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Urine Storage Under Refrigeration Preserves the Sample in Chemical ORIGINAL ARTICLE J Bras Patol Med Lab, v. 49, n. 6, p. 415-422, dezembro 2013 Urine storage under refrigeration preserves the sample in chemical, cellularity and bacteriuria analysis of ACS Armazenamento da urina, sob refrigeração, preserva a amostra nas análises químicas, celularidade e bacteriuria no EAS Karen Cristina Barcellos Ribeiro1; Bruno Rotondo Levenhagem Serabion2; Eduardo Lima Nolasco3; Chislene Pereira Vanelli4; Harleson Lopes de Mesquita5; José Otávio do Amaral Corrêa6 ABSTRACT Introduction: The analysis of urine abnormal constituents and sediment (ACS) comprises tests of great diagnostic and prognostic value in clinical practice. When the analysis of ACS cannot be performed within two hours after collection, the sample must be preserved in order to avoid pre-analytical interferences. Refrigeration is the most applied technique due to its cost effectiveness. Moreover, it presents fewer inconveniences when compared to chemical preservation. However, changes in ACS may also occur in samples under refrigeration. Objective: To analyze the influence of refrigeration at 2 to 8ºC on the storage of urine samples within 24 hours.Material and method: A total of 80 urine samples were selected from patients admitted at Universidade Federal de Juiz de Fora (UFJF) university hospital, which were tested for ACS at room temperature and stored under refrigeration for 6, 12 and 24 hours. Results: The results showed that refrigeration proved to be effective when compared to samples kept at room temperature, inasmuch as the physical, chemical, microbial and cellularity features were preserved. Nevertheless, crystalluria was present after a 6- hour storage period. Conclusion: The tests revealed that cooling preserved cellularity and chemical characteristics of urine samples for up to 12 hours. Nonetheless, the precipitation of crystals was evident in this storage method. Thus, the possible consequences of storing urine samples for ACS test under these conditions should be included in the analysis report. Key words: urinalysis; urine storage; crystalluria; pyuria; hematuria Introduction. INTRODUCTION Several systemic clinical conditions that may affect renal function and / or change the composition of urine, namely The routine urine test or urinalysis is a vital laboratory tool hydration status, progression and diagnosis of diabetes, in clinical practice. One of the most important urine analysis test hepatopathy, hemolytic anemia or poisoning, may have higher is the screening of abnormal constituents and sediment (ACS). chances of being diagnosed and/or monitored by ACS analysis(2, The ACS test comprises physical (organoleptic features, volume 4-6). Furthermore, the ACS assessment is vital in detecting renal and and density), chemical and microscopic analysis of the urinary sediment(2, 5, 7). This is an inexpensive and commonly available urogenital system diseases such as glomerulonephritis, nephrotic test, which provides a considerable amount of useful information syndrome, cystitis, pyelonephritis, renal failure, lithiasis and even on the diagnosis of metabolic and genitourinary tract disorders(1, 3). cancer(2, 6). First submission on 19/02/13; last submission on 13/08/13; accepted for publication on 12/09/13; published on 20/12/13 1. Pharmacist at Hospital Universitário Sul Fluminense. 2. Graduate in Pharmacy by Universidade Federal de Juiz de Fora (UFJF). 3. Doctoral student in Pharmaceutical Sciences at the Immunoendocrinology Laboratory- Department of Clinical and Toxicological Analysis – Faculty of Pharmaceutical Sciences- Universidade de São Paulo (USP). 4. Clinical nutritionist at Centro de Tratamento de Doenças Renais and Fundação Imepem; MSc student in Brazilian Health at UFJF. 5. MSc in Clinical Analysis by Universidade Santo Amaro (UNISA); adjunct professor at Faculdade Suprema; doctorate student in Brazilian Health at UFJF. 6. PhD in Pathology by Universidade Federal Fluminense (UFF); adjunct professor and coordinator of the Pharmacy Course at UFJF. 415 Urine storage under refrigeration preserves the sample in chemical, cellularity and bacteriuria analysis of ACS In clinical pathology laboratories, all analysis procedures, analysis. The refrigerated samples were compared with those kept including urine tests, are divided into pre-analytical, analytical at room temperature for the performance of physical, chemical and post-analytical phases(8, 17). The preanalytical phase is the and microscopic ACS analysis. step in which fundamental procedures are required in order to guarantee the quality and service performance. It involves the collection, handling, processing, delivery of the sample to MATERIAL AND METHOD analyzers and storage when it is not processed on arrival at the