Background and importance We have previously quantified and identified the content of doxorubicin both as a pharmaceutical and laboratory substance dissolved in complemented fluorobrite (chromophorous food media). In the present study we attempted to measure the same preparations in the efluate of cultured cell lines using the DrugLog device (Pharmacolog AB Sweden) as part of our quality assurance process.
Aim and objectives Demonstration of the suitability of this method to determine the concentration of doxorubicin in the above-specified preparation in a qualitative and quantitative manner.
Material and methods Triple-negative breast cancer cell lines MDA-MB-231 were cultured with a density of 1×105 cells/mL in Dulbecco’s Modified Eagle Medium with 10% fetal bovine serum. Doxorubicin-complemented fluorobrite was used as culture media. After quantitative determination of the fluorobrite concentration with the Druglog, doxorubicin (as a second chromophore in our solutions) was added. For this purpose either Adrimedac (liquid form of doxorubicin, Doxorubicin Aurobindo (powder form) or Doxorubicin Sigma Aldrich (laboratory substance in powder form) in complemented fluorobrite was used. After calibration, concentration of the respective solutions was measured.
Results The three different preparations have shown the following results:
Adrimedac: relative deviation after day 1: 3.3% (n=18), day 2: 16.46% (18);
Doxorubicin Aurobindo: day 1: 1.84% (18); day 2: 8.89% (18);
Doxorubicin Sigma Aldrich: day 1: 22.4% (18); day 2: 32.6% (18).
According to our observation, the deviations were surprisingly large and the drug concentrations in all solutions were lower than expected, compared to the concentrations of doxorubicin in the preparations without cells. The preparations with the laboratory substance were the most rapidly degraded.
Conclusion and relevance From our results it can be concluded that the Druglog is suitable to provide reproducible quantitative measurements of doxorubicin concentrations from the efluate of cell preparations without further filtration of the solutions. In future experiments we will compare these results using a different method, in order to explain the difference in the deviations of doxorubicin in the solutions between the pharmaceutical and the laboratory grade of doxorubicin. This method can be used for further research of cytostatics preparations in hospital pharmacies.
Conflict of interest No conflict of interest
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