In this interview, we asked Hemanth Kumar Chanduluru from the SRM Institute of Science and Technology in Kattankulathur, India, multiple questions regarding sustainability in analytical separation methods. Within the context of developing chromatographic analytical methods, the concept of sustainability refers to designing methods that minimize the environmental impact while maintaining high efficiency and effectiveness in separation and analysis processes. This involves several considerations including reducing usage volume of solvents, energy efficiency, waste reduction, Green chemistry principles, and overall life cycle assessment for sustainability of the entire analytical measurement and equipment process. Along these sustainability efforts there are useful processes and metrics that may be used to objectively evaluate progress, including Analytical Quality by Design (AQbD), and up-to-date ChlorTox Scale, greenness, and whiteness score toolsets.
Could you elaborate on how Integrative AQbD (Analytical Quality by Design) principles were applied in the development of the RP-HPLC method for simultaneous separation of triple antihypertensive combination therapy (1)?
Integrative Analytical Quality by Design (AQbD) principles offer a systematic approach to method development, focusing on understanding the relationships between critical method parameters (CMPs) and critical method attributes (CMAs) to ensure robust and reliable analytical methods. When applied to the development of a reversed-phase high performance liquid chromatography (RP-HPLC) method for simultaneous separation of a triple antihypertensive combination therapy, several steps can be followed:
1. Defining the Quality Target Product Profile (QTPP): This involves specifying the desired characteristics of the analytical method, such as specificity, accuracy, precision, linearity, range, and robustness.
2. Identification of Critical Method Attributes (CMAs): These are the characteristics of the method that have a significant impact on its performance. For an RP-HPLC method, CMAs may include resolution, retention time, and peak shape for each component of the triple antihypertensive combination.
3. Selection of Critical Method Parameters (CMPs): These are the parameters of the analytical method that can influence CMAs. Examples of CMPs for RP-HPLC may include mobile phase composition, column type and dimensions, temperature, flow rate, and gradient program.
4. Design of Experiments (DoE): Using statistical design principles, experiments are conducted to systematically evaluate the effects of different levels of CMPs on CMAs. This helps in understanding the design space of the method and identifying the optimal conditions.
5. Risk Assessment and Mitigation: Potential risks to method performance are identified and addressed through risk assessment tools such as Failure Mode and Effects Analysis (FMEA). Strategies are developed to mitigate these risks, ensuring method robustness.
6. Method Development and Optimization: Based on the results of DoE and risk assessment, the method is developed and optimized to achieve the desired CMAs. This may involve adjusting CMPs within the established design space to maximize method performance.
7. Method Validation: Once the method is developed, it undergoes validation to demonstrate its suitability for its intended purpose. This includes assessing parameters such as specificity, accuracy, precision, linearity, range, and robustness, in accordance with regulatory guidelines.
8. Continuous Improvement and Lifecycle Management: After validation, the method is subject to continuous monitoring and improvement throughout its lifecycle. Any changes to method conditions or procedures are systematically evaluated to ensure that method performance remains within acceptable limits.
By following these steps, integrative AQbD principles can be effectively applied in the development of an RP-HPLC method for simultaneous separation of triple antihypertensive combination therapy, ensuring that the method is robust, reliable, and fit for its intended purpose.
What specific considerations were taken into account to ensure the sustainability of the RP-HPLC method during its development and validation process?
To ensure the sustainability of the RP-HPLC method during its development and validation process, several specific considerations can be taken into account:
1. Green Chemistry Principles: Integration of green chemistry principles involves minimizing the use of hazardous materials, reducing waste generation, and optimizing resource utilization. This can be achieved by selecting eco-friendly solvents, reducing solvent consumption, and employing efficient chromatographic conditions that minimize environmental impact.
2. Economic Viability: Considering the economic aspects of method development and validation is crucial for sustainability. This includes evaluating the cost-effectiveness of reagents, consumables, and equipment, as well as assessing the scalability of the method to ensure its feasibility for routine use in the laboratory.
