Silvia Bérgamo Vázquez (Presenter)
University Hospital of South Manchester
Bio: I graduated in Pharmacy from the University of Santiago de Compostela, Spain, in 2012. In 2013 I started as a Clinical Biochemistry resident in Vall d’Hebrón Hospital. My role includes different analytical method development as well as providing advice to clinicians, authorisations of patient results and overseeing a section of the laboratory. I usually do night shifts in the Emergency laboratory. I am especially interested in working with chromatographic and mass spectrometry techniques so I have completed a trainee period in University Hospital of South Manchester over three months with Dr. Brian Keevil. At the moment, I am working as a Clinical Biochemistry resident in Vall d’Hebrón Hospital in Barcelona.
Authorship: Silvia Bérgamo Vázquez, Brian Keevil
University Hospital of South Manchester
| Short Abstract Evidence suggests that Primary hyperaldosteronism subtype classification through specific urinary steroid quantification may help guide treatment. This work describes the development and validation of a LC-MS/MS method to quantify urinary 18-hydroxycortisol,18-oxocortisol and tetrahydroxyaldosterone. Urine samples were prepared using liquid- liquid extraction. Chromatographic separation was achieved over 4.5 min using a phenyl 1.7 µm 2.1x 100 mm column and detected using a Waters TQMS. No interference or ion suppression were observed. Intra- and inter-assay imprecision was within the admissible range. Linearity and the LOQs covered clinically relevant concentrations. This method has been successfully developed and validated for clinical analysis. |
Long Abstract
Background:
Primary hyperaldosteronism (PA) is the most common form of secondary hypertension. First of all, the most important step is to establish the correct subtype diagnosis, since the treatment will vary accordingly.
In unilateral hyperproduction cases , surgical treatment will be an option, whereas for bilateral hyperproduction , patients could receive medical treatment with a mineralocorticoid receptor antagonist.
The most reliable method to identify the principal subtypes of PA is adrenal vein sampling. Unfortunately this technique requires specialist review and high-cost equipment, so this test is not available in all hospitals.
Evidence suggests that PA subtype classification through specific urinary steroid quantification may help guide treatment. This work describes developed and validated a LC-MS/MS method to quantify urinary 18-hydroxycortisol (18-OHF),18-oxocortisol (18-OXO) and tetrahydroxyaldosterone (TH-ALDO).
Methods:
To 150 µL of standard/QC/sample, deuterated internal standards, E-coli (10 µL) and buffer (150 µL) were added. After incubation samples were extracted with methyl tert-butyl ether (MTBE), the supernatant dried and reconstituted in 30% methanol. Chromatographic separation was achieved over 4.5 min using a phenyl 1.7 µm 2.1x 100 mm column and detected using a Waters TQMS.
Results:
No interference or ion suppression were observed. Intra- and inter-assay imprecision (was within the admissible ranged) from 3.2 – 10.9% over 100-600 µg/L (18-OHF), 2.5-15.0 µg/L (18-OXO) and 50-300 µg/L (TH-ALDO). Linearity and the LOQs covered all clinically relevant concentrations.
Conclusions:
This method has been successfully developed and validated for use in routine clinical biochemistry laboratories.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | no | |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | no |