Simulation of the preparation of an injectable anticancer medicine using a Media Fill Test coupled with Fluorescein: a technical validation method of preparation quality

Aim

To assess risks of microbiological and chemical contamination (Media Fill Test and fluorescence detection) and validate the quality of the preparation process (quantitative dosage of fluorescein in the final preparation).

Method

Simulation preparation of an injectable anticancer replacing raw materials with a fluorescein solution (active substance) and a culture medium (solvent) in worst case conditions.

• Manipulations : each manipulator realizes 9 different preparations by diluting of different fixed concentrations of fluorescein in the culture medium
• Contamination evaluation : 
  • chemical : revelation of fluorescence with a UV lamp
  • microbiological : incubation of preparations (after fertility test)
• Quality assessment of the various diluting processes : quantitative dosage of the preparations by UV Spectrophotometry (validation of the method in the culture medium)
• Validation of the manipulators on 4 criteria: manipulation technique (by a trainer), revelations of fluorescence outcomes, dosing of fluorescein and media fill test

Results 

Validation of the assay method: 3 days, 3QC, CV¡Ü5%, y = 0.2153x + 0.0016, R2 = 0.9997
10 manipulators : 4 pharmacists, 6 technicians
Traces of fluorescein: 12 (fields, gloves, sites of injection, compresses)
Dosages of fluorescein: average derived variance compared with the expected theoretical concentration : 4.95%, (technicians: 3.85% ; pharmacists: 8.8%)
Media fill test: 100% negatives results
An technician and 3 pharmacists are not validated for a variance on a dosage > 10 %.

Conclusion
 
This simulation is a complete tool for training and manipulators validation which also enable to review our laboratory practices. It allowed us to work out the imprecision associated with the use of some too large volume of syringe and the mixing time required to obtain homogeneous preparations.