Rapid saliva processing techniques for near real-time analysis of salivary steroids and protein

J Clin Lab Anal. 2008;22(6):395-402. doi: 10.1002/jcla.20281.

Abstract

Introduction: Point-of-care (POC) measurements using saliva samples have immense potential to assess systemic health and wellbeing, but sample viscosity and contaminants can affect analyses. We sought a portable clean-up method for whole saliva appropriate for use with POC measurement techniques such as biosensors.

Methods: Whole saliva from each of 13 male subjects was split into 5 fractions. Each fraction was treated with a different clean-up process: a freeze-thaw-centrifuge (FTC) step; centrifugation alone; or passage through a Mini-UniPrep polyethersulfone filter, cotton Salivette, or foam Oracol device. Following clean-up, each subject's treated saliva fractions were assayed for cortisol, testosterone, dehydroepiandrosterone (DHEA), and protein concentrations. The effects of clean-up methods on nonspecific binding (NSB) in a biosensor were also assessed.

Results: Compared with FTC, no analytes were affected by centrifugation alone. Cotton Salivettes significantly altered all analytes, with increases in cortisol (+64%), testosterone (+126%), and DHEA (off-scale) levels, and decreased protein (-21%) and biosensor NSB (-75%). Oracol foam devices decreased DHEA levels by 28%. Mini-UniPrep filtration decreased testosterone (-45%) and DHEA (-66%) concentrations while increasing cortisol (+40%).

Conclusion: No method was optimal for all analytes, highlighting the need for validation of saliva treatment methods before their adoption in rapid POC analyses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Centrifugation
  • Dehydroepiandrosterone / analysis
  • Filtration
  • Freezing
  • Humans
  • Hydrocortisone / analysis
  • Male
  • Middle Aged
  • Point-of-Care Systems
  • Radioimmunoassay
  • Saliva / chemistry*
  • Salivary Proteins and Peptides / analysis
  • Specimen Handling / methods*
  • Surface Plasmon Resonance
  • Testosterone / analysis

Substances

  • Salivary Proteins and Peptides
  • Testosterone
  • Dehydroepiandrosterone
  • Hydrocortisone