Research On Detection Method Of Nitrosamine Impurities In Pharmaceuticals

Genotoxic impurities can cause chromosome breakage and rearrangement at small levels,change genetic material,and cause the risk of teratogenic and cancerous transformation.Therefore,the control of genotoxic impurities in drugs has become the focus of attention.Nitrosamine impurities(NAs)are one of these categories and belong to the“cohort of concern” mentioned in the ICH M7(R1)Guideline,Assessment and Control of DNA Reactive(Mutogenic)Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk.With the N-nitrosamine compounds like N-nitrosodimethylamine(NDMA),Nnitrosamine diethylamine(NDEA)and N-nitroso-N-Methyl-4-aminobutyric acid(NMBA)being detected successively in commonly used drugs such as valsartan,metformin and ranitidine,domestic and foreign drug regulatory agencies and manufacturers are paying more and more attention to the risk of contamination of nitrosamine compounds in drugs.Accurate and sensitive determination of the type and content of nitrosamine impurities in drugs,and controlling the content of nitrosamine impurities below the acceptable limit,is the difficulty and focus of ensuring drug safety.The combined technology of mass spectrometry and gas chromatography or liquid chromatography is a reliable technique for the detection of trace impurities in drugs.Chromatography technology uses the powerful separation ability of the chromatographic column to separate the compounds to be detected from other interfering matrices,the high sensitivity and high selectivity of mass spectrometry enables accurate qualitative and quantitative determination of trace substances.Therefore,it is widely used in the detection of impurities,especially the combination of chromatography and triple quadrupole mass spectrometry(or quadrupole mass spectrometry)is the most commonly used quantitative detection method.In addition,quadrupole/electrostatic field orbitrap mass spectrometers are increasingly being applied to the detection of trace impurities in drugs due to their high sensitivity and high resolution,providing another detection method for the detection of NAs.In this paper,a new method for detecting the N-nitroamine impurities in drugs based on combination of chromatography and mass spectrometry is developed.The contents are as follows:1.An overview of nitrosamine compounds,mainly including the toxicity of nitrosamine compounds,the way in which they are produced in drugs,the detection methods previously issued by regulatory agencies around the world,and the control standards and limits of nitrosamine impurities in drugs was introduced.Meanwhile,the mass spectrometry ion source and analyzer suitable for the detection of trace impurities were also systematically introduced and the specific content and significance of this research were also clarified.2.In view of the problem that most of the current detection methods can only detect the most common NDMA and NDEA in drugs while cannot guarantee that there are no other NAs in drugs,this study has developed GC-MS and GC-MS/MS methods for detection of a variety of NAs which have been confirmed that may be present in drugs were validated with sartan drugs.In this part,the influence of sample extraction solvent and the difference in sensitivity and specificity of GC-MS and GC-MS/MS in the detection of multiple NAs were also validated.The detection provides a reference method for detection of the common nitrosamine impurities in drugs.3.First of all,for tidine drugs(ranitidine and nizatidine)and metformin raw materials and preparations,the LC-MS/MS method for the detection of NDMA in the two drugs was developed in the country for the first time.Due to the strong operability and repeatability of the method,it was recommended to domestic regulatory agencies in a timely manner and helpful for the National Medical Products Administration to grasp the quality of domestic drugs.Secondly,aiming at the problem that the polar nitrosamine compounds such as NMBA are difficult to be directly detected by GC-MS,as well as other nitrosamine impurities that may be present in drugs reported at home and abroad,In this study,an LC-MS/MS method was also established for the simultaneous detection of seven nitrosamines in drugs.The method verification results show that it can meet the expected use.In addition,ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry is increasingly used in the field of trace impurities.Therefore,the UPLC-QE HRMS method was also established for the detection of various NAs in drugs,which enrich the diversity of detection methods for nitrosamine impurities.Finally,considering the possibility of other nitrosamine impurities in drugs,an LC-MS/MS method was also established in this study to detect 16 common nitrosamine impurities in drugs.Meanwhile,the drug Telmisartan was verified via the method and 6 batches of the samples were tested.This method can not only detect nitrosamine impurities in sartan drugs,but also can be used as a screening method for unknown nitrosamine impurities in drugs.The purpose of this study is to detect and quantify nitrosamine impurities in drugs,using GC-MS and HPLC methods in tandem with triple quadrupole mass spectrometry and quadrupole/electrostatic field orbitrap mass spectrometry,covering common nitrosamines in drugs Amine impurities.It can ensure the quality and safety of drugs and the medication safety,and provide a useful reference for accelerating the construction of the most stringent drug regulatory system.