Luciane Maria Ribeiro Neto

Experiência de uma instituição de ensino superior no projeto de coleta de medicamentos em desuso

The university extension program involves a process connecting teaching and research to the society. Aiming to integrate university pharmacy with the “5Rs University Extension Program”, it was collected unused medicines from the academic community. The collected material was evaluated quali- and quantitatively from September/2013 to February/2015. As a result, a high amount of synthetic allopathic medicine was 97.3% of the total sampled, being 43% as non-prescribed medicine and the pharmacological classes mostly discharged were the non-steroidal anti-inflammatory (20.4%), followed by the antimicrobials (16.7%). The critical analysis (SWOT) allowed identifying the strong points, opportunities, weaknesses and threats to the project, as well as a proposal to implant improvements. This activity allowed the pharmacy students to develop a critical census about the rational use of medicine (RUM) and identify the role of the pharmaceutical in the promotion of URM and the proper disposal of medicines.

Determination of free urinary cortisol in cushing's syndrome using reversed-phase high performance liquid chromatography

Determination of free urinary cortisol is a test of choice in the diagnosis of Cushings syndrome. In this study, cortisol was quantified using reversed-phase high-performance liquid chromatography (RP-HPLC) in urine samples previously extracted with ether and using triamcinolone acetonide as internal standard (IS). A BDS-Hypersil-C18® column, water-acetonitrile (72:28; v/v), with a flow rate of 1.0 mL/min and detection at 243 nm were used. This method showed to be both effective and efficient, with sensitivity and linearity ranging from 2.50 to 150 μg/L, and can be used in substitution to the radioimmunoassay technique within this concentration range.

Emprego da cromatografia líquida de alta eficiência na determinação de cortisol sérico em substituição à técnica de radioimunoensaio

BACKGROUND: The quantification of cortisol in different organic fluids has not only been applied to different human nosological conditions as a diagnostic aid but it has also been used in clinical research. In clinical application, cortisol is routinely measured by radioimmunoassay (RIA). In the determination of free urinary cortisol this technique has been replaced by the high-performance liquid chromatography mainly in the diagnosis of Cushing syndrome. As to serum cortisol determination, there is no evidence of the application of liquid chromatography as a substitute for other analytical techniques. OBJECTIVE: The development of an analytical methodology using reversed-phase high-performance liquid chromatography (RP-HPLC) to determine serum cortisol levels as a substitute for RIA in order to reduce radioactive waste. MATERIAL AND METHODS: Cortisol was directly quantified by RP-HPLC in previously ether-extracted serum samples. Triamcinolone acetonide was used as internal standard (IS). The chromatographic separation was developed in a BDS-Hypersil-C18® column (125 x 4 mm, 5 µm) using water-acetonitrile (72:28; v/v) as mobile phase at 1 ml/min and steroid peaks were measured at 243 nm. RESULTS: Cortisol and IS presented retention time of 3.4 and 7.1 min, respectively. The precision was less than 10% and accuracy was less than 4%. DISCUSSION: The method was effective and efficient, with good sensitivity and linearity in the concentration range of 2.5 to 60.0 µµg/dl. CONCLUSION: The present methodology substitutes RIA at clinical application.

Aplicação da cromatografia a líquido em substituição à técnica de radioimunoensaio como auxílio diagnóstico visando ao gerenciamento de resíduos de serviço da saúde em laboratório de pesquisa

Designing a Health Care Service Waste Management Plan, according to the RDC 306 rules, is a responsibility of all those who produce such waste. Since radioimmunoassay (RIA) is one of the most employed techniques, we studied the impact of replacing this technique by liquid chromatography (HPLC) with regard to the reduction of the radioactive residues routinely produced by the Unifesp steroid laboratory. The residues produced by the determination of serum cortisol and 17 α-hydroxyprogesterone were classified, and those belonging to groups B and C were evaluated. We observed that, when RIA is used, chemical residues (group B) and radioactive waste (group C) are produced, whereas HPLC generates only chemical residues. Adequation of these techniques showed to be advantageous, by significantly reducing the time of analysis and mainly by eliminating and/or reducing the generation of radioactive waste, encouraging its application to other methodologies, as well as its adoption by other research units.

Aplicação da cromatografia líquida de alta eficiência para quantificação direta da 17

A determinacao dos niveis sericos de 17±-hidroxiprogesterona (17OHP) e essencial no diagnostico laboratorial da hiperplasia adrenal congenita (CAH) por deficiencia de 21-hidroxilase. Essa condicao e responsavel por pseudo-hermafroditismo em meninas e precocidade sexual em ambos os sexos. A 17OHP foi quantificada diretamente empregando-se cromatografia liquida de alta eficiencia no modo fase reversa (RP-HPLC) em amostras de soro previamente extraidas com eter. Utilizou-se coluna ODS-Hypersil® e fase movel composta de agua-metanol (4:6 v/v) com vazao de 1,0 mL/min. A determinacao deu-se em 246 nm. A 17OHP apresentou tempo de retencao de 5,6 min. O metodo mostrou-se eficaz e eficiente com sensibilidade e linearidade (r2 = 0,9993) na faixa de concentracao estudada de 500 a 100.000 ng/dL. As precisoes intra e interensaios foram respectivamente 4,4 e 8,1%. O metodo pode ser empregado na rotina laboratorial da avaliacao desse esteroide para diagnostico de CAH, sendo necessario o emprego do radioimunoensaio(RIA) apenas para quantificar amostras com valores abaixo de 500 ng/dL.