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The thermal and storage stability of bovine haemoglobin by ultra violet–visible and circular dichroism spectroscopies

The thermal and storage stability of bovine haemoglobin by ultra violet–visible and circular dichroism spectroscopies

Bhomia, Ruchir, Trivedi, Vivek ORCID logoORCID: https://orcid.org/0000-0001-9304-9214, Coleman, Nichola J. and Mitchell, John C. ORCID logoORCID: https://orcid.org/0000-0003-2945-3292 (2016) The thermal and storage stability of bovine haemoglobin by ultra violet–visible and circular dichroism spectroscopies. Journal of Pharmaceutical Analysis, 6 (4). pp. 242-248. ISSN 2095-1779 (Print), 2214-0883 (Online) (doi:10.1016/j.jpha.2016.02.004)

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Abstract

The effects of temperature, pH and long-term storage on the secondary structure and conformation changes of bovine haemoglobin (bHb) were studied using circular dichroism (CD) and ultraviolet-visible (UV-vis) spectroscopies. Neural network software was used to deconvolute the CD data to obtain the fractional content of the five secondary structures. The storage stability of bHb solutions in pH 6, 7 and 8 buffers was significantly higher at 4 °C in comparison to 23 °C for the first 3 days. A complete denaturation of bHb was observed after 40 days irrespective of storage temperature or pH. The bHb solutions were also exposed to heating and cooling cycles between 25 and 65 °C and structural changes were followed by UV-vis and CD spectroscopies. These experiments demonstrated that α-helix content of bHb decreases steadily with increasing temperature above 35 °C at all pH values. The loss in α-helicity and gain in random coil conformations was pH-dependent and greatest under alkaline conditions. Furthermore, there was minimal recovery of the secondary structure content upon cooling to 25 °C. The use of bHb as a model drug is very common and this study illustrates the significance of storage and processing conditions on its stability.

Item Type: Article
Additional Information: © 2016 Xi'an Jiaotong University. Production and hosting by Elsevier B.V. This is an open access article under the CCBY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Uncontrolled Keywords: Bovine haemoglobin, Circular dichroism, Thermal stability, Storage stability
Faculty / School / Research Centre / Research Group: Faculty of Engineering & Science
Faculty of Education, Health & Human Sciences > School of Human Sciences (HUM)
Last Modified: 09 Oct 2021 04:45
URI: http://gala.gre.ac.uk/id/eprint/15256

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