Validation of the biochemical method of determining the ammonia level in the biological liquid samples as a tool for providing the quality of hyperamonemia diagnostics
Keywords:validation, determination procedure, ammonia, convergence and reproducibility, correctness and uncertainty of measurements
Nowadays, one of the most topical issues in the healthcare system ofUkraineis to ensure the quality of laboratory research. For this purpose, it is necessary to develop and implement a quality system in accordance with ISO 15189: 2015 “Medical laboratories. Basic requirements for quality and competence”, its significant stage is conducting validation. The main objects are the methods by which measurements are made in the laboratory. In case of liver lesions, a laboratory examination is mandatory, including the study of the pigment (total, direct and indirect bilirubin), enzyme (ALT, AST, LDH, alkaline phosphatase, etc.) and protein metabolism. The liver also acts as a central organ in the metabolism of nitrogen compounds; in it a significant amount of ammonia is formed and neutralized. Therefore, the level of blood ammonia is an important parameter that reflects the functional state of the liver and can serve as an indicator of the severity of damage to this organ.
Aim. To develop the method for conducting validation procedures in order to assess the suitability of the biochemical method for determining the level of ammonia in biological liquids in the Laboratory of Clinical Diagnostics of the Clinical Diagnostic Center of the NUPh.
Materials and methods. The object of the study was a standardized method for determining the level of ammonia. An “Ammonia NH3” reagent manufactured by Randox, USA, and the known concentration of “Ammonia ethanol control – level2” manufactured by Randox, USA as a control serum were selected for the study. The measurements were carried out on an Express Plus automatic biochemical analyzer manufactured by Bayer Corporation,Germany. When processing the research results the descriptive statistics indicators were used, and a number of statistical evaluations were carried out.
Results. The validation procedure consists of several stages. At the premises of the Laboratory of Clinical Diagnostics of the CDC of the NUPh the protocol and the validation report were developed to assess the suitability of the method for determining the concentration of ammonia in biological liquids by the enzymatic UV method on an Express Plus automatic biochemical analyzer. Validation was carried out using reagents and control material manufactured byRandox,USA. The following validation characteristics of the method were determined: specificity, convergence and reproducibility, correctness, measurement uncertainty. The assessment of the intermediate convergence and reproducibility of the ammonia determination method when working on the analyzer by two laboratory assistants of the same qualification indicates the absence of gross errors in the operation of the device and statistically important differences during measurements. The assessment of the method correctness using the control material has proven that the systematic error is not significant (according to the given criterion of acceptability). Thus, this method provides the accuracy of measurements within the laboratory. The calculation of the expanded uncertainty has shown that the obtained values of the ammonia level can be considered accurate and reliable.Conclusions. The validation of the method for determining ammonia in the human blood by the enzymatic method has proven that this method has performance characteristics that meet the regulated ones, satisfies the established criteria, and the parameters measured correspond to the appropriate ones.
Tapper, E. B. (2019). Predicting Overt Hepatic Encephalopathy for the Population With Cirrhosis. Hepatology, 70(1). https://doi.org/10.1002/hep.30533
Redant, S., Beretta-Piccoli, X., Mugisha, A., Attou, R., Kaefer, K., De Bels, D., … Honoré, P. M. (2019). Hyperammonemia, the Last Indication of High-Volume Hemodiafiltration in Adult and Children: A Structured Review. Blood Purification, 48(4), 330–335. https://doi.org/10.1159/000501390
Mpabanzi, L., Olde Damink, S. W. M., van de Poll, M. C. G., Soeters, P. B., Jalan, R., & Dejong, C. H. C. (2011). To pee or not to pee. European Journal of Gastroenterology & Hepatology, 23(6), 449–454. https://doi.org/10.1097/meg.0b013e328346a7bd
Laleman, W., Verbeke, L., Meersseman, P., Wauters, J., van Pelt, J., Cassiman, D., … Nevens, F. (2011). Acute-on-chronic liver failure: current concepts on definition, pathogenesis, clinical manifestations and potential therapeutic interventions. Expert Review of Gastroenterology & Hepatology, 5(4), 523–537. https://doi.org/10.1586/egh.11.47
Bueverov, A. O. (2015). Medicinskij sovet, 13, 80–85.
