Pharmaceutical Research

, 28:2920 | Cite as

Time-Correlated Single Photon Counting For Simultaneous Monitoring Of Zinc Oxide Nanoparticles And NAD(P)H In Intact And Barrier-Disrupted Volunteer Skin

  • Lynlee L. Lin
  • Jeffrey E. Grice
  • Margaret K. Butler
  • Andrei V. Zvyagin
  • Wolfgang Becker
  • Thomas A. Robertson
  • H. Peter Soyer
  • Michael S. Roberts
  • Tarl W. ProwEmail author
Research Paper



There is a lack of relevant, non-animal alternatives for assessing exposure and toxicity of nanoparticle-containing cosmetics, e.g. sunscreens. Our goal was to evaluate timecorrelated single photon counting (TCSPC) for simultaneous monitoring of zinc oxide nanoparticles (ZnO-NP) and the metabolic state of volunteer skin.


We separated the fluorescence lifetime signatures of endogenous fluorophore signals (i.e. nicotinamide adenine dinucleotide phosphate, NAD(P)H and keratin) and the ZnO-NP signal using advanced TCSPC to simultaneously determine ZnO-NP penetration profiles and NAD(P)H changes in subjects with altered barrier function, including tape-stripped skin and in psoriasis or atopic dermatitis lesions.


We detected no ZnO-NP penetration into viable human skin in any group. ZnO-NP signal was significantly increased (p < 0.01) on the surface of tape-stripped and lesional skin after 4 and 2 h of treatment, respectively. Free NAD(P)H signal significantly increased in tape-stripped viable epidermis treated for 4 h of ZnO-NP compared to vehicle control. No significant NAD(P)H changes were noted in the lesional study.


TCSPC techniques enabled simultaneous, real-time quantification of ZnO-NP concentration and NAD(P)H via non-invasive imaging in the stratum corneum and viable epidermis of volunteers.


human skin metabolism multiphoton microscopy sunscreen zinc oxide nanoparticle 



atomic absorption spectroscopy


arbitrary unit


caprylic/capric triglycerides


fluorescence lifetime imaging microscopy


inductively coupled plasma-optical emission spectroscopy


instrument response function


potassium di-hydrogen phosphate


multiphoton-excited photoluminescence


multiphoton tomography


multiphoton tomography with fluorescence lifetime imaging microscopy


nicotinamide adenine dinucleotide phosphate


phosphate-buffered saline


second harmonic generation


time-correlated single photon counting


transmission electron microscope


transepidermal Water Loss




zinc oxide nanoparticles



We would like to thank the National Health and Medical Research Council of Australia (ID# 569694) and the United States Air Force Asian Office of Aerospace Research and Development for funding. We also thank Corinne Yoong for recruiting volunteers for the lesion studies.


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Lynlee L. Lin
    • 1
    • 2
  • Jeffrey E. Grice
    • 1
  • Margaret K. Butler
    • 3
  • Andrei V. Zvyagin
    • 4
    • 5
  • Wolfgang Becker
    • 6
  • Thomas A. Robertson
    • 7
  • H. Peter Soyer
    • 2
  • Michael S. Roberts
    • 1
    • 7
  • Tarl W. Prow
    • 1
    • 2
    • 8
    Email author
  1. 1.Therapeutics Research Centre, School of Medicine Princess Alexandra HospitalUniversity of QueenslandBrisbaneAustralia
  2. 2.Dermatology Research Centre, School of Medicine Princess Alexandra HospitalUniversity of QueenslandBrisbaneAustralia
  3. 3.Australian Institute for Bioengineering & NanotechnologyUniversity of QueenslandBrisbaneAustralia
  4. 4.Department of Physics, Centre of MQ PhotonicsMacquarie UniversitySydneyAustralia
  5. 5.Centre for Biophotonics and Laser Science School of Physical SciencesUniversity of QueenslandBrisbaneAustralia
  6. 6.Becker & Hickl GmbHBerlinGermany
  7. 7.Therapeutics Research Centre School of Pharmacy & Biomedical SciencesUniversity of South AustraliaAdelaideAustralia
  8. 8.Therapeutics Research & Dermatology Research Centres School of Medicine, Princess Alexandra HospitalUniversity of QueenslandWoolloongabbaAustralia

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