3. Energy Efficiency: Optimizing method conditions to minimize energy consumption contributes to sustainability. This may involve reducing run times, optimizing column temperature, and selecting appropriate instrument settings to conserve energy while maintaining method performance.
4. Resource Conservation: Minimizing resource consumption, such as solvent usage and sample volume, helps conserve valuable resources and reduces waste generation. Techniques such as microextraction and miniaturization can be employed to reduce sample and solvent volumes without compromising method sensitivity or reliability.
5. Robustness and Reliability: Developing a robust and reliable method ensures its long-term sustainability by reducing the need for frequent method optimization and troubleshooting. Implementing quality control measures and conducting thorough method validation help establish the method's reliability and ensure consistent performance over time.
6. Compliance with Regulatory Guidelines: Ensuring compliance with regulatory guidelines and standards is essential for the sustainability of the method, as it facilitates acceptance and adoption by regulatory authorities. Adhering to Good Laboratory Practices (GLP) and relevant regulatory requirements during method development and validation is critical for achieving regulatory approval and maintaining method sustainability.
7. Continuous Improvement and Optimization: Implementing a process of continuous improvement and optimization throughout the method lifecycle enhances its sustainability. This involves monitoring method performance, identifying areas for improvement, and implementing changes to optimize method efficiency, reliability, and environmental impact.
By incorporating these considerations into the development and validation process of the RP-HPLC method, sustainability can be effectively addressed, ensuring that the method remains environmentally friendly, economically viable, and operationally efficient throughout its lifecycle.
How does the use of a special C18 (2) column contribute to the sustainability aspect of the developed method compared to other column options?
The choice of the C18 column can contribute to the sustainability aspect of the developed RP-HPLC method compared to other column options in several ways:
1. Longevity and Durability: Our selected C18 columns are known for their robustness and longevity. These columns typically exhibit excellent column stability and can withstand a wide range of mobile phase conditions and sample matrices without significant degradation. As a result, they have a longer operational lifetime, reducing the frequency of column replacement and minimizing waste generation.
2. Reduced Solvent Consumption: Our C18 columnwas designed to provide efficient chromatographic separations at lower solvent volumes. Their high efficiency and resolving power allow for shorter analysis times and reduced solvent consumption per analysis. This not only saves solvent costs but also contributes to environmental sustainability by minimizing solvent usage and waste generation.
3. Lower Energy Consumption: Our C18 column used in RP-HPLC typically requires lower column temperatures for optimal performance compared to other column options. This can lead to reduced energy consumption during chromatographic runs, contributing to overall energy efficiency and sustainability.
4. Compatibility with Green Solvents: Our selected columns are often compatible with eco-friendly solvents, such as aqueous-organic mobile phases containing lower concentrations of organic solvents or alternative green solvents. By enabling the use of greener solvent systems, these columns help reduce the environmental impact associated with solvent usage in analytical laboratories.
5. High Analytical Performance: Our C18 columns are known for their high analytical performance, including excellent peak shapes, resolution, and reproducibility. Their superior chromatographic properties minimize the need for method optimization and troubleshooting, leading to more efficient and sustainable analytical workflows.
6. Versatility and Application Range: Our selected C18 columns are versatile and suitable for a wide range of applications, including pharmaceutical analysis, environmental monitoring, food safety testing, and more. Their broad applicability reduces the need for multiple column types, simplifying laboratory operations and inventory management, which in turn enhances sustainability by minimizing resource consumption and waste generation.
Overall, the use of the appropriate C18 columns in the developed RP-HPLC method contributes to sustainability by improving column longevity, reducing solvent and energy consumption, enabling the use of green solvents, enhancing analytical performance, and simplifying laboratory operations. These factors collectively support environmentally friendly and economically viable analytical practices, aligning with sustainability goals in analytical chemistry.
Can you discuss the rationale behind selecting ethanol (EtOH) and KH2PO4 as components of the mobile phase, particularly in terms of their environmental impact and sustainability?
