In this study, hair externally contaminated with fentanyl underwent decontamination with combinations of parameters based on a 24 fractional factorial block design DoE matrix.
Optimal methods for hair analysis are often debated. Previous work in this laboratory demonstrated that the statistical technique known as Design of Experiments (DoE) is useful for such optimization. DoE evaluates both the individual roles and the combinatorial associations among multiple independent variables (i.e., hair pretreatment parameters) and a dependent variable (i.e., drug recovery from hair). In the current study, The parameters of interest included aqueous wash solvent (1% sodium dodecyl sulfate (SDS) or water), organic wash solvent (dichloromethane or methanol), number of consecutive washes (one or three), sequence of washes (aqueous first or organic first) and wash time (30 s or 30 min). The optimal method for decontaminating fentanyl from the hair surface was found to be one 30-min wash with dichloromethane followed by one 30-min wash with water. Pretreatment parameters were optimized with a 23 full factorial DoE matrix using authentic hair reference material (HRM), which consisted of pooled drug user hair diluted to a known concentration of fentanyl with drug-free hair. The factors of interest were extraction solvent/sample weight ratio (12.5 or 25 µL/mg), hair particle size (pulverized or 1 mm segments) and extraction time (2 or 24 h). The most effective pretreatment method for fentanyl consisted of pulverizing the hair prior to a 2-h extraction in a 25 µL/mg extraction solvent volume/sample weight ratio. Finally, using the optimized pretreatment methods, fentanyl containing authentic HRM was extracted using aqueous base, solvent and enzymatic hair extraction methods, where it was determined that the aqueous base technique was most effective for recovery of fentanyl. These experiments further demonstrate the value of DoE and authentic HRM in method development for forensic hair analysis. (Publisher abstract provided)
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