitamin E isomers) in infant formula using a conventional HPLC system with a diode array detector and a fluorescence detector, following offline solid-phase extraction (SPE) and two-dimensional liquid chromatography (2D-LC) analysis. By changing the traditional liquid-liquid extraction method to solid-phase extraction, the time required for sample preparation was reduced from 3 hours to 1 hour. Additionally, by simplifying the sample preparation and optimizing the separation conditions, the influence of the matrix was reduced, allowing for the detection of target substances with higher sensitivity. ■ Simplified sample preparation through two-dimensional liquid chromatography (2D-LC) analysis ■ Simultaneous separation analysis of vitamins A, D, E, and their isomers ■ Developed a method with higher recovery rates and sensitivity than liquid-liquid extraction ■ Simplified the preprocessing process and reduced analysis time by 2.5 times ■ Reduced the influence of the matrix and provided reliable data

Many amino acids do not absorb ultraviolet (UV) light, making it difficult to improve sensitivity and selectivity with conventional analytical methods, which are also complicated and lack stability. This technical document introduces a case where a custom injection program feature in the auto-sampler of the Thermo Scientific Vanquish Core HPLC system was utilized to derivatize 20 types of amino acids simultaneously using a pre-column derivatization method for simultaneous analysis. This system enables highly accurate quantitative analysis as it is not affected by the degradation after the derivatization reaction. Validation using commercially available beverages showed that selective quantification was possible even in complex sample matrices such as food. 【Features】 ■ Simultaneous derivatization and analysis of 20 types of amino acids ■ High-precision quantitative analysis achieved through pre-column derivatization ■ Provides good linearity and high reproducibility ■ Selective detection even in complex sample matrices ■ Automation by the auto-sampler avoids human error and improves work efficiency

This technical document introduces a post-column analysis system for accurately quantifying organic acids in various beverages. The analysis column uses ion exclusion mode, and by adding BTB solution post-column, it separates and detects ten types of organic acids with good reproducibility. The e-workflow feature, which supports operations in routine analysis, is a significant advantage as it fully automates the process from device startup to measurement and report generation. The HPLC system features a compact design that incorporates a dual pump with two pumps in one module for analysis and post-column solution delivery. Additionally, even in the event of an error, the automatic switching of valves prevents the post-column reaction solution from backflowing into the analysis column, allowing for safe adoption in routine analysis. 【Features】 ■ Accurate quantification of ten types of organic acids with good reproducibility of less than 1% ■ Separation of organic acids using ion exclusion mode columns, allowing measurement without matrix effects ■ Detection limits for organic acids in this method range from a few to 7 μg/mL ■ Automation of the process from device startup to report generation with the e-workflow feature ■ Ensured safety with automatic valve switching in case of errors

This technical document presents a case study of the simultaneous analysis of polyphenols contained in beverages and health foods with high sensitivity, unaffected by other components. A 1.5 µm solid-core particle column was used for the analysis, separating 36 types of polyphenols under gradient conditions within 10 minutes. By using standard samples of each polyphenol, calibration curves and detection limits were confirmed, achieving good linearity with a coefficient of determination of over 0.997 and detection limits in the low ng/mL range. Additionally, in the spiking recovery tests conducted on multiple health foods and beverages, approximately 100% recovery was obtained, confirming accurate quantification without interference from food components. This system reliably separates and detects components with similar structures, such as polyphenols, providing reduced analysis time and high reproducibility. Furthermore, its compact design and ease of operation streamline routine analyses. 【Features】 ■ Simultaneous analysis of 36 polyphenol components ■ High separation using a 1.5 µm solid-core particle column ■ Detection limits in the low ng/mL range and high sensitivity ■ Good calibration curves with a coefficient of determination over 0.997 ■ Accurate quantification with approximately 100% recovery, unaffected by food components

