Background: Storage of whole blood has deleterious effects on the biochemical parameters of RBCs. Aim: To study the changes in biochemical parameters in whole blood upon storage. Materials and Methods: 30 healthy donors were included in this study and effect of storing whole blood was studied on day 0, 3, 7, 14 and 21. Results: Highly significant changes were noted in the levels of blood pH, sodium, potassium, chloride, bicarbonate, aspartate transaminase and lactate dehydrogenase. Conclusion: There are significant changes in the biochemical parameters during storage. The most significant change is the marked increase in potassium levels. In order to reduce the deleterious effects of stored blood in the recipient, it is advisable to check the clinical details of the patient. Care should be taken to issue fresh blood (not more than 7 days old) particularly to patients with renal disorders.

Bajaj, Sachin

Jagga, Kapil

Jha, Vidya

Tirkey, Rekha

English

Advanced Research Publications: Medical Journals

Research Article     © ADR Journals 2017. All Rights Reserved.  Biochemical Changes in Whole Blood -                          Effect of Storage Vidya Jha1, Rekha Tirkey2, Kapil Jagga3, Sachin Bajaj4 Abstract Background: Storage of whole blood has deleterious effects on the biochemical parameters of RBCs.  Aim: To study the changes in biochemical parameters in whole blood upon storage. Materials and Methods: 30 healthy donors were included in this study and effect of storing whole blood was studied on day 0, 3, 7, 14 and 21. Results: Highly significant changes were noted in the levels of blood pH, sodium, potassium, chloride, bicarbonate, aspartate transaminase and lactate dehydrogenase. Conclusion: There are significant changes in the biochemical parameters during storage. The most significant change is the marked increase in potassium levels. In order to reduce the deleterious effects of stored blood in the recipient, it is advisable to check the clinical details of the patient. Care should be taken to issue fresh blood (not more than 7 days old) particularly to patients with renal disorders. Keywords: Whole blood, CPDA, Storage, Biochemical changes Introduction Whole blood is stored at 4°C for a period of 35 days in the blood bank. Due to storage, there is alteration in the physical and biochemical properties of RBCs. These are known as storage lesions. Our body maintains pH and temperature of blood in circulation as well as continuously provides nutrients and flushes out the waste products.1 However, with time, RBCs in stored blood undergo deterioration in both mechanical and biochemical features.  Caraway compared certain biochemical tests in RBCs with that of plasma and found that erythrocytes had 20-fold as much aspartate aminotransferase, 160-fold lactate dehydrogenase, 67-fold as much acid phenyl phosphatase and 23-fold as much potassium as had plasma.2 Hemolysis of RBCs can affect the biochemical features to a wider extent, i.e., it can increase the serum levels of certain constituents released from RBCs due to hemolysis, decrease the levels owing to dilution, and hemoglobin released also can interfere with colorimetric measurements. Burns and co-workers defined visible hemolysis as an extracellular hemoglobin (Hb) concentration of 0.3 g/L (4.65 mol/L) that leads to a detectable pink-to-red color of serum or plasma with a visible appearance in blood samples with as low as 0.5% hemolysate.3 This study highlights the biochemical changes in stored blood. 1Senior Resident, Department of Pathology, 2Unit Incharge, 3,4MD Pathology, Blood Bank and Transfusion Medicine, Vardhman Mahavir Medical College and Safdarjang Hospital, New Delhi, India.  Correspondence: Dr. Vidya Jha, Vardhman Mahavir Medical College and Safdarjang Hospital, New Delhi, India. E-mail Id: vidyajha29@gmail.com Orcid Id: http://orcid.org/0000-0001-7524-3566 How to cite this article: Jha V, Tirkey R, Jagga K et al. Biochemical Changes in Whole Blood – Effect of Storage. Rec Adv Path Lab Med 2017; 3(1): 4-6. ISSN: 2454-8642 Rec. Adv. Path. Lab. Med. 2017; 3(1)          Jha V et al. 5  ISSN: 2454-8642 Materials and Methods 30 healthy volunteer donors were included in the study. 