Effect of storage on biochemical parameters of packed red blood cells of goats in............ 257-261 citrate phosphate dextrose adenine/ saline adenine glucose mannitol

A study was conducted to assess the suitability of citrate phosphate dextrose adenine / saline adenine glucose mannitol as a storage media for packed RBCs of goats. Samples collected from ten apparently healthy goats were utilized for the study. Biochemical studies were carried out on day 0, 14, 28 and 42 days of storage using the parameters, viz. pH, glucose, potassium, malondialdehyde and reduced glutathione. The pH was stable throughout the study, whereas glucose showed significant reduction. Rest of the parameters increased significantly from 0th day to 42nd day. Based on the results, the storage media can be considered to be suitable for storing caprine packed RBCs.

Time Dependent Erythrocyte Morphometric Changes of Prolonged Stored Blood and its Effect on Target Post-transfusion Haematocrit of Splenectomised Mongrel Dogs

Background: Mean values of erythrocytic morphometric parameters of very old blood and its effect on the target post-transfusion haematocrit changes of splenectomised dogs was studied. Methods: Two hundred and fifty milliliters of blood each were drawn from healthy dogs (n=6) into citrate phosphate dextrose adenine-1 anti-coagulated blood bags, preserved for 35 days for the evaluation of erythrocyte morphometric and viability parameters. Thereafter, twenty adult male splenectomised dogs were randomly assigned into 5 groups (n=3). Post-splenectomy, 4, 14, 21 and 28 day old blood (DOB) were transfused to groups II-V while group I animals were not transfused. Intraoperative blood loss was determined during the surgery while post-transfusion, animals haematocrit were assayed and used to calculate the targeted haematocrit. Result: Findings revealed irreversible progressive time dependent morphometric changes by day 14 of blood storage. Hence, it is recommended that for transfusion purposes, 4 DOB should be the hallmark as it achieved the desired haematocrit and no morphometric changes were observed from it.

Determination of ceftriaxone in human plasma using liquid chromatography–tandem mass spectrometry

Ceftriaxone is a cephalosporin antibiotic drug used as first-line treatment for a number of bacterial diseases. Ceftriaxone belongs to the third generation of antibiotics and is available as an intramuscular or intravenous injection. Previously published pharmacokinetic studies have used high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV) for the quantification of ceftriaxone. This study aimed to develop and validate a bioanalytical method for the quantification of ceftriaxone in human plasma using liquid chromatography followed by tandem mass spectrometry (LC-MS/MS). Sample preparation was performed by protein precipitation of 100 µl plasma sample in combination with phospholipid-removal techniques to minimize matrix interferences. The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C18 column with 10 mM ammonium formate containing 2% formic acid: acetonitrile as mobile phase at a flow rate of 0.4 ml/min with a total run time of 10 minutes. Both the analyte and cefotaxime (internal standard) were quantified using the positive electrospray ionization (ESI) mode and selected reaction monitoring (SRM) for the precursor-product ion transitions m/z 555.0→396.1 for ceftriaxone and 456.0→324.0 for cefotaxime. The method was validated over the concentration range of 1.01-200 μg/ml. Calibration response showed good linearity (correlation coefficient > 0.99) and matrix effects were within the ±15% limit in 6 different lots of sodium heparin plasma tested. However, citrate phosphate dextrose plasma resulted in a clear matrix enhancement of 24% at the low concentration level, which was not compensated for by the internal standard. Different anticoagulants (EDTA, heparin and citrate phosphate dextrose) also showed differences in recovery. Thus, it is important to use the same anticoagulant in calibration curves and clinical samples for analysis. The intra-assay and inter-assay precision were less than 5% and 10%, respectively, and therefore well within standard regulatory acceptance criterion of ±15%.

Determination of carboxyhaemoglobin and methaemoglobin levels in donated blood

To determine the percentages of carboxyhaemoglobin (COHb) and methaemoglobin (MeHb) in donor blood and to compare these levels between smokers and nonsmokers at different time points during blood storage. Blood donors were recruited from Haematology Service, University Hospital Alzira Velano, Alfenas-MG. The blood was kept in collecting ducts (noodles) containing citrate, phosphate and dextrose (CPD) and stored at 4°C throughout the storage period. Since the noodles kept the characteristics of the bags, COHb and MeHb levels were analysed on the day of donation and after 20 days of storage. Levels of COHb and MeHb were determined using spectrophotometric methods. Non-parametric Friedman and Mann-Whitney tests were employed to compare COHb and MeHb levels before and after the storage and groups of smokers and nonsmokers, respectively. Levels of COHb and MeHb in the blood collected from smokers and nonsmokers were statistically different (p< 0.05; Mann- Whitney test) when the samples were analyzed before the storage. In blood of smokers, COHb levels were no different over a 20-day storage period (p= 0.7009; Friedman test). On the other hand, MeHb levels were significant different over a 20-day storage period (p< 0.05). The results suggest the need to regularly assess COHb and MeHb levels in donor blood stored in blood banks.

Cytoskeletal changes in Erythrocytes during storage in banked blood

Introduction: Erythrocytes have flexible, non-nucleated bi-concave shape with lipid bilayer cytoskeleton. Any alterations of erythrocyte shape make it susceptible to hemolysis. Blood for transfusion purpose is routinely stored in Citrate Phosphate Dextrose Adenine (CPDA-1) containing blood bags. During storage, blood undergoes an array of different morphological changes termed as “storage lesions” which makes it more fragile. This study was aimed to determine the structural and functional modifications in erythrocytes in CPDA-1 blood stored in local blood bank of KPK. Material & Methods: Blood from twenty healthy volunteer donors was taken and kept in CPDA-1 containing blood bags at Institute of Basic Medical Sciences (IBMS), Khyber Medical University (KMU). Hb-levels and Erythrocyte, Reticulocyte counts, Mechanical Fragility Index (MFI) and immunofluorescence staining for ankiyrin1 protein were performed on fresh blood samples. Samples for reticulocyte count was taken for 5 consecutive days while for the remaining parameters, blood was taken at 5 days interval till day 20th. The light and fluorescence micrographs were obtained accordingly and osmotic fragility tests were performed. Results: A significant mean reduction in erythrocyte counts and Hb-level was observed from day 0 to 20 (p=0.001), while MFI increased from day 0 to day 20 (12.88%±7.58 p=0.001). Reticulocyte count also decreased from day 0 up to day 5 (p=0.001). A weak association was observed between changes in MFI and erythrocyte morphology from day 0 to 20 (r=0.349), while the intensity and pattern of ankyrin1 protein expression appeared to change from day 10. Conclusion: Blood stored for a week has same properties as fresh blood, however, important structural alterations start to appear after the first week of storage and worsen with time. Therefore, to gain better transfusion results, blood stored for up to one week can safely be transfused.