Optimisation of pyrogen testing in parenterals according to different pharmacopoeias by probabilistic modelling

Abstract

The rabbit test to detect pyrogenic contamination in parenterals is crucial to ensure patient safety. The pharmacopoeial tests in Europe, the US and Japan are based on the fever reaction of rabbits, but differ in their experimental design and in their algorithms to assess contamination. Employing an international reference endotoxin, fever can be induced in rabbits. Data from 171 rabbits built the base for probabilistic modelling of the fever reaction and for the comparison of the pharmacopoeial tests. The rabbit fever reaction could be modelled as a function of the amount of injected endotoxin (per kg body weight) by linear regression. Combining the pharmacopoeial algorithms of the rabbit pyrogen test with the developed model allowed analysis of differences regarding test results and animal consumption. This showed that the assessment of pyrogenic contamination strongly depends on the respective pyrogen test stipulated by regulations. Additionally, the approach was used to develop a new experimental design. Two specific versions of this design resulted in a reduction of the number of animals used by about 30% while the safety of the test was maintained. A need for harmonisation is evident, allowing optimisation of the experimental design, which promotes animal welfare.

Keywords

Animal welfare animal model statistical model pharmacopoeias pyrogens

References

Poole S. , Dawson P. , Gaines Das RE Second international standard for endotoxin: calibration in an international collaborative study. J Endotoxin Res 1997; 4: 221—231.

Poole S. , Mussett MV The International Standard for Endotoxin: evaluation in an international collaborative study. J Biol Stand 1989; 17: 161—171.

Weary M. , Pearson III FC , Bohon J. , Donohue G. The activity of various endotoxins in the USP rabbit test and in three different LAL tests. Prog Clin Biol Res 1982; 93: 365—379.

Mascoli CC , Weary ME Limulus amebocyte lysate (LAL) test for detecting pyrogens in parenteral injectable products and medical devices: advantages to manufacturers and regulatory officials . J Parenter Drug Assoc 1979; 33: 81—95.

Mascoli CC , Weary ME Applications and advantages of the Limulus amebocyte lysate (LAL) pyrogen test for parenteral injectable products. Prog Clin Biol Res 1979; 29: 387—402.

The United States Pharmacopeial Convention I. Biological Test — Pyrogen Test. The United States Pharmacopoeia 24, Rockville, MD: 2000.

Council of Europe. Biological Tests, 2.6.8 Pyrogens. In: European Pharmacopoeia. Strasbourg : Council of Europe, 2001.

Society of Japanese Pharmacopoeia. General Tests, Pyrogen Test. The Japanese Pharmacopoeia (English version). Tokyo: 1996.

Russel Wms , Burch RL The Principles of Humane Experimental Technique. London : Methuen, 1959.

10.

Hutchinson TH , Barrett S. , Buzby M. et al. A strategy to reduce the numbers of fish used in acute ecotoxicity testing of pharmaceuticals. Environ Toxicol Chem 2003; 22: 3031—3036.

11.

Stallard N. , Whitehead A. , Indans I. Statistical evaluation of an acute dermal toxicity test using the dermal fixed dose procedure. Hum Exp Toxicol 2004; 23: 405—412.

12.

Stallard N. , Whitehead A. , Indans I. Statistical evaluation of the fixed concentration procedure for acute inhalation toxicity assessment. Hum Exp Toxicol 2003; 22: 575—585.

13.

Diener W. , Kayser D. , Schlede E. The dermal acute toxic class method: test procedures and biometric evaluations . Arch Toxicol 1998; 72: 751—762.

14.

van den Heuvel MJ , Clark DG , Fielder RJ et al. The international validation of a fixed-dose procedure as an alternative to the classical LD50 test. Food Chem Toxicol 1990; 28: 469—482.

15.

Schlede E. , Mischke U. , Diener W. , Kayser D. The international validation study of the acute toxic class method (oral) . Arch Toxicol 1995; 69: 659—670.

16.

Bruce RD A confirmatory study of the up-and-down method for acute oral toxicity testing . Fundam Appl Toxicol 1987; 8: 97—100.

17.

Weisser K. , Hechler U. Animal Welfare Aspects in the Quality Control of Immunobiologicals. Nottingham: FRAME, 1997.

18.

Hartung T. , Aaberge I. , Berthold S. et al. Novel pyrogen tests based on the human fever reaction. The report and recommendations of ECVAM Workshop 43. European Centre for the Validation of Alternative Methods. European Centre for the Validation of Alternative Methods. Altern Lab Anim 2001; 29: 99—123.

19.

Dabbah R. , Ferry Jr E. , Gunther DA et al. Pyrogenicity of E. coli O55:b5 endotoxin by the USP rabbit test — a HIMA collaborative study. J Parenter Drug Assoc 1980; 34: 212—216.

20.

Council of Europe. General Texts, 5.8. Pharmacopoeial Harmonisation. In: European Pharmacopoeia . Strasbourg: Council of Europe , 2001.

21.

Twohy CW , Duran AP , Munson TE

Endotoxin contamination of parenteral drugs and radiopharmaceuticals as determined by the Limulus amebocyte lysate method. J Parenter

Sci Technol 1984; 38: 190—201.

22.

Oecd. OECD Guideline for Testing of Chemicals No. 420: Acute Oral Toxicity — Fixed Dose Procedure. 1—14. Paris: Organisation for Economic Cooperation and Development, 2001.

23.

Oecd. OECD Guideline for Testing of Chemicals No. 423: Acute Oral toxicity — Acute Toxic Class Method. 1-14. Paris: Organisation for Economic Cooperation and Development, 2001.

24.

Oecd. OECD Guideline for Testing of Chemicals No. 425: Acute Oral Toxicity: Up-and-Down Procedure. 1-26. Paris: Organisation for Economic Cooperation and Development, 2001.

25.

Oecd. Guidance Document on Acute Oral Toxicity Testing. 1-24. Environmental Health and Safety Monograph Series on Testing and Assessment No. 24. Paris: Organisation for Economic Cooperation and Development , 2001.

26.

Tschumi J.

Comparison of temperature rise interpretations between European and United States Pharmacopeias' pyrogen tests. PDA

J Pharm Sci Technol 2003; 57: 218—227.