350 mL of blood was drawn from each donor into bags containing 49 mL citrate phosphate dextrose adenine anticoagulant (CPDA-1). We followed the guidelines of regulations of drugs and cosmetics rules as given by the Government of India for screening blood donors.4 Upon collection, blood bags were properly stored in a quarantine shelf at 2–4°C in the blood bank. Citrate phosphate dextrose adenine solution is known to extend the life of stored RBCs for 5 weeks.5 For various tests in the study, 5 mL of blood was taken from the stored bags and analyzed at 0, 3, 7, 14 and 21 days. Values obtained at day 0 were kept as control values. The biochemical tests done included pH, sodium, potassium, chloride, bicarbonate, calcium, aspartate transaminase, alanine transaminase and lactate dehydrogenase. Statistical analysis was done using SPSS software version 21.0 and p value <0.05 was considered statistically significant. Results  The 30 donors included in the study had age range between 20 and 54 years (mean 26 years). There were 28 males and 2 female donors. The most common blood group was found to be B+ (n=14), followed by O+ (n=10), A+ (n=4), AB+ (n=1) and B− (n=1). All the blood samples were negative for HIV, hepatitis B and C, malaria and syphilis. Table 1 shows the results of different biochemical tests done. Discussion The most commonly used anticoagulant for whole blood is CPDA-1. Its constituents are citrate, which chelates ionized calcium to prevent coagulation, phosphate containing anticoagulants as they have lower acidity than other anticoagulants and have a higher concentration of 2,3 DPG, and red cell phosphate, dextrose which is a source of energy for the RBCs, and adenine which is part of ATP and viability of RBCs is regenerated by addition of adenine after blood transfusion.4-6  It is due to the prolonged contact of plasma with RBCs that results in exchange of constituents between them. It results in variations in different analyte levels by concentrating or diluting the particular analyte. The red blood cells in stored blood bags have decreased viability owing to utilization of glucose and a subsequent ATP (adenosine triphosphate) depletion.7 This in turn reduces the activity of ATPase pump. Under physiological conditions, three sodium ions are pumped out of the cell in exchange of two potassium ions which are pumped in. Loss of this pumping activity leads to hyperkalemia and hyponatremia as was seen in our study. Similar finding of hyperkalemia was noted by Adias and his coworkers but they did not find any significant change in sodium ion.8 It is known that low temperature also induces reduction in ATPase activity which in turn leads to extracellular pumping of K+ and entry of Na+ ions into RBCs.8.9 Hemolysis also leads to hyperkalemia. All these factors must have contributed to the observed high rise in potassium levels in this study. There was a significant reduction in pH at day 21 in this study. Similarly anaerobic glycolysis causes increase in lactic acid concentration which causes a lowering of pH. LDH levels are known to reflect the degree of hemolysis as hemolysis results in release of LDH in plasma.2 LDH levels were found to be significantly raised in this study. With each 0.1 unit change in pH there is 0.4 mmol/L change in the serum potassium level. Potassium levels rise during acidosis and decrease during alkalosis.10 This also causes hyperkalemia. In our study we found a significant increase in the levels of potassium ions from day 0 to day 21. Such high levels can have deleterious effect on recipients, particularly with kidney disorders.  Low levels of ionized calcium were observed in the study as citrate present in the anticoagulant chelates it. As far as chloride levels are concerned, they showed a transient increase for first seven days followed by a significant decrease. This is because of the fact that sodium, chloride and calcium have low molecular weights and while being stored they enter RBCs owing to their concentration gradients.11  Similar result was seen for bicarbonate ions. On day 21, they showed a significantly reduced level. Bicarbonate ions are used to buffer excess of lactic acid produced by RBCs during anerobic metabolism. Moderate hemolysis also decreases bicarbonate levels. Latham et al. and Bailey et al. also observed decline in concentration of plasma glucose and bicarbonate and increase in potassium, lactate, LDH, ammonia, and hemoglobin concentration with storage.12,13 RBCs are known to have 20 fold as high concentration of AST as compared to plasma. Therefore, even a mild hemolysis will produce significant changes in AST. We observed statistically significant differences in AST levels from day 0 to day 21. This could be due to ongoing hemolysis in stored blood. Similar finding was noted by Koseoglu and coworkers.14 However, ALT levels were not increased significantly at the end of the study. This could be due to the fact that during intravascular hemolysis, AST is released more than ALT.15  Jha V et al.             