Ethanol can be derived from renewable resources such as biomass (for example, corn, sugarcane, or cellulosic materials). Unlike fossil-based solvents, which are finite resources, ethanol production can be sustainable when sourced from renewable feedstocks.
Ethanol is also biodegradable, meaning it can be broken down into harmless byproducts by microorganisms in the environment. This reduces its environmental impact compared to non-biodegradable solvents.
Third, ethanol is generally less toxic than some other organic solvents commonly used in chromatography, such as acetonitrile or methanol. Lower toxicity levels contribute to reduced environmental and health risks associated with solvent handling and disposal.
And finally, ethanol typically has lower volatile organic compound (VOC) emissions compared to certain other organic solvents. This is beneficial for air quality and reduces the environmental footprint associated with solvent evaporation during chromatographic analyses.
For KH2PO4, it serves as a buffer salt in the mobile phase, helping to control and maintain the pH of the solution. Phosphorus is an essential nutrient for plant growth, and its use in the form of KH2PO4 can contribute to the fertilization of soils when disposed of responsibly.
Second, KH2PO4 is biodegradable and does not persist in the environment, unlike some other buffer salts or additives commonly used in chromatography. Its biodegradability ensures minimal long-term environmental impact.
Third, potassium dihydrogen phosphate is generally considered to have low environmental toxicity. When disposed of properly, it poses minimal risks to aquatic organisms and ecosystems compared to certain other buffer salts or additives.
And finally, the production and use of KH2PO4 typically have a relatively low environmental footprint compared to some other buffer salts or additives. This is important for sustainability, as it reduces resource consumption and energy usage associated with manufacturing and transportation.
What strategies were employed to optimize the separation time while maintaining the environmental sustainability of the method?
To optimize the separation time while maintaining the environmental sustainability of the RP-HPLC method, several strategies can be employed:
1. Column Selection and Dimensions: Choosing an appropriate column type and dimensions can significantly impact separation time. Columns with smaller particle sizes and shorter lengths typically offer higher efficiency and faster separations. By selecting a column optimized for rapid separations, overall analysis time can be reduced, leading to lower solvent consumption and energy usage.
2. Mobile Phase Composition and Gradient Program: Optimization of the mobile phase composition and gradient program plays a crucial role in achieving fast and efficient separations. By carefully selecting solvent systems and gradient profiles, it is possible to improve peak resolution and shorten analysis time without compromising separation quality. Additionally, using eco-friendly solvents with lower environmental impact, such as ethanol, and optimizing gradient conditions to minimize solvent usage contribute to environmental sustainability.
3. Optimized Flow Rate: Adjusting the flow rate of the mobile phase can influence separation time without compromising chromatographic performance. Higher flow rates generally result in faster elution times but may compromise resolution. By optimizing the flow rate within the range that maintains adequate resolution while minimizing analysis time, overall solvent consumption and energy usage can be reduced, enhancing environmental sustainability.
4. Temperature Control: Proper temperature control of the chromatographic system can affect separation efficiency and analysis time. Maintaining a stable column temperature within an optimal range can improve chromatographic performance and reduce the time required for equilibration between runs. Additionally, optimizing column temperature can help achieve faster elution times without sacrificing resolution, contributing to shorter analysis times and reduced energy consumption.
5. Method Development using Quality by Design (QbD) Principles: Applying Quality by Design (QbD) principles during method development allows for systematic optimization of critical method parameters while considering environmental sustainability. By utilizing experimental design techniques, such as Design of Experiments (DoE), and conducting risk assessments, it is possible to identify the most influential factors affecting separation time and develop strategies to optimize them while minimizing environmental impact.
6. Continuous Monitoring and Improvement: Implementing a process of continuous monitoring and improvement allows for ongoing optimization of the method to further reduce separation time and enhance environmental sustainability. By regularly evaluating method performance and identifying opportunities for refinement, it is possible to achieve incremental improvements in efficiency and environmental impact over time.
By employing these strategies, it is possible to optimize the separation time of the RP-HPLC method while maintaining environmental sustainability, thereby reducing solvent consumption, energy usage, and overall environmental impact associated with chromatographic analysis.
