Quantitative analysis of morphine in dried blood spots by using morphine-d3 pre-impregnated dried blood spot cards

• Pre-impregnation by immersing the DBS card in an IS-solution was investigated.
• We demonstrated equal results when using IS impregnated or non-impregnated cards.
• We demonstrated that the impregnated IS homogeneity is acceptable (CV 5%).
• We demonstrated that the IS homogeneity is not disturbed by blood spotting.

Two different internal standard dried blood spot (DBS) pre-impregnation procedures (prior to blood spotting) were investigated. In the first procedure DBS pre-impregnation is performed by immersing the DBS card fully into an internal standard solution. In the second procedure pre-impregnation is performed by pipetting a certain volume of an internal standard solution onto the DBS card. Morphine-d3 was used as the model compound for all experiments. The pre-impregnation procedure by immersing was further investigated with respect to homogeneity of impregnation, influence of different blood spotting techniques and the influence of spotting different blood volumes on the internal standard distribution, calibration and stability of pre-impregnated cards. Finally, the immersing procedure was used for the analysis of morphine in dried blood spots and the results were compared to the conventional procedure in which the internal standard morphine-d3 was added to the extraction solvent. The new pre-impregnated cards couple simplicity of operation and convenient use in the field to results equivalent to the conventional procedure.

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