Detection of Porphyrins in Hair Using Capillary Liquid Chromatography-Mass Spectrometry

Int J Mol Sci. 2022 Jun 2;23(11):6230. doi: 10.3390/ijms23116230.

Abstract

Unlike humans, some animals have evolved a physiological ability to deposit porphyrins, which are pigments produced during heme synthesis in cells, in the skin and associated integument such as hair. Given the inert nature and easiness of collection of hair, animals that present porphyrin-based pigmentation constitute unique models for porphyrin analysis in biological samples. Here we present the development of a simple, rapid, and efficient analytical method for four natural porphyrins (uroporphyrin I, coproporphyrin I, coproporphyrin III and protoporphyrin IX) in the Southern flying squirrel Glaucomys volans, a mammal with hair that fluoresces and that we suspected has porphyrin-based pigmentation. The method is based on capillary liquid chromatography-mass spectrometry (CLC-MS), after an extraction procedure with formic acid and acetonitrile. The resulting limits of detection (LOD) and quantification (LOQ) were 0.006-0.199 and 0.021-0.665 µg mL-1, respectively. This approach enabled us to quantify porphyrins in flying squirrel hairs at concentrations of 3.6-353.2 µg g-1 with 86.4-98.6% extraction yields. This method provides higher simplicity, precision, selectivity, and sensitivity than other methods used to date, presenting the potential to become the standard technique for porphyrin analysis.

Keywords: CLC-MS; animal pigmentation; fluorescence; flying squirrels; hair; mammals; natural porphyrins.

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Liquid
  • Hair / chemistry
  • Mammals
  • Mass Spectrometry
  • Porphyrins* / chemistry

Substances

  • Porphyrins

Grants and funding

The Spanish Ministry of Science and Innovation, JJCC Castilla—La Mancha and University of Castilla—La Mancha are gratefully acknowledged for funding this work with Grants PID2019-104381GB-I00, JCCM SBPLY/17/180501/000262 and 2020-GRIN-28882, respectively.