This technical document presents a case study on the highly sensitive quantification of eight components, including monosaccharides, disaccharides, and sugar alcohols, in beverages using the Thermo Scientific ISQ EC single quadrupole mass spectrometer. There are various methods for sugar analysis using HPLC, but traditional differential refractive index detectors have challenges such as sensitivity issues and the inability to perform gradient analysis. Additionally, while fluorescence detectors can detect low concentrations, they require derivatization, making the sample preparation complicated. The ISQ EC single quadrupole mass spectrometer enables the quantification of sugars at low concentrations ranging from a few to several tens of ng/mL, thanks to its high sensitivity and selectivity. Furthermore, its user-friendly device and analysis software allow easy operation for both beginners and experts in mass spectrometry, streamlining routine analyses and enabling the rapid acquisition of reliable quantitative data. 【Features】 ■ High sensitivity: Detection limit of a few to several tens of ng/mL ■ High selectivity: Accurate quantification unaffected by interfering components ■ Improved operability: No complex sample preparation required

Quantitative analysis of gloss agents (saccharin and 2-butin-1,4-diol) in electroplating nickel plating solution using LC-MS helps maintain a consistent composition of the plating solution and greatly aids in the quality control of gloss agents. This method analyzes the components contained in the gloss agents under two different conditions based on their chemical composition. Additionally, samples can be easily measured by simply diluting them with water, and good results ranging from 92% to 108% have been obtained in addition-recovery tests. Furthermore, analytical conditions have been devised to achieve stable measurements over a long period. By using MS, detection is possible even at low concentrations in samples containing high concentrations of impurities, such as plating solutions, and detection is possible with both UV and MS. Measurement of not only gloss agents but also decomposition and by-products is possible, leading to further evaluation of the plating solution. For more details, please download the materials. 【Features】 ■ Quality control of gloss agents achieved through quantitative analysis using LC-MS ■ Measurement of gloss agents and decomposition/by-products is possible ■ Analysis of gloss agent components under two different conditions ■ Samples can be easily measured by simply diluting with water ■ Analytical conditions enable stable measurements over a long period

The Thermo Scientific Vanquish Duo UHPLC system is a dual LC system equipped with two independent flow paths, allowing for parallel condition testing of different separation chemistries, such as the combination of ion exchange chromatography (IEX) and ion-pair reverse phase chromatography (IP-RP), within the same system. This technical document utilizes a method scouting kit to examine various ion-pairing agents, gradient conditions, pH, and column temperatures, achieving good peak shapes for mRNA and high-sensitivity detection of impurities. By simultaneously investigating IEX and IP-RP, it is possible to understand the separation characteristics of the sample, enabling efficient determination of separation conditions. With high separation capability and sensitivity, impurities can be effectively detected during post-transcription purification, allowing for reliable mRNA purification. **Features** - Simultaneous examination of two different separation chemistries (IEX and IP-RP) using the same device. - High-sensitivity detection of impurities in post-transcription purification through mRNA profiling. - Efficient determination of suitable separation conditions using the method scouting kit. - Examination of up to 10 solvents per column type to determine appropriate mobile phase conditions.

This document introduces a method for the high-precision separation and quantification of four types of tocopherols contained in vegetable oils using the single-loop heart-cut 2D-LC method. Under the separation conditions of a reverse-phase system using a general ODS column, the separation of β-tocopherol and γ-tocopherol is difficult; however, the 2D-LC method allows for easy separation. In the first dimension, tocopherols and interfering components in the sample are separated, and in the second dimension, the four types of tocopherols (α, β, γ, δ) are separated. The repeatability of the standard samples achieved a high reproducibility with RSD < 0.1%. Additionally, measurements conducted on multiple vegetable oils showed that quantification was possible without the influence of interfering peaks. The single-loop heart-cut 2D-LC method is the simplest technique among 2D-LC methods, making it applicable for routine analysis and a useful tool for food analysis involving samples with a high content of matrix components. 【Features】 ■ High-precision separation achieved with the single-loop heart-cut 2D-LC method ■ Separation of β-tocopherol and γ-tocopherol, which is difficult in reverse-phase systems ■ Effective separation of interfering substances in food ■ High reproducibility with repeatability of RSD < 0.1%

