Variability of blood biochemical parameters and establishing of reference ranges in nonclinical studies. Part 1: rats

DOI: 10.29296/2618723X-2020-01-06

N.G. Voitenko, M.N. Makarova, A.A. Zueva

Scientific-Production Organisation «Home of Pharmacy»

188663, Russia, Leningrad oblast, Vsevolozhskiy district, Kuzmolovskiy t.s., Zavodskaya st. 3-245

Е-mail:  [email protected]

Keywords: reference intervals of blood biochemical parameters retrospective analysis rats

For citation:

Voitenko N.G, Makarova M.N., Zueva A.A. Variability of blood biochemical parameters and establishing of reference ranges in nonclinical studies. Part 1: rats. Laboratory Animals for Science. 2020; 1.


Establishing of current reference ranges for laboratory animal parameters is topical problem both in clinical and nonclinical studies. Retrospective analysis of 11 serum biochemical parameters in rats was conducted. Blood samples were collected from 195 male and 184 female rats (age 12–20 weeks and body weight 250–350 g.).

It was found that outliers of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were identified most frequent than other blood biochemical parameters in rats. This fact requiring increase in the number of repeatability in analyzing of blood parameters has to be taken into account when planning of study.

Reference ranges of data base including creatinine, urea, AST, ALT, alkaline phosphatase, cholesterol, triglycerides, total protein, albumin, glucose and total bilirubin and its’ inter-individual variance were identified. It was determined that alkaline phosphatase, triglycerides, glucose and total bilirubin have greatest inert-individual variance (more than 30%).

The data have been received from retrospective analysis was compared with reference interval of rat biochemical parameters from three large sources (Charles River, Taconic and Envigo). Reference intervals from literature sources also indicate high variance of some enzyme activity (including alkaline phosphatase), glucose, total bilirubin and triglycerides. Our calculated reference intervals was comparable with the same data from literature sources.

The results indicate that retrospective data analyzing allowing to access more accurate reference interval from larger data base without compromising ethical principles is preferable to use. Comparative analysis of inter-individual variance of rat and human blood biochemical parameters show the presence of species differences which need should be considered in discussion the preclinical study results.

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  1. Теория статистики: учебное пособие. Л.Г. Батракова. М.: КНОРУС, 2016; 528 [Teoriya statistiki: uchebnoe posobie / L.G. Batrakova. M.: KNORUS, 2016. 528 p. (in Russ.)].
  2. ГОСТ Р53022-3–2008. Технологии лабораторные и клинические. Требования к качеству лабораторных исследований. Правила оценки клинической эффективности лабораторных тестов. М., 2008. [GOST R53022-3–2008. Tekhnologii laboratornye i klinicheskie. Trebovaniya k kachestvu laboratornyh issledovanij. Pravila ocenki klinicheskoj effektivnosti laboratornyh testov. M., 2008 (in Russ.)].
  3. ГОСТ Р 53022.2–2008. Технологии лабораторные клинические. Требования к качеству клинических лабораторных исследований. Часть 2. Оценка аналитической надежности методов исследования (точность, чувствительность, специфичность) М., 2008. [GOST R 53022.2–2008 Tekhnologii laboratornye klinicheskie. Trebovaniya k kachestvu klinicheskih laboratornyh issledovanij. CHast' 2. Ocenka analiticheskoj nadezhnosti metodov issledovaniya (tochnost', chuvstvitel'nost', specifichnost') M., 2008. (in Russ.)].
  4. Horowitz G.L., Altaie S., Boyd J.C., Ceriotti F., Garg u., Horn P., Pesce A., Sine H.E., Zakowski J. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline. 3-rd Ed. Clinical and Laboratory Standards Institute. 2008, 28(30): 1–59.
  5. Mazzachi B.C., Peake M.J., Ehrhardt V. Reference range and method comparison studies for enzymatic and Jaffé creatinine assays in plasma and serum and early morning urine. Clinical laboratory. 2000, 46 (1–2): 53–5.
  6. Junge W., Wilke B., Halabi A., Klein G. Determination of reference intervals for serum creatinine, creatinine excretionand creatinine clearance with an enzymatic and a modified Jaffé method. Clinica Chimica Acta. 2004, 344 (1–2): 137–48. DOI: 10.1016/j.cccn.2004.02.007
  7. Burtis C.A., Ashwood E.R., Bruns D.E. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics 4th Edition. MO: Elsevier Saunders, 2005: 2448 p. DOI: 10.1373/clinchem.2005.062638
  8. Конвертация традиционных единиц измерения используемых в лабораторной и медицинской практике в единицы международной системы СИ. URL:
  9. CDF (Fisher) rat hematology and biochemistry. URL:
  10. Lewis rat hematology and biochemistry. URL:
  11. Clinical laboratory parameters for Crl:CD(SD) Rats. URL:
  12. Clinical laboratory parameters for Crl:WI(Han) Rats. URL:
  13. Fisher 344 Inbred. Phenotypic data. URL:
  14. Lewis Inbred. Phenotypic data. URL:
  15. Sprague Dawley outbred. Phenotypic data. URL:
  16. Wist@Tac (GALAS) Outbred
  17. Wistar. Hsd:WI. 7-8 weeks old, Males. URL:
  18. Wistar. Hsd:WI. 7-8 weeks old, Females. URL:
  19. Sprague Dawley. Hsd:Sprague Dawley®SD®. 6-8 weeks old, Males. URL:
  20. Sprague Dawley. Hsd:Sprague Dawley®SD®. 6-8 weeks old, Females. URL:
  21. Wistar Han. RccHan®:WIST. 6-8 weeks old, Males. URL:
  22. Wistar Han. RccHan®:WIST. 6-8 weeks old, Females. URL:
  23. Lewis. LEW/SsNHsd. 11-12 weeks old, Males. URL:
  24. Caisey J.D., King D.J. Clinical chemical values for some common laboratory animals. Clinical Chemistry. 1980, 26 (13): 1877–9.
  25. Близнецова Г.Н., Брюхова И.В. Влияние препарата «Прималакт» на гематологические и биохимические показатели крови лабораторных животных. Вестник Воронежского государственного аграрного университета. 2013, Т. 4: 233–5. [Bliznecova G.N., Bryuhova I.V. Vliyanie preparata primalakt na gematologicheskie i biohimicheskie pokazateli krovi laboratornyh zhivotnyh. Vestnik Voronezhskogo gosudarstvennogo agrarnogo universiteta. 2013, T. 4: 233–5 (in Russ.)].

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