Validated GC–APCI–MS/MS Method for Sensitive Determination of N‑Nitrosodimethylamine in Valsartan Hydrochloride

The detection of trace-level nitrosamine impurities remains a critical challenge in pharmaceutical analysis, requiring highly selective and reproducible chromatographic and spectroscopic techniques. In this study, we present a validated gas chromatography–atmospheric pressure chemical ionization tandem mass spectrometry (GC–APCI–MS/MS) method for the quantification of N-nitrosodimethylamine (NDMA) in valsartan hydrochloride. The method eliminates pre-concentration steps, enabling rapid high-throughput analysis with a runtime of ~12 minutes. Validation was performed in accordance with ICH Q2(R1) guidelines. The method demonstrated excellent linearity (R² = 0.99985), low detection limit (LOD = 0.045 ppm), quantitation limit (LOQ = 0.090 ppm), and high recovery (90.4–97.5%). Compared with conventional electron ionization (EI) approaches, APCI provided enhanced selectivity and reduced matrix interference. The robustness of the method was confirmed through precision studies across multiple concentration levels, with %RSD values well within acceptance criteria. This work highlights the analytical advantages of GC–APCI–MS/MS for nitrosamine determination and establishes a reliable platform for pharmaceutical quality control, with potential extension to environmental and food safety applications.

Gas Chromatography, Genotoxic Impurities, Mass Spectrometry, NDMA, Valsartan Hydrochloride

1. International Council for Harmonisation (ICH). ICH guideline M7(R1) on assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk. ICH Harmonised Tripartite Guideline. 2015. [Google Scholar] [Crossref]

2. IARC Working Group. IARC monographs on the evaluation of carcinogenic risks to humans: Ingested nitrate and nitrite, and cyanobacterial peptide toxins. IARC Monogr Eval Carcinog Risks Hum. 2010;94:1–412. [Google Scholar] [Crossref]

3. Al-Sabti B, Harbali J. Development and validation of an analytical method for quantitative determination of three potentially genotoxic impurities in vildagliptin drug material using HPLC–MS. J Sep Sci. 2021;44:2587–95. [Google Scholar] [Crossref]

4. Prada F, Stashenko EE, Martinez JR. LC/MS study of the diversity and distribution of pyrrolizidine alkaloids in Crotalaria species growing in Colombia. J Sep Sci. 2020;43:4322–37. [Google Scholar] [Crossref]

5. Tang Y, Zhang JL. Recent developments in DNA adduct analysis using liquid chromatography coupled with mass spectrometry. J Sep Sci. 2020;43:31–55. [Google Scholar] [Crossref]

6. Yahagi T, Nagao M, Seino Y, Matsushima T, Sugimura T. Mutagenicities of N-nitrosamines on Salmonella. Mutat Res. 1977;48:121–9. [Google Scholar] [Crossref]

7. Lopez-Rodriguez R, McManus JA, Murphy NS, Ott MA, Burns MJ. Pathways for N-nitroso compound formation: Secondary amines and beyond. Org Process Res Dev. 2020;24:1558–65. [Google Scholar] [Crossref]

8. Parr MK, Joseph JF. NDMA impurity in valsartan and other pharmaceutical products: Analytical methods for the determination of N-nitrosamines. J Pharm Biomed Anal. 2019;164:536–49. [Google Scholar] [Crossref]

9. Li Z, Wang J, Chen X, Hu S, Gong T, Xian Q. A novel molecularly imprinted polymer–solid phase extraction method coupled with high performance liquid chromatography tandem mass spectrometry for the determination of nitrosamines in water and beverage samples. Food Chem. 2019;292:267–74 [Google Scholar] [Crossref]

10. Alshehri YM, Alghamdi TS, Aldawsari FS. HS-SPME-GC-MS as an alternative method for NDMA analysis in ranitidine products. J Pharm Biomed Anal. 2020;191:113582. [Google Scholar] [Crossref]

11. Schmidtsdorff S, Schmidt AH. Simultaneous detection of nitrosamines and other sartan-related impurities in active pharmaceutical ingredients by supercritical fluid chromatography. J Pharm Biomed Anal. 2019;174:151–60. [Google Scholar] [Crossref]