This application introduces a method for the highly sensitive and robust analysis of L-aspartic acid and glycine impurities using the HPLC-UV-CAD method. The conventional derivatization methods used in amino acid analysis have faced challenges such as human error and variability in derivatization efficiency; however, this method provides a more reliable approach by detecting without derivatization. Additionally, it allows for the simultaneous analysis of amino acids and their impurities (especially organic acids) in a single chromatography run, significantly reducing analysis time and contributing to cost savings. Furthermore, the selectivity and limit of quantification (LOQ) of the new analytical method are equivalent to established official methods, making it useful for amino acid quality control. 【Features】 ■ Fewer errors compared to derivatization methods, offering greater robustness and reliability ■ Simultaneous detection of amino acids and organic acids using a single method, reducing analysis time and costs ■ Highly sensitive detection of amino acids through the combination of CAD and UV ■ The selectivity and limit of quantification (LOQ) of the new analytical method are equivalent to official methods *For more details, please download the PDF or feel free to contact us.

This technical document introduces an analytical method for sugar alcohols based on official methods. We measured six types of commercially available foods using the Thermo Scientific Syncronis Amino LC column in HILIC mode and the Thermo Scientific HyperREZ XP CarbohydrateCa2+ LC column in ligand exchange mode. Furthermore, the Thermo Scientific Chromeleon Chromatography Data System (CDS) report designer automatically calculates the sugar alcohol content in the food by inputting the sample volume, fixed volume, and dilution ratio during measurement. This helps prevent human errors and allows for the rapid and accurate creation of the final report. 【Features】 ■ High-precision measurement of sugar alcohols based on official methods ■ Use of specific columns in HILIC mode and ligand exchange mode ■ Automatic calculation and accurate result verification by the Chromeleon CDS report designer ■ High-reliability custom report function to prevent transcription errors and rounding errors *For more details, please download the PDF or feel free to contact us.

The development and quality control of LNP formulations require rapid and accurate identification of lipid components and evaluation of molar ratios. This technical document introduces a method for efficiently quantifying the concentration and molar ratio of each lipid in LNP formulations. The Charged Aerosol Detector (CAD) is a versatile detector for liquid chromatography that can detect all non-volatile and semi-volatile compounds. It is particularly sensitive for detecting compounds that do not absorb UV light, such as LNPs. Furthermore, by leveraging the CAD's high sensitivity, consistent responsiveness, and wide dynamic range, we aim to simplify sample preparation and facilitate method development. Additionally, by using the Thermo Scientific Accucore C30 LC column, we can effectively separate lipid components and other excipients in LNP formulations in a short time. This has also confirmed that the measurement results align well with the theoretical molar ratio%. 【Features】 ■ Rapid and accurate identification of lipid components and evaluation of molar ratios ■ Compliance support with Thermo Scientific Chromeleon CDS software ■ Effective separation with Accucore C30 LC column ■ Measurement results that align well with theoretical molar ratio%

We introduce a rapid and cost-effective method for measuring the titer of adeno-associated virus (AAV) products after downstream processing, combining size exclusion chromatography (SEC) with a highly sensitive fluorescence detector. By using the Thermo Scientific MAbPac SEC-1 column, efficient separation of AAV capsids and low molecular weight impurities is achieved, allowing for detailed monitoring of product quality attributes (PQA). This method enables rapid determination of AAV product titer after a single step of affinity purification, contributing to the evaluation of product characteristics. 【Features】 ■ Rapid and cost-effective titer measurement method ■ Measurement possible with small sample sizes due to high-sensitivity fluorescence detection ■ Efficient separation using the MAbPac SEC-1 column ■ Detailed monitoring of PQA is possible ■ Improved reproducibility with low-adsorption glass vials (RSD less than 5%) *For more details, please download the PDF or feel free to contact us.