laboratory. It also accounts for 46% to 70% of laboratory errors, Patient and samples which may lead to inaccurate results, thus hindering clinical The urine samples were obtained from adult patients aged (8, 9, 11, 18) diagnosis and patient’s healthcare . from 18 to 60 years from both genders, who were hospitalized Therefore, urine collection (pre-analytical phase) should at Universidade Federal de Juiz de Fora (UFJF) - university follow basic procedures in order to obtain a sample that reflects hospital and who had ACS test requests. physicochemical, cellular and microbiological changes(2, 7). Ideally, 80 urine samples containing 80 ml each were from clean- the sample should be collected after cleaning the genitourinary catch midstream one-time urine collection after nocturnal tract through spontaneous and preferably long midstream urine concentration. They were stored in containers standardized by the after nocturnal concentration (first morning urination). The World Health Organization (WHO) and immediately dispatched to dispatch and sample analysis should occur within two hours after the urinalysis sector of the Clinical Analysis Laboratory from UFJF collection if kept at room temperature(7, 12). University Hospital and analyzed at arrival. According to the Brazilian Association of Technical Standards After the performance of ACS test, the samples were divided (Associação Brasileira de Normas Técnicas- ABNT)(2), when the and stored at different temperatures (refrigeration at 2 to 8ºC and analysis cannot be performed within this period, preservatives at room temperature). Additionally, they were assessed at different should be used in order to conserve the characteristics of the urine periods (6, 12 and 24 hours). and prevent inaccurate analysis results. The main factors directly or indirectly linked to the patient’s condition that are able to All research procedures were approved by the Research and change the results of AES analysis are the following: medications, Ethics Committee of UFJF - Hospital Universitário - Centro de vitamins, physical exercises, diet and poor sample storage(2, 5). Atenção à Saude (number: 0085.0.420.000-10). The laboratory has external quality control according to the standards of the Many situations do not allow the analysis within two hours Brazilian Society of Clinical Pathology (Sociedade Brasileira de after micturition, therefore the preservation of urine sample Patologia Clínica). becomes a commonly applied pre-analytical step in order to avoid the interferences previously mentioned. There are several preservatives that offer advantages and disadvantages: boric acid, Storage chloroform, phenol, formalin, toluene, thymol and refrigeration After performance of ACS test at immediate arrival, the (2, 7, 13) at 2 to 8°C, which is the most widely applied method due remaining urine volume (70 ml) was split into two vials that were to its cost-effectiveness and fewer drawbacks in comparison with stored for 24 hours following different procedures: one was stored chemical preservatives(2, 12, 13). in the refrigerator and the other was kept at room temperature. The use of refrigeration may lead to changes in ACS that The refrigerator temperature was adjusted between 2 to 8°C with include density, precipitation of crystals and even cellular time check by maximum and minimum thermometer throughout alterations(2, 13, 15). Moreover, the length period of cooling efficiency the experiment. ACS test was performed again. is controversial, insofar as some authors claim to be 12 hours(2), while others recommend up to 24 hours(12). ACS Once the material was collected, the aliquot of 10 ml of urine OBJECTIVES was applied for analysis. Physical examination was carried out by observing color, density, appearance and odor. The density was The present study aimed at verifying the influence of assessed with the use of a refractometer, which was subjected to refrigeration at 2 to 8ºC on urine storage for 24 hours before ACS calibration with distilled water after each analysis. 416 Karen Cristina Barcellos Ribeiro-; Bruno Rotondo Levenhagem Serabion; Eduardo Lima Nolasco; Chislene Pereira Vanelli; Harleson Lopes de Mesquita; José Otávio do Amaral Corrêa The chemical analysis was performed with the aid of and the remaining 1 ml was homogenized and placed in Newbauer reagent strips (UriquestPlus®) in order to detect the presence of chamber. Afterwards, pyocytes and red blood cells were counted leukocytes, nitrite, protein, blood, bilirubin, urobilinogen, ketone as described in the literature(17). The result was released with the body, glucose as well as pH control. After homogenization, the number of red blood cells or pyocytes/ml. Both ACS test and the reagent areas were immersed in fresh urine and immediately quantification of pyocytes
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