Zupanets, I. A., Popov, S. B., Rudik, Yu. S., KotvItskа, A. A. (2018). Simptomi ta sindromi v praktichnIy farmatsii. Kharkiv: NPhaU: Zoloti storinky, 49–50.
NAKAZ vid 1 zhovtnia 2015 roku N 644. (2015). Pro zatverdzhennia Kontseptsii systemy menedzhmentu yakosti v medychnykh laboratoriiakh Ukrainy u vidpovidnosti do vymoh mizhnarodnykh standartiv ta Planu zakhodiv shchodo yii realizatsii. Available at: https://ips.ligazakon.net/document/MOZ25118
Analizy pri zabolevaniiakh pecheni. (n.d.). Available at: https://www.kp.ru/guide/analizy-pri-zabolevanijakh-pecheni.html
9. Kontrol kachestva laboratornykh yssledovanyi. (2015). Zdorov’ia Ukrainy Infomedia. Available at: https://health-ua.com/article/16468-kontrol-kachestva-laboratornyh-issledovanij
Dobrova, V. Ye. (2010). Teoretychni aspekty planuvannia vymiriuvalnykh eksperymentiv pry provedenni klinichnykh doslidzhen likarskykh zasobiv. Management, Economics and Quality Assurance in Pharmacy, 3(11), 6–11.
Ratushna, K. L., Mazur, N. S., Zupanets, K. O., Dobrova, V. Ye. (2013). Zabezpechennia yakosti biokhimichnykh doslidzhen v systemi upravlinnia danymy pry klinichnykh vyprobuvanniakh. Ukrainian biopharmaceutical journal, 4(27), 17.
Misiurova, S. V., Dobrova, V. Y., Svid, N. O., & Otrishko, I. A. (2018). Implementation of the quality system according to ISO 15189:2015 in the Laboratory of Clinical Diagnostics of the ClinicalDiagnosticCenterat the National University of Pharmacy: applied aspects. Clinical pharmacy, 22(1), 28–33. https://doi.org/10.24959/cphj.18.1449
Derzhavna farmakopeia Ukrainy. (1-st edition). (2001). Derzhavne pidpryiemstvo «Naukovo-ekspertnyi farmakopeinyi tsentr». Kharkiv: RERIH, 556.
14. ISO 15189–2009–40. (n.d.). Laboratorіi medychnі. Okremі vymohi do yakostі ta kompetentnostі.
Mіsiurova, S., Dobrova, V., Svid, N., Otrishko, I., & Sakharova, T. (2018). Quality assurance of biochemical studies using validation procedures in the laboratory of clinical diagnostics of the clinical diagnostic center of the national university of pharmacy on the example of the method of determining the level of general and diirect bilirubin in biological fluids. Clinical Pharmacy, 22(3), 52–57. https://doi.org/10.24959/cphj.18.1468
Dobrova, V. Ye., Kovalenko, S. M., Maloshtan, L. M, Stepanova, K. O. (2009). Validatsiia bioanalitychnykh metodiv dlia farmakolohichnykh doslidzhen likarskykh zasobiv zghidno vymoh standartu DSTU ISO/IES 17025:2005. Management, Economics and Quality Assurance in Pharmacy, 4(6), 9–13.
Misiurova, S., Dobrova, V., Svid, N., Otrishko, I., & Propisnova, V. (2019). Approaches to the implementation of the quality system in accordance with ISO 15189:2015 in the Laboratory of Clinical Diagnostics of the Clinical and DiagnosticCenterat the National University of Pharmacy on the example of methods for determining the levels of lactate dehydrogenase and glucose in biological fluids. Clinical Pharmacy, 23(3), 12–18. https://doi.org/10.24959/cphj.19.1486
Dobrova, V. Ye, Zupanets, І. A. (2011). Teoretychni pidkhody do otsinky ta rozrakhunku nevyznachenostei rezultativ klinichnykh vyprobuvan. Clinical pharmacy, 15(4), 18–20.
Dobrova, V. Ye. (2010). Teoreticheskie aspekty planirovaniia izmeritelnykh eksperimentov pri provedenii klinicheskikh ispytanii lekarstvennykh sredstv. Systemy obrobky informatsii, 4(85), 145–147.
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