This technical document explains an HPLC method suitable for the fingerprinting of Polysorbate 80 and proposes a simple approach to detect variations between different suppliers, grades, and manufacturing lots, as well as to identify quality issues. The Thermo Scientific ISQ EM single quadrupole mass spectrometer, with a mass range up to m/z 2,000, allows for the easy identification of a wide range of key components of polysorbates from low to high molecular weights. Additionally, the consistent response of the CAD and the reverse gradient method enable semi-quantification without standard substances, allowing for a substantial understanding of the mass balance between components with varying degrees of esterification. Please take a look at the optimal HPLC-CAD-MS method for fingerprinting Polysorbate 80. *For more details, please download the PDF or feel free to contact us.*

This content explains the benefits of combining a Charged Aerosol Detector (CAD) and a Single Quadrupole Mass Spectrometer for impurity analysis of topiramate. The HPLC-UV system is a common analytical method used in many laboratories. By combining CAD and a mass spectrometer with this system, comprehensive impurity detection becomes possible. CAD detects non-volatile and semi-volatile compounds with high sensitivity. The mass spectrometer not only confirms molecular weight but also eliminates the possibility of co-elution by verifying peak purity. Additionally, the in-source Collision-Induced Dissociation (CID) function allows for the confirmation of known impurities and the structural estimation of unknown impurities. 【Features】 ■ Identification of known impurities and molecular weight prediction and structural estimation of unknown impurities ■ High-precision impurity profiling through combination with a Single Quadrupole Mass Spectrometer ■ Peak identification based on m/z and elimination of co-elution possibilities ■ CAD has high sensitivity and a wide dynamic range, accommodating gradient analysis *For more details, please download the PDF or feel free to contact us.

This technical document introduces a method for efficiently analyzing ten types of ultraviolet filter components contained in five types of sunscreen using a UHPLC system under a single analytical condition. The results of the validation of the method's accuracy and precision showed that the peak area RSD was below 1.10%, and the retention time RSD was below 0.50%, indicating high precision. Additionally, by transitioning the existing analytical method to the UHPLC method, we reduced solvent consumption and waste generation, and replaced tetrahydrofuran (THF) used in the sample solution with a more environmentally friendly solvent, thereby reducing the environmental impact. 【Features】 ■ Efficient analysis of ten types of ultraviolet filter components under a single analytical condition ■ High precision with peak area RSD below 1.10% and retention time RSD below 0.50% ■ Reduction of solvent consumption and waste generation through the UHPLC method ■ Thorough evaluation of accuracy, linearity, and precision ■ Use of environmentally friendly solvents to reduce environmental impact *For more details, please download the PDF or feel free to contact us.

This document presents guidelines for smoothly integrating the Thermo Scientific Vanquish HPLC/UHPLC system with Waters Corporation's Empower 3 Chromatography Data System (CDS), along with case studies from Contract Development and Manufacturing Organizations (CDMOs). By introducing a new LC system, we were able to significantly reduce system suitability failures (SSF), extend column life, and improve operational efficiency in a cGMP environment. Additionally, we will provide specific examples of the method transfer process between analytical instruments from different manufacturers. 【Features】 ■ Significant reduction in system suitability failures due to equipment upgrades ■ Intelligent features that greatly simplify the method transfer process ■ Achieving reproducible and robust separations with very high stability, precision, and accuracy ■ Thermo Scientific Vanquish Duo UHPLC system: significantly improved throughput and productivity with two independent flow paths

The Thermo Scientific Vanquish Split Sampler significantly reduces the effort and time associated with traditional manual dilution by allowing users to set up an automated dilution program for samples. This technical note verifies the accuracy and reproducibility of automated dilution of standard solutions through the measurement of DNPH-derivatized formaldehyde. Custom injections using user-defined programs enable easy execution of multi-step dilutions and automation of calibration curve creation. Furthermore, by utilizing the e-workflow of Thermo Scientific Chromeleon CDS, the entire process from analysis to report generation can be greatly streamlined, enhancing the reliability of analyses and increasing laboratory productivity. **Features** - High reproducibility and accuracy through automated dilution - Flexible dilution settings with custom injection programs - Automation of analysis with Chromeleon CDS e-workflow, eliminating human error - Significant reduction in effort and time compared to manual dilution - High-precision preparation capable of up to 20-fold dilution

