Carbon dots (CDs) are emerging as
efficient optical probes. However,
their application potential for clinical diagnosis has not been adequately
explored. Herein, we examined the suitability of pyroglutamate CDs
for detecting glucose, cholesterol, and alcohol in blood serum through
their peroxidative activity in the respective enzyme-catalyzed reactions
following fluorometric and colorimetric approaches. In buffer, the
CD’s fluorescence intensity (λex 354nm) enhanced over 115% after interaction with the enzyme proteins due
to different lifetime components on its surface. The enhancement was
also linked to FRET with the proteins (λex 274nm for TRP/TYR). The electrostatic interactions, as revealed from the
zeta potential study, generated binding energy (ΔG, kcal/mol) in the range of −5.8 to −6.3 and greatly
shifted the protein’s secondary structure to β-strand
contents. The CD’s fluorescence in the blood serum medium was
also enhanced where serum’s particulate components contributed
to the emission. All these subvert fluorescence emissions could be
substantially cleaned for detection of peroxide generated in the enzymatic
reaction by filtering the serum particulates and redox proteins prior
to the addition of CDs to the reaction systems. The CD, however, could
complement well in ABTS-based (absorbance at λmax 414nm) colorimetric reaction in blood serum without introducing protein
or particle separation steps for sensitive detection of peroxide.
The limit of detection, dynamic range, and sensitivity discerned for
peroxide in the glucose oxidase-catalyzed reaction system were 183
μM, 0.02–0.10 mM (R
2 = 0.98),
and 0.2482 AU mM–1, respectively. Overall, these
findings will guide clinical application of the peroxidatic CDs to
detect various analytes in blood serum following fluorometric- and
colorimetric-based principles.