Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals
The recent detection of N-nitroso-atenolol, a mutagenic and potentially carcinogenic impurity in atenolol-based pharmaceuticals, has raised serious safety concerns and emphasized the need for stringent analytical control. This study developed and validated a highly sensitive LC-MS/MS method for quan...
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2025-05-01
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| author | Soonho Kwon Sang-Hyun Ahn Yongha Chang Joon-Sang Park Hwangeui Cho Jung-Bok Kim |
| author_facet | Soonho Kwon Sang-Hyun Ahn Yongha Chang Joon-Sang Park Hwangeui Cho Jung-Bok Kim |
| author_sort | Soonho Kwon |
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| description | The recent detection of N-nitroso-atenolol, a mutagenic and potentially carcinogenic impurity in atenolol-based pharmaceuticals, has raised serious safety concerns and emphasized the need for stringent analytical control. This study developed and validated a highly sensitive LC-MS/MS method for quantifying N-nitroso-atenolol in both active pharmaceutical ingredients (APIs) and finished products. Quantification was carried out using multiple reaction monitoring (MRM) under positive-mode electrospray ionization (ESI). Separation was performed on a C18 reversed-phase column with a gradient of water and methanol containing 0.1% formic acid. The method was validated to meet a specification limit of 15 ng/mg, with a linear range of 0.5–80 ng/mL, effectively covering 10–400% of the regulatory threshold. The method exhibited an excellent performance in terms of specificity, accuracy, precision, linearity, and robustness. It achieved a limit of detection (LOD) of 0.2 ng/mL (0.30 ng/mg) and a limit of quantification (LOQ) of 0.5 ng/mL (0.75 ng/mg), alongside a comprehensive uncertainty analysis with an expanded uncertainty of ±3.86 mg/kg. Application to commercial atenolol products confirmed the reliability and practical utility of the method. This validated approach offers a critical tool for pharmaceutical manufacturers and regulatory agencies to monitor and control N-nitroso-atenolol, ensuring compliance and enhancing patient safety. |
| format | Article |
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| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-1f221bb1128a40ef8ed9ef8fc1093ed72025-08-20T01:56:45ZengMDPI AGSeparations2297-87392025-05-0112512210.3390/separations12050122Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based PharmaceuticalsSoonho Kwon0Sang-Hyun Ahn1Yongha Chang2Joon-Sang Park3Hwangeui Cho4Jung-Bok Kim5Corporation VIVAGEN, Seongnam 13219, Republic of KoreaCorporation VIVAGEN, Seongnam 13219, Republic of KoreaCorporation VIVAGEN, Seongnam 13219, Republic of KoreaCorporation VIVAGEN, Seongnam 13219, Republic of KoreaInstitute of New Drug Development, School of Pharmacy, Jeonbuk National University, Jeonju 54896, Republic of KoreaDepartment of Yuhan Biotechnology, School of Bio-Health Sciences, Yuhan University, Bucheon 14780, Republic of KoreaThe recent detection of N-nitroso-atenolol, a mutagenic and potentially carcinogenic impurity in atenolol-based pharmaceuticals, has raised serious safety concerns and emphasized the need for stringent analytical control. This study developed and validated a highly sensitive LC-MS/MS method for quantifying N-nitroso-atenolol in both active pharmaceutical ingredients (APIs) and finished products. Quantification was carried out using multiple reaction monitoring (MRM) under positive-mode electrospray ionization (ESI). Separation was performed on a C18 reversed-phase column with a gradient of water and methanol containing 0.1% formic acid. The method was validated to meet a specification limit of 15 ng/mg, with a linear range of 0.5–80 ng/mL, effectively covering 10–400% of the regulatory threshold. The method exhibited an excellent performance in terms of specificity, accuracy, precision, linearity, and robustness. It achieved a limit of detection (LOD) of 0.2 ng/mL (0.30 ng/mg) and a limit of quantification (LOQ) of 0.5 ng/mL (0.75 ng/mg), alongside a comprehensive uncertainty analysis with an expanded uncertainty of ±3.86 mg/kg. Application to commercial atenolol products confirmed the reliability and practical utility of the method. This validated approach offers a critical tool for pharmaceutical manufacturers and regulatory agencies to monitor and control N-nitroso-atenolol, ensuring compliance and enhancing patient safety.https://www.mdpi.com/2297-8739/12/5/122N-nitroso-atenololNDSRIsLC-MS/MS |
| spellingShingle | Soonho Kwon Sang-Hyun Ahn Yongha Chang Joon-Sang Park Hwangeui Cho Jung-Bok Kim Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals Separations N-nitroso-atenolol NDSRIs LC-MS/MS |
| title | Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals |
| title_full | Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals |
| title_fullStr | Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals |
| title_full_unstemmed | Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals |
| title_short | Development and Validation of a Sensitive LC-MS/MS Method for the Determination of N-Nitroso-Atenolol in Atenolol-Based Pharmaceuticals |
| title_sort | development and validation of a sensitive lc ms ms method for the determination of n nitroso atenolol in atenolol based pharmaceuticals |
| topic | N-nitroso-atenolol NDSRIs LC-MS/MS |
| url | https://www.mdpi.com/2297-8739/12/5/122 |
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