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A Simple and Sensitive UPLC–MS/MS Method for the Simultaneous and Trace Level Quantification of Two Potential Genotoxic Impurities in Midazolam Drug Substance

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Abstract

To promote the quality control of midazolam and ensure the life safety of patients, a sensitive and simple ultra-high performance liquid chromatography–mass spectrometry (UPLC–MS/MS) method was developed for the trace determination of two potential genotoxic impurities (PGIs) in midazolam raw materials under the positive electrospray ionization (ESI) mode with multiple reaction monitoring (MRM). The two PGIs were well separated in a gradient elution mode at a 0.4 mL/min flow rate on an ACQUITY UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 µm) that was maintained at 40 °C. 0.01 mol/L ammonium formate aqueous solution (0.1% formic acid) and acetonitrile (0.1% formic acid) were mobile phase A and mobile phase B, respectively. The m/z of 343.00/123.00 for PGI-1and m/z of 331.20/301.20 for PGI-2 were detected in a triple quadrupole mass detector. The calibration curves showed that the two PGIs have good linearity between the concentration range of 0.004 and 0.8 ppm. The correlation coefficients of PGI-1 and PGI-2 were 0.9996 and 0.9998, respectively. The recoveries of the two PGIs at three concentration levels were in the range of 81–103%. The limits of quantification (LOQ) for PGI-1 and PGI-2 were 0.0545 and 0.0111 ng/mL respectively. The precision relative standard deviation (RSD) of each impurity was observed less than 3.0%. Method validation results indicated that the linearity, accuracy, precision, and sensitivity of the developed UPLC–MS/MS method are satisfactory. Thus, the method can quantitatively detect the two trace PGIs in midazolam. This study can provide a reference for manufacturers and relevant researchers of midazolam, reduce the quality risk of midazolam drugs and ensure the safety of patients.

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Important data generated or analyzed during this study are included in this published article and its supplementary information file.

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Acknowledgements

This work was supported by Chongqing Plan for High Level Talents project of CQ CSTC (No.: cstc2021ycjh-bgzxm0315) and the National Major Scientific and Technological Special Project of the Ministry of Science and Technology of China (2017ZX09101001).

Funding

Chongqing Plan for High Level Talents project of CQ CSTC (No.: cstc2021ycjh-bgzxm0315); the National Major Scientific and Technological Special Project of the Ministry of Science and Technology of China (2017ZX09101001).

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Authors and Affiliations

  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400044, China

    Dandan Chen, Li Huang & Jianhua Wang

  2. Chemistry Laboratory, Chongqing Institute for Food and Drug Control, Chongqing, 401121, China

    Lijun Luo, Dandan Shen & Linggao Zeng

  3. NMPA Key Laboratory for Quality Monitoring of Narcotic Drugs and Psychotropic Substances, Chongqing, 401121, China

    Lijun Luo, Dandan Shen & Linggao Zeng

Authors
  1. Dandan Chen
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  2. Lijun Luo
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  4. Li Huang
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  5. Linggao Zeng
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  6. Jianhua Wang
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Contributions

The first draft of the manuscript was written by DC. The experimental protocol was formulated by DC and LL. Material and reagent preparation were performed by DC. Data collection and analysis were done by DC, DS and LH. Writing guidance and advice were provided by JW and LZ. All authors read and approved the final manuscript.

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Correspondence to Linggao Zeng or Jianhua Wang.

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Chen, D., Luo, L., Shen, D. et al. A Simple and Sensitive UPLC–MS/MS Method for the Simultaneous and Trace Level Quantification of Two Potential Genotoxic Impurities in Midazolam Drug Substance. Chromatographia 86, 167–173 (2023). https://doi.org/10.1007/s10337-022-04221-4

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