Plasma eicosanoid profiles determined by high-performance liquid chromatography coupled with tandem mass spectrometry in stimulated peripheral blood from healthy individuals and sickle cell anemia patients in treatment

Author(s)

Galvao, Alyne Favero ^[1] ; Petta, Tania ^[1] ; Flamand, Nicolas ^[2] ; Bollela, Valdes Roberto ^[3] ; Silva, Celio Lopes ^[4] ; Jarduli, Luciana Ribeiro ^[1] ; Ribeiro Malmegrim, Kelen Cristina ^{[1, 5]} ; Simoes, Belinda Pinto ^[3] ; Beraldo de Moraes, Luiz Alberto ^[6] ; Faccioli, Lucia Helena ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Dept Anal Clin Toxicol & Bromatol, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil

^[2] Univ Laval, Fac Med, Dept Med, Ctr Rech, Inst Univ Cardiol & Pneumol Quebec, 2725 Chemin St Foy, Ste Foy, PQ G1V 4G5 - Canada

^[3] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Clin Med, Av Bandeirantes 3900, BR-14049900 Ribeirao Preto, SP - Brazil

^[4] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, Av Bandeirantes 3900, BR-14049900 Ribeirao Preto, SP - Brazil

^[5] Univ Sao Paulo, Fac Med Ribeirao Preto, Ctr Reg Hemoterapia Hosp Clin, Ctr Terapia Celular, Rua Tenente Catao Roxo 2501, BR-14049900 Ribeirao Preto, SP - Brazil

^[6] Univ Sao Paulo, Dept Quim, Fac Filosofia Ciencias & Letras Ribeirao Preto, Av Bandeirantes 3900, BR-14049901 Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: ANALYTICAL AND BIOANALYTICAL CHEMISTRY; v. 408, n. 13, p. 3613-3623, MAY 2016.
• Web of Science Citations: 10

Abstract

Eicosanoids play an important role in homeostasis and in the pathogenesis of various human diseases. Pharmacological agents such as Ca2+ ionophores and Ca2+-ATPase inhibitors, as well as natural agonists such as formylmethionine-leucyl-phenylalanine (fMLP), can stimulate eicosanoid biosynthesis. The aims of this work were to develop a method to determine the eicosanoid profile of human plasma samples after whole blood stimulation and to assess differences between healthy and sick individuals. For this purpose, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was partially validated for the quantification of 22 eicosanoids using human plasma from healthy volunteers. In addition, we optimized a method for the stimulation of eicosanoids in human whole blood. LC-MS/MS analyses were performed by negative electrospray ionization and multiple reaction monitoring. An assumption of linearity resulted in a regression coefficient a parts per thousand yen0.98 for all eicosanoids tested. The mean intra-assay and inter-assay accuracy and precision values had relative standard deviations and relative errors of a parts per thousand currency sign15 %, except for the lower limit of quantification, where these values were a parts per thousand currency sign20 %. For whole blood stimulation, four stimuli (fMLP, ionomycin, A23187, and thapsigargin) were tested. Results of the statistical analysis showed that A23187 and thapsigargin were potent stimuli for the production or liberation of eicosanoids. We next compared the eicosanoid profiles of stimulated whole blood samples of healthy volunteers to those of patients with sickle cell anemia (SCA) under treatment with hydroxyurea (HU) or after chronic red blood cell (RBC) transfusion. The results indicate that the method was sufficient to find a difference between lipid mediators released in whole blood of SCA patients and those of healthy subjects, mainly for 5-HETE, 12-HETE, LTB4, LTE4, TXB2, and PGE(2). In conclusion, our analytical method can detect significant changes in eicosanoid profiles in stimulated whole blood, which will contribute to establishing the eicosanoid profiles associated with different inflammatory and infectious diseases. (AU)

• FAPESP's process: 14/07125-6 - New functional aspects of eicosanoids
• Grantee: Lúcia Helena Faccioli
• Support Opportunities: Research Projects - Thematic Grants