Could you explain how the gradient elution system was designed to enhance both separation efficiency and greenness of the method?
Designing a gradient elution system in an RP-HPLC method to enhance both separation efficiency and greenness involves optimizing the mobile phase composition and gradient profile to achieve efficient analyte separation while minimizing environmental impact. Here's how this can be achieved:
1. Mobile Phase Composition:
2. Gradient Profile Optimization:
3. Monitoring and Control:
By incorporating these design principles, a gradient elution system can be tailored to enhance both separation efficiency and greenness in an RP-HPLC method. Optimization of mobile phase composition, gradient profile, and control parameters enables rapid, efficient separations with reduced environmental impact, aligning with sustainability goals in analytical chemistry.
How does the use of a specific temperature (35 °C) during chromatographic analysis align with the concept of sustainability in the developed method?
The use of a specific temperature, such as 35 °C, during chromatographic analysis can align with the concept of sustainability in the developed method in several ways:
1. Energy Efficiency: Maintaining a constant temperature during chromatographic analysis helps optimize energy efficiency. By operating the chromatographic system at a moderate temperature, such as 35 °C, excessive energy consumption associated with extreme temperature settings can be avoided. This contributes to overall energy conservation and reduces the environmental footprint of the analytical method.
2. Column Stability and Longevity: Operating the chromatographic column at a controlled temperature helps maintain column stability and prolong its operational lifetime. Fluctuations in temperature can lead to column degradation and reduced chromatographic performance over time. By setting a specific temperature, such as 35 °C, within the optimal range for the column material, stability is enhanced, minimizing the frequency of column replacement and reducing waste generation.
3. Reproducibility and Robustness: Consistent temperature control ensures reproducible chromatographic results and method robustness. Temperature variations can affect analyte retention times, peak shapes, and resolution, leading to inconsistencies in analysis. By maintaining a constant temperature, method variability is minimized, enhancing the reliability and reproducibility of the analytical method, which is crucial for sustainable analytical practices.
4. Reduced Solvent Consumption: Stable chromatographic conditions, including temperature, contribute to efficient solvent usage. Fluctuations in temperature can impact solvent evaporation rates and elution times, affecting overall solvent consumption during chromatographic analysis. By optimizing temperature control, solvent usage can be minimized, leading to reduced waste generation and environmental impact associated with solvent disposal.
5. Method Transferability and Standardization: Setting a specific temperature for chromatographic analysis promotes method transferability and standardization across different laboratories and instruments. Consistent temperature conditions facilitate reproducibility of results and ensure comparability of data generated from different systems. This promotes efficient knowledge sharing and collaboration within the scientific community, contributing to sustainable practices in analytical chemistry.
Overall, the use of a specific temperature, such as 35 °C, during chromatographic analysis aligns with sustainability principles by optimizing energy efficiency, enhancing column stability, improving method reproducibility, reducing solvent consumption, and promoting method transferability. These considerations collectively support environmentally friendly and economically viable analytical practices, contributing to sustainable development in analytical chemistry.
Can you describe the role of the Analytical Method Greenness Score (AMGS) in evaluating the environmental impact of the developed RP-HPLC method?
The Analytical Method Greenness Score (AMGS) serves as a quantitative tool for assessing the environmental impact of an analytical method, such as the developed RP-HPLC method. It provides a systematic approach to evaluate various aspects of method design, operation, and waste generation, enabling researchers to make informed decisions to minimize environmental impact. Here's how the AMGS can be used to evaluate the environmental sustainability of the developed RP-HPLC method:
1. Component Assessment: The AMGS considers the environmental impact of individual components used in the analytical method, including solvents, reagents, buffers, and consumables. It evaluates factors such as toxicity, biodegradability, resource consumption, and emissions associated with each component. For the RP-HPLC method, the AMGS would assess the greenness of solvents (for example, ethanol), buffer salts (for example, KH2PO4), and other chemicals used in the mobile phase and sample preparation.
