Reversed-phase high performance liquid chromatography as an alternative to animal bioassay for human thyrotropin potency determination

Author(s)

Almeida, B. E. ^[1] ; Damiani, R. ^[1] ; Oliveira, J. E. ^[1] ; Dalmora, S. L. ^[2] ; Torjesen, P. A. ^[3] ; Bartolini, P. ^[1] ; Ribela, M. T. C. P. ^[1]

Affiliation

^[1] Cidade Univ, IPEN CNEN, Dept Biotechnol, BR-05508900 Sao Paulo - Brazil

^[2] Univ Fed Santa Maria, Dept Ind Pharm, BR-97119900 Santa Maria, RS - Brazil

^[3] Oslo Univ Hosp, Hormone Lab, N-0424 Oslo - Norway

• Document type: Journal article
• Source: ANALYTICAL METHODS; v. 6, n. 17, p. 6688-6694, SEP 7 2014.
• Web of Science Citations: 8

Abstract

Reversed-phase high performance liquid chromatography (RP-H PLC) was compared to the in vivo bioassay (BA) based on TSH-induced T-4 determination in mice. A linear relationship (BA(mu g) = 0.9790 RP-HPLC mu g - 0.052) with a highly significant correlation (r = 0.87; p < 0.001; n = 14 preparations) was found between these two methods. The mean difference between the bioactivity predicted from RP-H PLC data via this equation and the mean of the bioactivities obtained with the two methods for 11 other hTSH preparations was -2.01%, with a 95% confidence interval of -13.13% to +9.11%. This analysis included 5 totally or partially-degraded samples, indicating a useful correlation between the two determinations and, as expected, a higher sensitivity of the physical-chemical method. Interestingly, a commercial hTSH tot did not show any alteration 2 years post expiration date. These results demonstrate that RP-H PLC is a novel and viable alternative to the use of in vivo bioassays for hTSH potency determination, thus avoiding or reducing animal use. (AU)

• FAPESP's process: 11/07289-0 - Synthesis and characterization of different glycoforms of recombinant human thyrotropin (rhTSH) pharmacologically actives
• Grantee: Maria Teresa de Carvalho Pinto Ribela
• Support Opportunities: Regular Research Grants