Rec. Adv. Path. Lab. Med. 2017; 3(1) ISSN: 2454-8642  6 Table 1.Changes in Biochemical Parameters in Stored Whole Blood on Day 0, 3, 7, 14 and 21 (*significant p value) Parameters Day 0 Day 3 Day 7 Day 14 Day 21 p value pH 7.15±0.25 6.95±0.05 6.84±0.06 6.82±0.10 6.78±0.03 <0.0001* Sodium (mEq/L) 154.6±2.01 151.2±2.52 148±5.99 141.0±5.12 138±4.93 <0.0001* Potassium (mEq/L) 3.50±0.22 6.35±0.53 11.10±0.94 15.65±2.13 18.46±2.98 <0.0001* Chloride (mEq/L) 87.00±2.35 90.06±2.40 96.8±5.65 94.2±2.15 86.23±2.88 <0.0001* Bicarbonate (mg/dL) 17.5±1.22 16.53±1.83 13.01±2.56 6.63±3.21 4.82±1.5 <0.0001* Calcium (mg/dL) 0.41±0.004 0.042±0.005 0.046±0.04 0.049±0.02 0.05±0.002 >0.05 Aspartate transaminase (mg/dL) 26.14±5.56 28.23±6.53 30.12±8.96 41.24±10.62 45.57±4.28 <0.0001* Alanine transaminase (mg/dL) 38.8±12.83 41.02±20.70 38.34±21.8 48±24.26 49.5±23.56 >0.05 Lactate dehydrogenase (mg/dL) 234±35.6 297±46 490±123.2 577.3±80.4 675±52 <0.001*  Conclusion  The most commonly used anticoagulant for storing whole blood is CPDA. However, there are significant changes in the biochemical parameters during storage. The most significant change is the marked increase in potassium levels. In order to reduce the deleterious effects of stored blood in the recipient, it is advisable to check the clinical details of the patient. Care should be taken to issue fresh blood (not more than 7 days old), particularly to patients with renal disorders. However, further studies might be done to consolidate these findings.  Conflict of Interest: None  References  1. Rudmann SV. Textbook of blood banking and transfusion medicine. Blood component preservation and storage. (2nd edn). Elsevier Health Sciences. 2005; 269. 2.  Frank JJ, Bermes EW, Bickel MJ et al. Effect of in vitro hemolysis on chemical values for serum. Clin Chem 1978; 24: 1966-70. 3. Burns ER, Yoshikawa N. Hemolysis in serum samples drawn by emergency department personnel versus laboratory phlebotomists. Lab Med 2002; 33: 378-80. 4. AuBuchon JP, Birkmeyer JD, Busch MP. Safety of the blood supply in the United States: Opportunities and controversies. Ann Intern Med 1997; 127: 904-09. 5. Shields CE. Effect of adenine on stored erythrocytes evaluated by autologous and homologous transfusions. Transfusion 1969; 9: 115-19. 6. Monica C. District Laboratory Practice in Tropical Countries, Part 2. Great Britain: Cambridge University Press 2003; 348-61. 7. Afri L, Khan AH, Azeem S. Ionized calcium measurement in serum and plasma by ion selective electrodes: Comparison of measured and calculated parameters. Indian J Clin Biochem 2014; 29: 327-32. 8. Adias TC, Moore-Igwe B, Jeremiah ZA. Storage related haematological and biochemical changes of CPDA-1 whole blood in a resource limited setting. J Blood Disorders Transf 2012; 3: 124. 9. Uvizl R, Klementa B, Adamus M et al. Biochemical changes in the patient’s plasma after red blood cell transfusion. Signa Vitae 2011; 6: 64-71. 10. Hess JR, Sparrow RL, Van der Meer PF et al. Red blood cell hemolysis during blood bank storage: Using national quality management data to answer basic scientific questions. Transfusion 2009; 49: 2599-2603. 11. Heins M, Heil W, Withold W. Storage of serum or whole blood samples? Effects of time and temperature on 22 serum analytes. Eur J Clin Chem Clin Biochem 1995; 33: 231-38. 12. Latham JT Jr, Bove JR, Weirich FL. Chemical and hematologic changes in stored CPDA-1 blood. Transfusion 1982; 22: 158-59. 13. Bailey DN, Bove JR. Chemical and hematological changes in stored CPD blood. Transfusion 1975; 15: 244-49. 14. Koseoglu M, Hur A, Atay A et al. Effects of hemolysis interferences on routine biochemistry parameters. Biochem Med 2011; 21: 79-85. 15. Nsiah K, Dzogbefia VP, Ansong D et al. Pattern of AST and ALT changes in relation to hemolysis in sickle cell disease. Clinical Medicine Insights. Blood Disorders 2011; 4: 1-9. Date of Submission: 17th May 2017 Date of Acceptance: 22nd May. 2017  

Biochemical Changes in Whole Blood - Effect of Storage

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Biochemical Changes in Whole Blood - Effect of Storage

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