Unit:
Κατεύθυνση Χημική Ανάλυση-Έλεγχος ΠοιότηταςLibrary of the School of Science
Supervisors info:
Μαρκοπούλου Αικατερίνη, Αναπληρώτρια Καθηγήτρια, Εργαστήριο Φαρμακευτικής Ανάλυσης, Τμήμα Φαρμακευτικής, Αριστοτέλειο Πανεπιστήμιο Θεσσαλονίκης
Original Title:
Ανάπτυξη μεθόδου προσδιορισμού υδροκορτιζόνης σε πλάσμα αίματος με εφαρμογή υγρής χρωματογραφίας συζευγμένης με φασματομετρία μάζας
Translated title:
Development of a method of determination of hydrocortisone in blood plasma using liquid chromatography - mass spectrometry
Summary:
A rapid, reliable and sensitive liquid chromatography-mass spectrometry (LC-MS) method was developed for the quantification of free cortisol in blood plasma.
Prednisolone acetate was used as internal standard. The purification of the samples was performed by solid phase extraction (SPE) technique. The extract was analyzed by an electrospray source and a quadrupole analyzer (ESI-Q) using the negative ion addition method (m/z=397 for cortisol and m/z=437 for prednisolone acetate). Chromatographic conditions included a reversed-phase column (C18) and isocratic elution (0.5mL/min flow, ACN:H2O 50:50 v/v). For the development of the method, three series of standard solutions were prepared: A. pure standards, B. standards submitted to SPE and C. standards prepared by blood serum spiking, and after measuring in LC/MS, their corresponding reference curves were constructed by the method of least squares.
Cortisol recoveries were 91.9-111.5% and %RSDs were 6.0-15.2%, respectively. The detection limit for cortisol was 0.85ng/mL and the quantification limit was 2.81ng/mL. The analytical method showed good linearity from 10.8 to 345.6ng/mL. The correlation coefficient (r2) was greater than 0.9981.
Main subject category:
Science
Keywords:
cortisol, prednisolone acetate, blood plasma, SPE, LC-MS
