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

Bhomia, Ruchir, Trivedi, Vivek, Coleman, Nichola, Mitchell, John C. (2016) The thermal and storage stability of bovine haemoglobin by ultraviolet–visible and circular dichroism spectroscopies. Journal of Pharmaceutical Analysis, (6). pp. 242-248. ISSN 2095-1779. E-ISSN 2214-0883. (doi:10.1016/j.jpha.2016.02.004)

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 than at 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 decreased steadily with the increasing temperature above 35 °C at all pH values. The loss in α-helicity and gain in random coil conformations was pH-dependent and the 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 elucidates the significance of storage and processing conditions on its stability.

Item Type: Article
DOI/Identification number: 10.1016/j.jpha.2016.02.004
Uncontrolled keywords: Bovine haemoglobin, Circular dichroism, Thermal stability, Storage stability, Ultraviolet--visible spectroscopy
Subjects: R Medicine > RS Pharmacy and materia medica
Divisions: Central Services > Universities at Medway
Faculties > Sciences > Medway School of Pharmacy
Depositing User: Vivek Trivedi
Date Deposited: 24 Apr 2019 10:28 UTC
Last Modified: 03 Jun 2019 09:43 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/73605 (The current URI for this page, for reference purposes)
Trivedi, Vivek: https://orcid.org/0000-0001-9304-9214
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