Trace Level Quantification of 4-Methyl-1-nitrosopiperazin in Rifampicin Capsules by LC-MS/MS

Abstract

Rifampicin is a first-line anti-tuberculosis drug. However, in August 2020, the presence of 1-methyl-4-nitrosopiperazine (MNP), a nitrosamine impurity, was detected by the United Stated Food and Drug Administration (US FDA) in rifampicin capsules. Consequently, the development of efficient methods for the detection of MNP is an important objective. In this study, the MNP present in rifampicin capsules was detected using LC-MS/MS. A total of 27 batches from nine manufacturers in the Chinese market were tested, with MNP (0.33–2.36 ppm) being detected in all samples at levels exceeding the maximum acceptable intake limit of 0.16 ppm initially set by the FDA. However, after considering the associated benefits and risks, the FDA-approved limit was revised to 5 ppm; hence, all the samples examined herein exhibited MNP levels well below the required limit. Furthermore, the results of forced degradation experiments suggest that MNP is formed by the thermal degradation of rifampicin.

A New Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method to Monitor 1-Amino-4-Methyl-Piperazine and 1-Methyl-4-Nitroso-Piperazine in Rifampicin Hydrolysis: A Standard Addition Approach to Quantify Nitrosamine in Rifampicin

“…As shown, the standard addition approach gave very good results for all validation parameters. This is, reasonably, the best approach to quantify MNP in RIF because MNP is always present in the matrix and because the matrix effect is not negligible, although the RIF concentration used in this study was three times lower than previously used [10][11][12][13]. This method will be extended to the analysis of drug products where the matrix effect is usually higher.…”

“…As an alternative approach to the single point analysis (limit test) published by the FDA and to the previously proposed external calibration [6,[10][11][12][13], a standard addition method was used for the analysis of MNP in RIF in drug substances. This approach was preferred because all the RIF matrices analyzed contained MNP and the matrix effect cannot be ignored at the RIF concentration used for MNP analysis, which is 30 mg mL −1 in the FDA method and 27 mg mL −1 in recent studies, respectively [11][12][13]. In addition, this approach is more accurate than a limit test to quantify MNP when the concentration of MNP is required rather than simply meeting the 5 ppm limit.…”

“…This is a single-point test (limit test) and the MNP concentration in the sample is calculated based on a single-concentration solution. Few other papers have been published on the analysis of MNP in RIF, but to the best of our knowledge, there are no LC-MS methods available in the literature for the quantification of AMP [6,[11][12][13].…”

Determination of Aflatoxin B1 in Grains by Aptamer Affinity Column Enrichment and Purification Coupled with High Performance Liquid Chromatography Detection

“…Thus far, analytical methods based on chromatography, spectroscopy, and immunoassay have been developed and used to determine AFB 1 . The chromatographic methods include high performance liquid chromatography (HPLC) [ 9 ], thin-layer chromatography (TLC) [ 10 ], and liquid chromatography-mass spectrometry (LC-MS) [ 11 ]. The spectroscopic methods include fluorescence spectrophotometry [ 12 ], infrared (IR) spectroscopy [ 13 ], and chromatin interacting protein-mass spectrometry (Chip-MS) [ 14 ].…”

Development of a Sensitive Screening Method for Simultaneous Determination of Nine Genotoxic Nitrosamines in Active Pharmaceutical Ingredients by GC-MS

“…Since then, widespread investigations by regulatory agencies, including the European Medicines Agency (EMA) and United States Food and Drug Administration (US FDA), were undertaken. In drugs such as angiotensin II receptor blockers (ARBs), ranitidine, metformin, rifampin, rifapentine, and, recently, verenicline, N -nitrosodimethylamine (NDMA) and other nitrosamines have been detected [ 2 , 3 ]. Nitrosamines are known as probable human carcinogens (IARC 2A group for NDMA), potent genotoxic agents, and “cohort of concern” compounds according to ICH M7 guidance [ 4 , 5 ].…”