On-Capillary Surface-Enhanced Raman Spectroscopy: Determination of Glutathione in Whole Blood Microsamples

Autores de IIS La Fe

• Ángel Sánchez Illana

  Autor

• Mayra Pilar Rubio Martinez

  Autor

• Daniel Cuesta Garcia

  Autor

• Jose David Piñeiro Ramos

  Autor

• Máximo Vento Torres

  Autor

• Guillermo Quintas Soriano

  Autor

• Julia Kuligowski

  Autor

Participantes ajenos a IIS La Fe

• Cantarero, Andres
• de la Guardia, Miguel
• Lendl, Bernhard

Grupos

• Perinatología

Abstract

Oxidative stress monitoring in the neonatal period supports early outcome prediction and treatment. Glutathione (GSH) is the most abundant antioxidant in most cells and tissues, including whole blood, and its usefulness as a biomarker has been known for decades. To date, the available methods for GSH determination require laborious sample processing and the use of sophisticated laboratory equipment. To the best of our knowledge, no tools suitable for point-of-care (POC) sensing have been reported. Surface-enhanced Raman spectroscopy (SERS), performed in a microvolume capillary measurement cell, is proposed in this study as a robust approach for the quantification of GSH in human whole blood samples. The use of a silver colloid allowed a highly selective signal enhancement for GSH providing analytical enhancement factors of 3 to 4 orders of magnitude. A highly accurate determination of GSH in whole blood samples with recoveries ranging from 99 to 107% and relative standard deviations less than or equal to 18% were achieved by signal normalization with the intensity of an isotopically labeled internal standard. GSH concentrations were retrieved within 4 min using small-volume blood samples (2 mu L). The developed procedure was applied to the analysis of blood of 20 healthy adults and 36 newborns, obtaining comparable results between literature and those found by SERS and a reference method. The characteristics of this novel tool are suitable for its implementation in a portable optical sensor device enabling POC testing of oxidative stress levels in newborns.

Keywords

• OXIDATIVE STRESS; BIOLOGICAL SAMPLES; AMINO-ACIDS; OXYGEN SUPPLEMENTATION; OXIDIZED GLUTATHIONE; AQUEOUS-SOLUTIONS; PRETERM INFANTS; SENSING METHOD; SCATTERING; SERS