In the quality control of plating solutions, the quantification of additives is an important factor. In particular, the quantification of sulfur-containing compounds that contribute to film quality adjustment and prevention of liquid decomposition is essential. This document provides accurate and highly reproducible analytical results using a method that does not require complex pretreatment. For the HPLC measurement conditions, an ion-pair reagent was added to the mobile phase, and an injection method was adopted to reduce the influence of sample impurities. As a result, sharp peaks can be obtained without being affected by metals and inorganic ions. Additionally, by comparing the chromatograms of the standard solution and the plating solution sample, it can be confirmed that the concentration of additives is maintained consistently. Furthermore, as an evaluation of reproducibility, the same sample was measured five times, demonstrating high reproducibility. 【Features】 ■ High-sensitivity quantitative analysis without complex pretreatment ■ High reproducibility using ion-pair reagents ■ Injection method that reduces the influence of metals and inorganic ions ■ Accurate quantification through comparison of standard solutions and actual samples ■ Highly reproducible analytical results

This application note introduces the analysis of polymer compounds using size exclusion chromatography (SEC) with the Vanquish Charged-Aerosol Detector (CAD). The CAD has approximately 100 times the sensitivity compared to a differential refractive index detector (RI) and is a versatile detector capable of accurately detecting semi-volatile and non-volatile components. Its wide dynamic range and excellent stability make it ideal for analyzing molecular weight distributions. This document discusses the detection of calibration standard samples and calibration curves, comparisons of sensitivity, and changes in retention time due to sample concentration, using polystyrene samples to compare RI and explain the characteristics and advantages of CAD. 【Features】 ■ High sensitivity detection: CAD achieves approximately 100 times the sensitivity compared to RI ■ Wide dynamic range: Accurately measures samples of diverse molecular weights ■ Detection in the polymer range: Effectively detects polymers with molecular weights up to 400,000 ■ Low concentration measurement: Minimal concerns about carryover or column loading even at low concentrations ■ Stability: Provides rapid stabilization and consistent results

The HPLC Charged Aerosol Detector (CAD) system is a solution that allows for the rapid and highly accurate quantification of lipids in liposome formulations. This technical sample presents the results of validation using the ASTM method across 12 system configurations at 4 sites (Thermo Scientific Vanquish Flex UHPLC and Vanquish Core HPLC). Reliable results were obtained across all systems, and they passed the system suitability tests. Additionally, excellent separation performance was confirmed using the Thermo Scientific Hypersil GOLD column. 【Features】 ■ High precision quantification: Quantification is possible in the range of 0.2 to 300 µg/g using linear, quadratic, and log-log calibration curves. ■ High reliability: Passed system suitability tests at all sites. ■ Excellent performance: High separation capability and rapid analysis with the Hypersil GOLD column. ■ Efficient analysis: Consistent responsiveness and high-precision results obtained quickly.

The Thermo Scientific Vanquish Fraction Collector (VFC) is a system that enables peak-triggered fractionation using UV, CAD, and MS detectors. With CAD and MS detection, it has become possible to fractionate and evaluate unknown components and impurities that were difficult to detect with conventional UV detection. This application note explains the features of the VFC equipped with innovative valve technology, using examples from plant extracts. It provides detailed practical know-how from parameter settings for accurate fractionation to applications on a semi-preparative scale. 【Features】 ■ Low contamination: Cleaning functions for needle and buffer loop ■ High precision fractionation: Automatic calculation of delay volume, peak-triggered fractionation ■ Excellent versatility: Diverse fractionation settings using UV, CAD, and MS detection ■ High reliability: Fractionation enabled by innovative valve technology