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Development and Validation of LC–MS/MS Method for the Quantitation of Infliximab in Human Serum

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Abstract

Liquid chromatography tandem mass spectrometry (LC–MS/MS), especially triple-quadrupole mass spectrometry, has become an attractive alternative method to ligand binding assays for therapeutic monoclonal antibody quantification in biological samples, but the use of an internal standard with infliximab LC–MS/MS assays has not been reported yet. In this study, an improved LC–MS/MS method for quantification of infliximab in human serum was developed and validated. A surrogate peptide was used as a representative of infliximab which was cleaved for the quantification of infliximab based on LC–MS/MS assay. A stable isotope-labeled signature peptide was used as the internal standard (IS). The results showed linearity in the range of 0.39–100 μg mL−1; the lower limit of quantification (LLOQ), and the lower limit of detection were 0.39 and 0.0975 μg mL−1, respectively. The quality control (QC) data showed that the within-run, between-run precision (%RSD) and accuracy (%RE) conformed to the acceptance criteria of ±15 % for calibration standards and QCs (±20 % at the LLOQ). Other validation parameters including selectivity, methanol precipitation efficiency, serum matrix effect, stability, and auto-sampler carry-over were also evaluated. This improved LC–MS/MS method might be a promising LC–MS-based methodology for pharmacokinetic studies of other recombinant monoclonal antibodies.

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Acknowledgments

We thank Waters Company Laboratories for cooperation with us. This work was supported by grants from the Natural Science Foundation of China (81330061), Ministry of Science and Technology of China (973 projects 2010CB833605 and 863 projects 2014AA021004), State Key Project for New Drug Development (2013ZX09101021; 2013ZX09401303), Shanghai Commission of Science and Technology (Key Laboratory and Projects 13DZ1930100), and Shanghai Excellent technical leader (13XD1424000).

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

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    Authors and Affiliations

    1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, China

      Xiaoyun Peng & Boning Liu

    2. International Joint Cancer Institute, Second Military Medical University, Shanghai, China

      Xiaoyun Peng, Boning Liu, Yantao Li, Qingcheng Guo, Hao Wang, Dapeng Zhang, Jianxin Dai, Sheng Hou & Yajun Guo

    3. State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai Key Laboratory of Cell Engineering, Shanghai, China

      Xiaoyun Peng, Boning Liu, Yantao Li, Huaizu Guo, Qingcheng Guo, Jin Xu, Hao Wang, Dapeng Zhang, Jianxin Dai, Sheng Hou & Yajun Guo

    4. School of Pharmacy, Liaocheng University, Liaocheng, China

      Yajun Guo

    5. Waters Corporation, Shanghai, China

      Hui Wang & Xi Chen

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    1. Xiaoyun Peng
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    2. Boning Liu
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    Corresponding author

    Correspondence to Sheng Hou.

    Additional information

    X. Peng, B. Liu, Y. Li were contributed equally to this work.

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    Peng, X., Liu, B., Li, Y. et al. Development and Validation of LC–MS/MS Method for the Quantitation of Infliximab in Human Serum. Chromatographia 78, 521–531 (2015). https://doi.org/10.1007/s10337-015-2866-2

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    • DOI: https://doi.org/10.1007/s10337-015-2866-2

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