2. Energy Consumption: AMGS considers the energy consumption associated with method operation, including instrument usage, temperature control, and data processing. It evaluates the efficiency of energy usage and identifies opportunities to minimize energy consumption while maintaining method performance. For the RP-HPLC method, the AMGS would assess the energy usage during chromatographic analysis, including column heating, pump operation, and detector operation.
3. Waste Generation: The AMGS evaluates the amount and nature of waste generated during method operation, including solvent waste, consumable waste, and chemical waste. It considers factors such as solvent usage, sample volume, consumable consumption, and waste disposal practices. For the RP-HPLC method, the AMGS would assess the volume of solvent waste generated during chromatographic analysis, as well as the disposal practices for used columns, consumables, and chemicals.
4. Life Cycle Analysis: The AMGS may incorporate life cycle analysis principles to assess the environmental impact of the method throughout its entire lifecycle, from raw material extraction and manufacturing to method operation and disposal. It considers the environmental footprint associated with each stage of the method lifecycle and identifies opportunities to reduce environmental impact at each stage. For the RP-HPLC method, the AMGS would assess the environmental impact of column manufacturing, instrument operation, sample preparation, analysis, and waste disposal.
5. Scoring and Ranking: Based on the assessment of various environmental factors, the AMGS assigns a numerical score to the analytical method, indicating its overall greenness. This score allows for comparison and ranking of different methods based on their environmental impact. Researchers can use the AMGS to identify areas for improvement and implement strategies to enhance the environmental sustainability of the method. For the RP-HPLC method, the AMGS would provide a quantitative measure of its environmental impact, guiding efforts to minimize resource consumption, energy usage, and waste generation while maintaining analytical performance.
In summary, the Analytical Method Greenness Score (AMGS) plays a crucial role in evaluating the environmental impact of the developed RP-HPLC method by assessing factors such as component greenness, energy consumption, waste generation, and life cycle analysis. It provides researchers with a quantitative tool to assess and optimize the environmental sustainability of analytical methods, contributing to the development of greener analytical practices.
In what ways does the RP-HPLC method address concerns regarding solvent selection and waste generation, considering the principles of green analytical chemistry?
The RP-HPLC method can address concerns regarding solvent selection and waste generation in alignment with the principles of green analytical chemistry in several ways:
1. Solvent Selection:
2. Waste Generation Reduction:
3. Green Analytical Method Development:
4. Regulatory Compliance:
Overall, the RP-HPLC method addresses concerns regarding solvent selection and waste generation by the principles of green analytical chemistry by prioritizing the use of eco-friendly solvents, minimizing solvent consumption, optimizing method efficiency, and ensuring regulatory compliance. These efforts collectively contribute to the development of greener analytical practices that promote environmental sustainability.
Could you discuss how the concept of "whiteness" was integrated into the evaluation of the sustainability profile of the developed method?
The concept of "whiteness" in the evaluation of the sustainability profile of the developed RP-HPLC method likely refers to the consideration of aspects related to the environmental impact of materials used in the method, particularly in terms of their purity, cleanliness, and ecological footprint. Integrating the concept of whiteness into the evaluation of sustainability involves assessing the environmental impact of materials and processes involved in method development and operation, with a focus on promoting purity, cleanliness, and minimal ecological disturbance. Here's how this concept might be integrated into the evaluation of the sustainability profile of the developed method:
1. Purity of Materials:
2. Cleanliness of Processes:
3. Ecological Footprint Reduction:
Overall, integrating the concept of whiteness into the evaluation of the sustainability profile of the developed RP-HPLC method involves assessing the purity, cleanliness, and ecological responsibility of materials and processes involved in method development and operation. By prioritizing purity, cleanliness, and minimal ecological disturbance, the method aims to promote environmental sustainability and contribute to a cleaner and more sustainable laboratory environment.
How does the application of the Chloroform-oriented Toxicity Estimation Scale (ChlorTox Scale) indicator contribute to assessing the environmental sustainability of the chemicals used in the method?
