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Variable VOCs in plastic culture flasks and their potential impact on cell volatile biomarkers

Analytical and Bioanalytical Chemistry volume 412, pages 5397–5408 (2020)Cite this article

Abstract

In order to find out cancer markers in human breath, in vitro cell culture is often used to study the characteristic volatile organic compounds (VOCs). In the cell culture process, disposable vessels are frequently adopted. However, these vessels are normally made of plastic, and they have the possibility to release some VOCs, which may interfere with the cell-specific volatiles and even can result in an incorrect conclusion. In this study, by using glass cell culture flasks as control, the headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS) analyses of the VOCs in plastic cell culture flasks were systematically carried out for the first time. A total of 35 VOCs were detected in five brands of flasks. In each flask, there were between 13 and 25 volatile compounds. Furthermore, the components and packaging bag of each flask were also sampled and analyzed by HS-SPME-GC-MS. The results show that the flask cap, septum, flask body, and packaging bag exhibit respectively different volatile behaviors. The former two parts release the most volatiles which have obvious contributions to the headspace gases in the flasks, while the flask body mainly liberates styrene. For different flasks packed within the same bag, the headspace analyses show that their residual VOCs are inconsistent with each other. Moreover, the residual VOCs in the same flask are variable in three consecutive days. These results indicate that the multiple flasks in parallel cell culture experiments, or the same flask with different cell culture durations, will produce an indelible disturbance to the cell-specific VOCs. In addition, among the 35 VOCs detectable in five brands of empty plastic flasks, 15 VOCs were previously reported as characteristic VOCs from lung cancer, melanoma, cervical cancer cells, or normal cells. This is an alert that, when using plastic flasks, it must be careful to treat the possible interference from the background VOCs in the flasks. This study demonstrates that the cell culture tool needs to be standardized, and the clean glass or metal vessels are strongly recommended for usage when studying cell volatile biomarkers.

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Funding

This research was funded by the National Natural Science Foundation of China (21876176, 21705152); the Youth Innovation Promotion Association, Chinese Academy of Sciences (2019432); the Key Program of 13th Five-Year Plan, Hefei Institutes of Physical Science, Chinese Academy of Sciences (KP-2017-25); and the joint fund between the Second Affiliated Hospital of Anhui Medical University and the Center of Medical Physics and Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences (LHJJ2020006).

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Affiliations

  1. Anhui Province Key Laboratory of Medical Physics and Technology, Institute of Health & Medical Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, 230031, Anhui, China

    Yajing Chu, Jijuan Zhou, Dianlong Ge, Yan Lu, Xue Zou, Lei Xia, Chaoqun Huang, Chengyin Shen & Yannan Chu

  2. Institute of Physical Science and Information Technology, Anhui University, Hefei, 230601, Anhui, China

    Yajing Chu

  3. University of Science and Technology of China, Hefei, 230026, Anhui, China

    Jijuan Zhou & Dianlong Ge

  4. Hefei Cancer Hospital, Chinese Academy of Sciences, Hefei, 230031, Anhui, China

    Yan Lu & Chengyin Shen

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  1. Yajing Chu
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Correspondence to Yannan Chu.

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Chu, Y., Zhou, J., Ge, D. et al. Variable VOCs in plastic culture flasks and their potential impact on cell volatile biomarkers. Anal Bioanal Chem 412, 5397–5408 (2020). https://doi.org/10.1007/s00216-020-02756-9

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  • DOI: https://doi.org/10.1007/s00216-020-02756-9

Keywords

  • Culture flask
  • Cancer cell
  • VOCs
  • Biomarker
  • GC-MS