Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin

Abstract The International Agency for Research on Cancer categorized nitrosamines as potential or probable human carcinogens. Hence, nitrosamine impurities should be controlled to minimize the cancer risk. Sustainable Development Goal 9 (SDG9: Industry, Innovation, and Infrastructure), Target 9.4 wa...

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Main Authors: Sriram Pepakayala, Venkata Nadh Ratnakaram, Yuvaraj Zunjarrao, Yogesh Kumar Lohia
Format: Article
Language:English
Published: BMC 2025-05-01
Series:BMC Chemistry
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Online Access:https://doi.org/10.1186/s13065-025-01503-4
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author Sriram Pepakayala
Venkata Nadh Ratnakaram
Yuvaraj Zunjarrao
Yogesh Kumar Lohia
author_facet Sriram Pepakayala
Venkata Nadh Ratnakaram
Yuvaraj Zunjarrao
Yogesh Kumar Lohia
author_sort Sriram Pepakayala
collection DOAJ
description Abstract The International Agency for Research on Cancer categorized nitrosamines as potential or probable human carcinogens. Hence, nitrosamine impurities should be controlled to minimize the cancer risk. Sustainable Development Goal 9 (SDG9: Industry, Innovation, and Infrastructure), Target 9.4 was realized by developing a simple and straightforward LC-APCI-MS/MS method for simultaneous determination of potential nitrosamines: N-nitroso dimethylamine, N-nitroso di isopropyl amine, N-nitroso isopropyl ethylamine and N-nitroso methyl aminopyridine (NDMA, NDIPA, NIPEA and NMAP) in Sitagliptin Phosphate Monohydrate API (STG-API), an antidiabetic medication for type-2 diabetes and validated as per ICH (Q2 R2) guidelines. As a part of SDG9 (Industry, Innovation, and Infrastructure), Target 9.4, greater adoption of clean and environmentally sound technologies is possible by using the current analytical method in view of (a) less consumption of solvents as it has a lower run time of 18 min (b) at-line sample collection (c) simple sample preparation (d) sample preparation process that does not require derivatization or sample extraction. The optimized LC method conditions are an Agilent Poroshell EC C18 column, a flow rate of 0.6 mL/min, an injection volume of 40 µL, a column oven temperature set at 50 °C, and a sample cooler temperature set at 6 °C. A gradient method was adopted using a mixture of mobile phases (Solvent A: 0.1% formic acid in water and Solvent B: a mixture of 50% Methanol and 50% Acetonitrile). The response was obtained with Multiple Reaction Monitoring (MRM) in APCI mode. The current method can quantify NDMA, NIPEA, NDIPA, and NMAP to a lower level of 74.19 ng/g, 19.62 ng/g, 20.36 ng/g, and 13.65 ng/g, respectively, which is less than 10% of the specified limits. Good linearity in the range of Limit of Quantitation (LOQ) to 150% of the specification limit was observed, with correlation coefficients higher than 0.996. The recoveries were over the range between 90.23 and 103.36%. Assessed the method’s eco-friendliness by greenness tools like the Modified Green Analytical Procedure Index (MoGAPI), ComplexMoGAPI, Analytical GREEnness (AGREE), and Analytical Eco-Scale, and found it to be eco-friendly. Graphical Abstract
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spelling doaj-art-a6519afb2d514b09a34a91044f70d6282025-08-20T03:27:10ZengBMCBMC Chemistry2661-801X2025-05-0119112110.1186/s13065-025-01503-4Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptinSriram Pepakayala0Venkata Nadh Ratnakaram1Yuvaraj Zunjarrao2Yogesh Kumar Lohia3Analytical Research and Development, IOL Chemicals and Pharmaceuticals LtdIndustrial Chemical Product Development and Analysis Centre, Department of Chemistry, GITAM School of Science, GITAM Deemed to be University – Bengaluru CampusAnalytical Research and Development, IOL Chemicals and Pharmaceuticals LtdAnalytical Research and Development, IOL Chemicals and Pharmaceuticals LtdAbstract The International Agency for Research on Cancer categorized nitrosamines as potential or probable human carcinogens. Hence, nitrosamine impurities should be controlled to minimize the cancer risk. Sustainable Development Goal 9 (SDG9: Industry, Innovation, and Infrastructure), Target 9.4 was realized by developing a simple and straightforward LC-APCI-MS/MS method for simultaneous determination of potential nitrosamines: N-nitroso dimethylamine, N-nitroso di isopropyl amine, N-nitroso isopropyl ethylamine and N-nitroso methyl aminopyridine (NDMA, NDIPA, NIPEA and NMAP) in Sitagliptin Phosphate Monohydrate API (STG-API), an antidiabetic medication for type-2 diabetes and validated as per ICH (Q2 R2) guidelines. As a part of SDG9 (Industry, Innovation, and Infrastructure), Target 9.4, greater adoption of clean and environmentally sound technologies is possible by using the current analytical method in view of (a) less consumption of solvents as it has a lower run time of 18 min (b) at-line sample collection (c) simple sample preparation (d) sample preparation process that does not require derivatization or sample extraction. The optimized LC method conditions are an Agilent Poroshell EC C18 column, a flow rate of 0.6 mL/min, an injection volume of 40 µL, a column oven temperature set at 50 °C, and a sample cooler temperature set at 6 °C. A gradient method was adopted using a mixture of mobile phases (Solvent A: 0.1% formic acid in water and Solvent B: a mixture of 50% Methanol and 50% Acetonitrile). The response was obtained with Multiple Reaction Monitoring (MRM) in APCI mode. The current method can quantify NDMA, NIPEA, NDIPA, and NMAP to a lower level of 74.19 ng/g, 19.62 ng/g, 20.36 ng/g, and 13.65 ng/g, respectively, which is less than 10% of the specified limits. Good linearity in the range of Limit of Quantitation (LOQ) to 150% of the specification limit was observed, with correlation coefficients higher than 0.996. The recoveries were over the range between 90.23 and 103.36%. Assessed the method’s eco-friendliness by greenness tools like the Modified Green Analytical Procedure Index (MoGAPI), ComplexMoGAPI, Analytical GREEnness (AGREE), and Analytical Eco-Scale, and found it to be eco-friendly. Graphical Abstracthttps://doi.org/10.1186/s13065-025-01503-4NitrosaminesGenotoxic impuritiesLC–MS/MSMethod developmentMultiple reaction monitoringValidation
spellingShingle Sriram Pepakayala
Venkata Nadh Ratnakaram
Yuvaraj Zunjarrao
Yogesh Kumar Lohia
Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
BMC Chemistry
Nitrosamines
Genotoxic impurities
LC–MS/MS
Method development
Multiple reaction monitoring
Validation
title Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
title_full Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
title_fullStr Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
title_full_unstemmed Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
title_short Green and sustainable LC-APCI-MS/MS method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication: sitagliptin
title_sort green and sustainable lc apci ms ms method for simultaneous quantification of genotoxic nitrosamines in antidiabetic medication sitagliptin
topic Nitrosamines
Genotoxic impurities
LC–MS/MS
Method development
Multiple reaction monitoring
Validation
url https://doi.org/10.1186/s13065-025-01503-4
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