Elemental Analysis of Chemically Defined Cell Culture Media by ICP-MS

Importance of the Topic

Chemically defined cell culture media (CDM) are critical for the reproducible production of biopharmaceuticals, vaccines and advanced cell therapies. Consistent metal ion composition in CDM supports enzyme cofactors and cell signaling pathways, directly impacting product yield, quality and safety. Controlling trace and major elements minimizes batch-to-batch variability, microbial contamination and regulatory concerns about elemental impurities.

Objectives and Overview of the Study

This work presents a multi-element analytical method using the Agilent 7900 ICP-MS to quantify major and trace elements in CDM. Two serum-free media—Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F-12—were selected to compare element profiles across different lots and brands. The goals were method development, optimization, stability testing and demonstration of accuracy for quality control of CDM.

Methodology and Instrumentation

Samples were prepared by ten-fold dilution in 2 % HNO₃. Calibration standards covered 0.005–40 ppm for major elements and 0.05–400 ppb for trace elements, with QC standards at 10 ppm and 100 ppb. A single-point internal standard (ISTD) was added online. Analytical sequence included initial calibration, verification, sample analysis with spiked recovery blocks and continuing calibration verifications every ten samples.

Used Instrumentation

• Agilent 7900 ICP-MS equipped with ORS4 collision/reaction cell in helium (He) mode for polyatomic interference removal
• Nickel sampling and skimmer cones; quartz spray chamber at 2 °C; MicroMist concentric nebulizer; quartz torch with 2.5 mm injector
• Agilent I-AS autosampler; MassHunter General Purpose preset method with autotuned plasma and lens parameters

Results and Discussion

• Detection limits ranged from 0.0001 ppm (Fe) to 0.3416 ppb (Al), with linear calibration (R > 0.999) across all elements.
• Spike recoveries for major and trace elements fell between 95 % and 115 %, demonstrating quantitative accuracy in fortified DMEM and Ham’s F-12.
• Internal standard and continuing calibration verification signals remained within ±20 % and ±10 %, respectively, over eight-hour runs, confirming long-term stability and matrix tolerance.
• Major element profiles of DMEM were consistent lot-to-lot (<4 % variation); Ham’s F-12 showed lower K and Ca but introduced Fe at 0.12 ppm.
• Trace analysis revealed Zn in one DMEM brand (≈22 ppb) and elevated Co, Zn and Sr in Ham’s F-12, reflecting medium fortification strategies.
• IntelliQuant semiquantitative scans provided an elemental heat-map overview, facilitating rapid screening for unexpected contaminants.

Benefits and Practical Applications

The 7900 ICP-MS method delivers robust, sensitive and reproducible multi-element analysis of CDM. Its wide dynamic range allows concurrent measurement of major and trace elements in a single run. High throughput, minimal operator setup and strong interference removal support routine quality control, batch release testing and media formulation optimization.

Future Trends and Potential Applications

• Developing statistical reference profiles for CDM to establish QC benchmarks and acceptance criteria.
• Applying ICP-MS workflows to novel cell and gene therapy media with expanded elemental panels.
• Integrating real-time IntelliQuant monitoring for in-process control in bioreactor systems.
• Expanding automation and data analytics for high-throughput screening of media additives and trace impurity sources.

Conclusion

This study demonstrates that the Agilent 7900 ICP-MS with ORS4 He-KED mode offers a reliable, high-performance solution for the multi-element analysis of chemically defined cell culture media. The method’s accuracy, stability and ease of use make it well suited for media development, optimization, and quality surveillance in biopharmaceutical manufacturing.

References

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