12. Soergel F, Kinzig M, Abdel-Tawab M, Bidmon C, Schreiber A, Ermel S, et al. The contamination of valsartan and other sartans, part 1: New findings. J Pharm Biomed Anal. 2019;172:395–405. [Google Scholar] [Crossref]

13. Schmidtsdorff S, Schmidt AH. Simultaneous detection of nitrosamines and other sartan-related impurities in active pharmaceutical ingredients by supercritical fluid chromatography. J Pharm Biomed Anal. 2019;174:151–60. [Google Scholar] [Crossref]

14. Fritzsche M, Blom G, Keitel J, Goettsche A, Seegel M, Leicht S, et al. NDMA analytics in metformin products: Comparison of methods and pitfalls. Eur J Pharm Sci. 2022;168:106026. [Google Scholar] [Crossref]

15. Roback SL, Kodamatani H, Fujioka T, Plumlee MH. Validation of a novel direct-injection chemiluminescence-based method for N-nitrosamine analysis in advanced-treated recycled water, drinking water, and wastewater. Environ Sci Water Res Technol. 2020;6:1106–15. [Google Scholar] [Crossref]

16. Schmidtsdorff S, Neumann J, Schmidt AH, Parr MK. Analytical lifecycle management for comprehensive and universal nitrosamine analysis in various pharmaceutical formulations by supercritical fluid chromatography. J Pharm Biomed Anal. 2021;197:113960. [Google Scholar] [Crossref]

17. Bian Y, Zhang Y, Zhou Y, Li GH, Feng XS. Progress in the pretreatment and analysis of N-nitrosamines: An update since 2010. Crit Rev Food Sci Nutr. 2021;61:3626–60. [Google Scholar] [Crossref]

18. Yahaya A, Babatunde D, Olaniyan LWB, Agboola O. Application of chromatographic techniques in the analysis of total nitrosamines in water. Heliyon. 2020;6:e03447. [Google Scholar] [Crossref]

19. Zan K, Hu X, Li Y, Wang Y, Jin H, Zuo T, et al. Simultaneous determination of eight pyrrolizidine alkaloids in various parts of Eupatorium lindleyanum by ultra high performance liquid chromatography tandem mass spectrometry and risk assessments based on a real-life exposure scenario. J Sep Sci. 2021;44:3237–47. [Google Scholar] [Crossref]

20. Li W, Cang S, Sun Z, Bi K, Li Q, Li Z, et al. Development of an LC-MS/MS method for simultaneous quantitative analysis of macromolecular pharmaceutical adjuvant 2-hydroxypropyl beta-cyclodextrin and active pharmaceutical ingredients butylphthalide in rat plasma. J Sep Sci. 2021;44:2680–92. [Google Scholar] [Crossref]

21. Lashgari M, Yamini Y, Basheer C, Lee HK. Ordered mesoporous carbon as sorbent for the extraction of N-nitrosamines in wastewater and swimming pool water. J Chromatogr A. 2017;1513:35–41. [Google Scholar] [Crossref]

22. Lu S, Wu D, Li G, Lv Z, Gong P, Xia L, et al. Facile and sensitive determination of N-nitrosamines in food samples by high-performance liquid chromatography via combining fluorescent labeling with dispersive liquid–liquid microextraction. Food Chem. 2017;234:408–15. [Google Scholar] [Crossref]

23. Amelin VG, Bol’shakov DS. Rapid identification and determination of N-nitrosamines in food products by ultra high-performance liquid chromatography–high resolution quadrupole-time-of-flight mass spectrometry by exact masses of protonated molecules. J Anal Chem. 2019;74:39–46. [Google Scholar] [Crossref]

24. Liu J, Xie B, Mai B, Cai Q, He R, Guo D, et al. Development of a sensitive and stable GC-MS/MS method for simultaneous determination of four N-nitrosamine genotoxic impurities in sartan substances. J Anal Sci Technol. 2021;12:3. [Google Scholar] [Crossref]

