A Robust and Universal LC–MS/MS Method for Determination of N-Nitrosodimethylamine in Pharmaceuticals Using a C30 Column -Pub

A Robust and Universal LC–MS/MS Method for Determination of N-Nitrosodimethylamine in Pharmaceuticals Using a C30 Column

Abstract:
Since the 2018 valsartan recall, the genotoxic impurity N- nitrosodimethylamine (NDMA) has been frequently detected in various pharmaceuticals. Matrix variability often complicates routine testing, requiring customized analyses. We developed a universal LC–MS/MS method enabling consistent NDMA detection across diverse pharmaceuticals. Separation utilized a C30 column (150 mm × 4.6 mm I.D., 5 µm), offering superior retention and shape selectivity for polar nitrosamines compared to conventional reversed phases. Unlike C18 or pentafluorophenyl phases, which often exhibit limited retention for NDMA, the C30 phase provides exceptional shape selectivity and enhanced hydrophobic interactions. This structural advantage, supplemented by its resistance to phase dewetting under highly aqueous conditions further ensures robust resolution between NDMA and complex drug matrices. The mobile phase consisted of 0.1% v/v formic acid in water and methanol under gradient elution at 1.0 mL/min. Detection used atmospheric pressure chemical ionization in positive ion mode. The method demonstrated a linear range of 2.0–100.0 ng/mL (r 2 > 0.999), with a limit of detection of 1.0 ng/mL and a limit of quantification of 2.0 ng/mL. Validation per International Council for Harmonisation (ICH) guidelines confirmed accuracy (87.7%–115.5%) and precision (coefficient of variation, CV ≤ 10.7%). Application to 30 diverse pharmaceutical products, including sartans and ranitidine, showed robust resolution (> 2.0) between the analyte and active pharmaceutical ingredients (APIs). Notably, NDMA was quantified in historical batches of ranitidine (164.83 ± 5.68 ng/mL), nizatidine (10.25 ± 0.52 ng/mL), and amitriptyline (2.28 ± 0.19 ng/mL). While the histamine type 2 receptor antagonists significantly exceeded the acceptable daily intake limit, the remaining 27 products showed no detectable NDMA. These findings highlight the method’s effectiveness for real-world surveillance and the critical risk of post-manufacturing NDMA generation during prolonged storage. This universal C30-based method provides a practical, reliable tool for routine screening, facilitating regulatory compliance and improved patient safety without drugspecific method development.

A Robust and Universal LC–MS/MS Method for Determination of N-Nitrosodimethylamine in Pharmaceuticals Using a C30 Column

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Routine NDMA testing often requires drug-specific methods due to matrix variability and complex active pharmaceutical ingredients (APIs). This study presents a universal LC-APCI-MS/MS approach using a C30 column, successfully separating trace NDMA from 30 diverse APIs without the need for complex, time-consuming sample preparation.

Key Findings:

  • C30 stationary phases offer superior shape selectivity and resist phase dewetting in highly aqueous conditions.
  • The method achieved a 2.0 ng/mL limit of quantification for the majority of the tested APIs.
  • Atmospheric pressure chemical ionization (APCI) was utilized over electrospray ionization (ESI) to minimize matrix ion suppression.
  • High NDMA concentrations were quantified in historical, aged batches of ranitidine, nizatidine, and amitriptyline.

Lessons for the Community:

  • Strategic use of C30 columns can eliminate extensive drug-specific method redevelopment for polar nitrosamines.
  • “Dilute-and-shoot” methods are highly viable if the chromatographic column offers sufficient retention and selectivity.
  • Implementing a divert valve is crucial to protect the mass spectrometer from massive API overload.
  • Aged, structurally vulnerable APIs remain a significant risk for post-manufacturing NDMA generation during prolonged storage.
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