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n Medical Technology SA - A comparison of animal haemoglobins using high-performance liquid chromatography (HPL C) : scientific report
Automated cation-exchange HPLC is now considered the cornerstone diagnostic method in many laboratories for two reasons.
Firstly, unlike other methodologies it detects variant haemoglobins and measures haemoglobin's A, A2 and F percentages in a single procedure.
Secondly it is suitable for larger workloads in that it is now fully automated.
Haemoglobin is most generally characterised as consisting of two α and two non-α dimers that couple to form a tetramer in certain physiological conditions. This with its own haem group in a 'pocket' is then formed to make up a haemoglobin molecule.
This study was undertaken in order to document these different animal haemoglobins using HPLC in samples obtained from animal species in both the Kruger National Park and the Pretoria Zoo, with the proviso of setting up a comprehensive atlas of these particular animal haemoglobins, which could be used in a clinical diagnostic setting.
HPLC analysis was carried out on the following adult species:
Gorilla, chimpanzee, chacma baboon, vervet monkey, lion, leopard, white rhino, giraffe, wild dog, buffalo, blue and black wildebeest, Nile crocodile, eland, pigmy hippopotamus and cheetah. The findings indicated that each species had its own unique chromatogram when compared to that of humans, with regards to:Retention times (i.e. the time from application to elution from the column), their characteristic percentage and in some cases, in the shape of the peak or curve produced.
Analysis of these animal haemoglobins produced chromatograms with three or more significant peaks. Mostly these peaks corresponded to the α-globin chain, the β-globin chain and the haem unit. As confirmed in previous studies there were some species that had as many as four distinct chromatographic peaks.
The retention times produced from the 17 animal species examined in this study can then serve as profiles, which could be used to compile a comprehensive atlas of their unique haemoglobin chromatograms. It could also be used to identify each species where each has its own unique chromatogram. This atlas, similar to that compiled for the human haemoglobins could then be used in a clinical setting where abnormalities in the HPLC retention times may be detected if the animal is in any way compromised.
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