The plasma protein binding of the endogenous glucocorticosteroids is of vital importance for the concentrations in hair and saliva

Abstract

BackgroundThe endogenous glucocorticosteroid cortisol (F) and its metabolite cortisone (E) are known to be involved in stress adaption and anti-inflammatory and immune regulatory effects. The ratios of F to E in the matrices serum, hair and saliva are different. The shift of this ratio by the enzyme activity of 11β-hydroxysteroid-dehydrogenase, which inactivates cortisol, was often discussed. The aim of our study was to calculate the contribution of the plasma protein binding (PPB) to this shift. The PPB of F is known to be 96% of the total F-Concentration in serum. The PPB of E was not analyzed in previous studies. MethodsOur study was designed to evaluate the correlation of corticosteroid concentrations in serum (total and free), hair and saliva. The samples were self-collected by the author (A.K.) monthly over a pregnancy cycle (1st samples before pregnancy, 8 samples during pregnancy and 5 samples postpartum). Serum protein binding was calculated from the determination of the total hormone concentrations of F and E (protein bound and unbound) and the free hormone concentrations in serum. The samples were processed by ether extraction and ultrafiltration. Hair samples were extracted with methanol and purified by solid-phase extraction. Saliva samples were collected using Salivette® collection system. The concentrations of F and E were measured by liquid chromatography-mass spectrometry with LODs for free serum, total serum, hair and saliva of F: 0.11ng/mL, 2.13ng/mL, 1.6pg/mg, 0.08ng/mL and E: 0.12ng/mL, 0.54ng/mL, 2.1pg/mg, 0.09ng/mL, respectively. Results and discussionThe serum concentrations (free and total) of both glucocorticosteroids rise up continuously during the time of pregnancy and decrease after delivery. The free and total serum concentrations were well correlated. No change was detected for the intensity of PPB of F. In contrast, the PPB of E decreases from 86.3% to 80.7% during pregnancy. The concentration ratios of F to E change from 3:1 in total serum to 1:1 in free serum. For hair samples, an increase of F and E in proximal segments was confirmed with the highest concentration 6.5weeks postpartum. Independently, corticosteroid concentrations in corresponding hair segments were found to be reduced with increasing distance from the root; an average decline of F and E by half in 5 and 6months was estimated, respectively. The counter effect of the mechanisms incorporation and wash-out is clearly visible. For saliva samples a good correlation with free, non-protein bound serum concentration was detected.