Abstract
The aim of this study was to develop the analytical method of the isolation and determination of cortisol from the saliva of depressed women. The material for introduct study was obtained from 10 healthy subjects and was collected in the evening hours, about 8 p.m. when the concentration of cortisol is the lowest. The extraction of the cortisol from saliva was made by liquid-liquid extraction (LLE) by dichloromethane. The level of the hormone was estimated by HPLC with UV detection at 240 nm. The mobile phase contained acetonitrile and water (35:65; v/v) at a flow of 1 mL min-1. The time of retention for cortisol was 2.9 min and for carbamazepine (internal standard) was 4.9 min. The estimated analytical method was validated. Linearity of the calibration curve (r = 0.9997) was obtained in the concentration range of 5–200 ng mL-1 of cortisol in saliva and the limit of detection (S/N = 3) was 1 ng mL-1. The RSD for the intra-assay study varied between 1.1 and 9.9% and in the inter-assay study did not exceed 6.8%. In the intra-assay and inter-assay of the study the accuracy was about 100. The efficiency of the method was made for two different concentrations of cortisol 15 and 125 ng ml-1. The precision for the lower concentration was 7.1% and 8.2% adequate for cortisol spiked after and before the extraction. For the concentration 125 ng ml-1 the efficiency was 3.4% and 2.6%. The estimated method of extraction was applied to analysis of saliva samples without interference peaks. It gave satisfying results and may be a useful toole for monitoring the changes in salivary cortisol.
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