The ChlorTox Scale (3) is an indicator used to assess the environmental sustainability of chemicals based on their potential to generate chlorinated organic compounds and contribute to environmental pollution. It evaluates the chlorine content of chemicals and assigns a score based on their potential to form chlorinated by-products during use and disposal. The application of the ChlorTox Scale in assessing the environmental sustainability of the chemicals used in the developed method can provide valuable insights into their environmental impact and help identify opportunities for improvement. Here's how the ChlorTox Scale contributes to assessing the environmental sustainability of chemicals used in the method:
1. Quantification of Environmental Impact:
2. Comparison and Ranking:
3. Guidance for Chemical Selection:
4. Promotion of Green Chemistry Principles:
Overall, the application of the ChlorTox Scale in assessing the environmental sustainability of chemicals used in the developed method provides a systematic approach to evaluating their potential to generate chlorinated organic compounds and contribute to environmental pollution. By considering ChlorTox scores in chemical selection and method optimization, researchers can minimize the environmental impact of analytical methods and promote sustainable laboratory practices.
Can you elaborate on the significance of the Greenness Index tool through spider plots (radar charts) in evaluating the environmental sustainability of solvents and chemicals employed in the method?
The Greenness Index tool, often represented through spider plots, is a valuable tool for evaluating the environmental sustainability of solvents and chemicals employed in analytical methods, including the developed RP-HPLC method. Spider plots provide a visual representation of various environmental parameters, allowing for a comprehensive assessment of the greenness or environmental impact of different chemicals.
Here's how the Greenness Index tool through spider plots can be significant in evaluating the environmental sustainability of solvents and chemicals used in the method:
1. Multi-Parameter Evaluation:
2. Comparative Analysis:
3. Identification of Strengths and Weaknesses:
4. Optimization of Method Components:
5. Communication of Sustainability Performance:
Overall, the Greenness Index tool through spider plots is significant in evaluating the environmental sustainability of solvents and chemicals employed in analytical methods by providing a multi-parameter evaluation, enabling comparative analysis, identifying strengths and weaknesses, optimizing method components, and facilitating communication of sustainability performance. Incorporating spider plots into sustainability assessments enhances the transparency, comprehensiveness, and effectiveness of efforts to promote green chemistry and sustainable laboratory practices.
(1) Kannaiah, K. P.; Chanduluru, H. K.; Lotfy, H. M.; et al. Integrative AQbD, Up-to-date Greenness, and Whiteness Tools for Evaluation of a Sustainable RP-HPLC Method Used for Simultaneous Separation of Triple Antihypertensive Combination Therapy as a Model. Sustain. Chem. Pharm. 2023, 36, 101288. DOI: 10.1016/j.scp.2023.101288
(2) An Eclipse Plus C18 column was selected for this research: Home Page. https://www.agilent.com/cs/library/specifications/public/820114-002.pdf (accessed 2024-03-27).
(3) Nowak, P. M.; Wietecha-Posłuszny, R.; Płotka-Wasylka, J.; Tobiszewski, M. How to Evaluate Methods Used in Chemical Laboratories in Terms of the Total Chemical Risk?–a ChlorTox Scale. Green Analytical Chemistry 2023, 5, 100056. DOI: 10.1016/j.greeac.2023.100056
The Chromatographic Society 2025 Martin and Jubilee Award Winners
December 6th 2024The Chromatographic Society (ChromSoc) has announced the winners of the Martin Medal and the Silver Jubilee Medal for 2025. Professor Bogusław Buszewski of Nicolaus Copernicus University in Torun, Poland, has been awarded the prestigious Martin Medal, and the 2025 Silver Jubilee Medal has been awarded to Elia Psillakis of the Technical University of Crete in Greece.
Inside the Laboratory: Using GC–MS to Analyze Bio-Oil Compositions in the Goldfarb Group
December 5th 2024In this edition of “Inside the Laboratory,” Jillian Goldfarb of Cornell University discusses her laboratory’s work with using gas chromatography–mass spectrometry (GC–MS) to characterize compounds present in biofuels.