25. Wang X, Qin Y, Nie C, Guo J, Pan L, Xie F, et al. Smokeless tobacco analysis: Simultaneous extraction and purification of alkaloids, volatile N-nitrosamines, and polycyclic hydrocarbons for GC-MS/MS. J Sep Sci. 2021;44:2642–54. [Google Scholar] [Crossref]

26. Zheng JJ, Kirkpatrick CL, Lee D, Han XX, Martinez AI, Gallagher K, et al. A full evaporation static headspace gas chromatography method with nitrogen phosphorus detection for ultrasensitive analysis of semi-volatile nitrosamines in pharmaceutical products. AAPS J. 2022;24:23. [Google Scholar] [Crossref]

27. Roasa J, Liu H, Shao S. An optimised HS-SPME-GC-MS method for the detection of volatile nitrosamines in meat samples. Food Addit Contam Part A. 2019;36:396–404. [Google Scholar] [Crossref]

28. Amelin VG, Lavrukhin DK. Combination of microwave heating extraction and dispersive liquid-liquid microextraction for the determination of nitrosoamines in foods using gas–liquid chromatography with a mass-spectrometric detector. J Anal Chem. 2016;71:359–64. [Google Scholar] [Crossref]

29. Chen X, Yang S, Hu Q, Cheng H, Chen Z, Ouyang G. Efficient sampling and determination of airborne N-nitrosamines by needle trap device coupled with gas chromatography–mass spectrometry. Microchem J. 2018;139:480–6. [Google Scholar] [Crossref]

30. Zhang Q, Jin L, Zhang F, Yao K, Ren Y, Zhang J, et al. Analysis of 7 volatile N-nitrosamines in Chinese Sichuan salted vegetables by gas chromatography–tandem mass spectrometry coupled to modified QuEchERS extraction. Food Control. 2019;98:342–7. [Google Scholar] [Crossref]

31. Jiang PT, Wu KL, Wang HT, Chen SF. The use of atmospheric pressure chemical ionization for pesticide analysis using liquid chromatography mass spectrometry. J Food Drug Anal. 2022;30:163–71. [Google Scholar] [Crossref]

32. Zhou H, Huang Q, Wu XL, Zhan BD, Chen DY, Lei MK, et al. Rapid and selective determination of 9 nitrosamines in biological samples using ultra-high performance liquid chromatography–triple quadrupole linear ion trap mass spectrometry. Anal Methods. 2022;14:2249–54. [Google Scholar] [Crossref]

33. Solanki R, Patel C, Patel R, Maradiya A, Parikh S, Gayakvad B. UHPLC-APCI-TQ-MS analytical method to quantify nitrosamine impurities in the commercial formulation of metformin and glipizide. J Liq Chromatogr Relat Technol. 2022. doi:10.1080/10826076.2022.2126856. [Google Scholar] [Crossref]

34. Andrade JM, Gomez-Carracedo MP. Notes on the use of Mandel’s test to check for nonlinearity in laboratory calibrations. Anal Methods. 2013;5:1145–9. [Google Scholar] [Crossref]

35. Yin M, Hu Y, Fan H, Wang Q, Wang M, Wang W, Shi C. Method for trace determination of N-nitrosamine impurities in metronidazole benzoate using high-performance liquid chromatography coupled with atmospheric-pressure chemical ionization tandem mass spectrometry. J Sep Sci. 2023;46:e2200225. doi:10.1002/jssc.202200225. [Google Scholar] [Crossref]

• Assessment of the Role of Artificial Intelligence in Repositioning TVET for Economic Development in Nigeria
• Teachers’ Use of Assure Model Instructional Design on Learners’ Problem Solving Efficacy in Secondary Schools in Bungoma County, Kenya
• “E-Booksan Ang Kaalaman”: Development, Validation, and Utilization of Electronic Book in Academic Performance of Grade 9 Students in Social Studies
• Analyzing EFL University Students’ Academic Speaking Skills Through Self-Recorded Video Presentation
• Major Findings of The Study on Total Quality Management in Teachers’ Education Institutions (TEIs) In Assam – An Evaluative Study