Paper-Based Device for Rapid Visualization of NADH Based on Dissolution of Gold Nanoparticles

ACS Appl Mater Interfaces. 2015 Jul 15;7(27):15023-30. doi: 10.1021/acsami.5b04104. Epub 2015 Jul 2.


We describe a paper-based device that enables rapid and sensitive room-temperature detection of dihydronicotinamide adenine dinucleotide (NADH) via a colorimetric readout and demonstrate its value for monitoring NAD+-driven enzymatic reactions. Our system is based on NADH-mediated inhibition of gold nanoparticle (AuNPs) dissolution in a Au3+-cetyltrimethylammonium bromide (CTAB) solution. We fabricated a device consisting of a mixed cellulose ester paper featuring a wax-encircled, AuNP-coated film atop a cotton absorbent layer sandwiched between two plastic cover layers. In the absence of NADH, the Au3+-CTAB complex dissolves the AuNP layer completely, generating a white color in the test zone. In the presence of NADH, Au3+ is rapidly reduced to Au+, greatly decreasing the dissolution of AuNPs and yielding a red color that becomes stronger at increasing concentrations of NADH. This device exploits capillary force-assisted vertical diffusion, allowing us to apply a 25 μL sample to a surface-confined test zone to achieve a detection limit of 12.5 μM NADH. We used the enzyme glucose dehydrogenase as a model to demonstrate that our paper-based device can monitor NAD+-driven biochemical processes with and without selective dehydrogenase inhibitors by naked-eye observation within 4 min at room temperature in a small sample volume. We believe that our paper-based device could offer a valuable and low-cost analytical tool for monitoring NAD+-associated enzymatic reactions and screening for dehydrogenase inhibitors in a variety of testing contexts.

Keywords: NAD+-driven enzymatic reactions; colorimetric screening; dehydrogenase inhibitors; dihydronicotinamide adenine dinucleotide; dissolution of gold nanoparticles; paper-based device.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Colorimetry / instrumentation*
  • Colorimetry / methods
  • Equipment Design
  • Equipment Failure Analysis
  • Gold / chemistry*
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • NAD / analysis*
  • NAD / chemistry
  • Paper*
  • Reagent Strips*
  • Reproducibility of Results
  • Sensitivity and Specificity


  • Reagent Strips
  • NAD
  • Gold