Non-invasive transdermal sampling and analysis device incorporating redox cofactors

A method of determining levels of an analyte in a biological fluid sample, the method comprising:

• providing at least one disruptor mounted on a substrate, wherein the at least one disruptor is configured to generate a localized heat capable of altering permeability characteristics of a stratum corneum layer of skin of an organism;
• providing a reservoir configured to collect and contain the biological fluid sample;
• providing a sensing element, comprising:
• at least two sensing electrodes, wherein a surface of at least one of the sensing electrodes is coated with a sensing layer comprising an enzyme immobilized within a hydrogel;
• and
• applying at least one non-bound layer of a cofactor atop the sensing layer, wherein the cofactor is configured to become free-floating upon introduction of the biological fluid sample to the sensing element and catalyzes a reaction to determine levels of an analyte in the biological fluid sample;
• and
• providing the biological fluid sample to the sensing element and the at least one non-bound layer of the cofactor.