Investigation of the Effects Stored Blood for Transfusion has on the Haemolysis of Red Blood Cells
Cardiff Metropolitan University
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Aim The aim of this project is to determine if the duration red blood cells are stored for effects the amount of haemolysis present at the end of storage. Methods Blood was taken from 6 different units of blood. They were centrifuged at 3000rpm for 10 minutes. The supernatant was removed and transferred to a fresh tube and centrifuged for 5 minutes at 3000rpm. The supernatant was then transferred into tubes and stored in the freezer. A stock solution for the haemoglobin reagent was made by dissolving sodium dodecyl sulphate (SDS) in phosphoric acid buffer and Triton X-100 was added to this16. The phosphoric acid buffer was added to make the stock solution up to 1000ml16. A working solution was made by diluting the stock solution 1:100 using deionized water16. Calibration standards were made by doubling dilutions starting with 140g/dL. Absorbance’s were recorded at 539nm. 20μl of blood was mixed with 5ml of the working solution by gently shaking16. Absorbance’s were read at 539nm16. Summary of Results The R2 value for all the samples compared together was 26.3% this shows that all the units compared together do not fit closely to the regression line and the concentration of haemoglobin does not fit with the duration of storage time. The R2 value for units 2 and 5 (71.2% and 81.7%) show that there is a relationship between an increase in storage time and an increase in haemoglobin concentration. The R2 value for units 1, 3 and 4 (22.4%, 7.3% and 27.8%) show there is no relationship between an increase in storage time and an increase in haemoglobin concentration. The R2 value for unit 6 (57.5%) shows a slight trend in the relationship between an increase in storage time and an increase in haemoglobin but this could be because the concentration of haemoglobin stays unchanged for days 4, 8 and 11. The ANOVA test showed a P value of 0.011 meaning the means of the samples showed significant difference. Main Findings and Conclusions The findings show that some units represent the principle that an increase in storage effects the amount of haemolysis present. Units 2, 3 and 5 signify this hypothesis with an increase in haemolysis on the final day of storage compared to day 1. Units 1, 4 and 6 do not prove this hypothesis because units 4 and 6 show a decrease in overall haemolysis from day 1 to the last day of storage and unit 1 presents the same amount of haemolysis on day 1 and day 28.
BSc (Hons) Biomedical Science
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