Vol 52, No 1 (2021)
Original research article
Published online: 2021-02-26

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Ferric reducing ability of plasma: a potential oxidative stress marker in stored plasma

Carl Hsieh1, Vani Rajashekaraiah1
Acta Haematol Pol 2021;52(1):61-67.


Introduction: The ferric reducing ability of plasma (FRAP) assay is used for measuring the antioxidant capacity. FRAP is proportional to the molar concentration of the antioxidant capacity. This study attempts to analyze the possibilities of FRAP as an indicator of oxidative stress. Methods: Blood was drawn from male Wistar rats and stored for 20 days at 4C in citrate phosphate dextrose adenine 1. The rats were divided into two groups: controls and experimentals. The experimentals were added with antioxidants — l-carnitine, curcumin, vitamin C (VC), and caffeic acid of varying concentrations — 10, 30, and 60 mM (n =5 for each group). Plasma was isolated from these samples at regular intervals (every 5 days), and FRAP and protein were assayed. Results were analyzed by two-way ANOVA, using GraphPad prism 6. FRAP was maintained in controls. Results: VC (ascorbic acid) was the most potent antioxidant in terms of FRAP during storage compared with the above antioxidants. This study emphasizes the use of FRAP as a potential marker of oxidative stress in plasma of stored blood. Conclusion: FRAP can be utilized as a reliable marker for determining the antioxidant capacity.

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  1. Wojdylo A, Oszmianski J, Czemerys R. Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chem. 2007; 105(3): 940–949.
  2. Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. Anal Biochem. 1996; 239(1): 70–76.
  3. Payne AC, Mazzer A, Clarkson GJJ, et al. Antioxidant assays — consistent findings from FRAP and ORAC reveal a negative impact of organic cultivation on antioxidant potential in spinach but not watercress or rocket leaves. Food Sci Nutr. 2013; 1(6): 439–444.
  4. Hisalkar P, Patne A, Karnik A, et al. Ferric reducing ability of plasma with lipid peroxidation in type 2 diabetes. Int J Pharm Biol Sci. 2012; 42: 8–70.
  5. Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem. 2005; 53(10): 4290–4302.
  6. Ndhlala AR, Moyo M, Van Staden J. Natural antioxidants: fascinating or mythical biomolecules? Molecules. 2010; 15(10): 6905–6930.
  7. Racek J, Herynková R, Holecek V, et al. Influence of antioxidants on the quality of stored blood. Vox Sang. 1997; 72(1): 16–19.
  8. Stef DS, Iosif G, Ioan-Trasca T, et al. Evaluation of 33 medicinal plant extracts for the antioxidant capacity and total phenols. J Food Agri Environ. 2010; 8: 207–10.
  9. Cimen MY. Free radical metabolism in human erythrocytes. Clin Chim Acta. 2008; 390(1-2): 1–11.
  10. Desai CT. Antioxidants: fascinating and favourable biomolecules for humans. Science Innovation. 2015; 3(6): 113.
  11. Halliwell B, Gutteridge JM. The definition and measurement of antioxidants in biological systems. Free Radic Biol Med. 1995; 18(1): 125–126.
  12. Carl H, Chandni A, Neha K, et al. Curcumin as a modulator of oxidative stress during storage: a study on plasma. Transfus Apher Sci. 2014; 50(2): 288–293.
  13. Zan T, Tao J, Tang RC, et al. Effect of vitamin C antioxidative protection on human red blood cells. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005; 13: 1106–8.
  14. Ibrahim IH, Sallam SM, Omar H, et al. Oxidative hemolysis of erythrocytes induced by various vitamins. Int J Biomed Sci. 2006; 2(3): 295–298.
  15. Shiva Shankar Reddy CS, Subramanyam MVV, Vani R, et al. In vitro models of oxidative stress in rat erythrocytes: effect of antioxidant supplements. Toxicol In Vitro. 2007; 21(8): 1355–1364.
  16. Mahmud H, Qadri SM, Föller M, et al. Inhibition of suicidal erythrocyte death by vitamin C. Nutrition. 2010; 26(6): 671–676.
  17. Arduini A, Holine S, Sweeney JD, et al. Addition of L-carnitine to additive solution-suspended red cells stored at 4C reduces in vitro hemolysis and improves in vivo viability. Transfus. 1997; 37: 166–74.
  18. Ravikumar S, Rajashekharaiah V. CUPRAC-BCS and antioxidant activity assays as reliable markers of antioxidant capacity in erythrocytes. Hematol. 2015; 20(3): 165–174.
  19. Deyhim MR, Mesbah-Namin SA, Yari F, et al. L-carnitine effectively improves the metabolism and quality of platelet concentrates during storage. Ann Hematol. 2015; 94(4): 671–680.
  20. Vani R, Soumya R, Carl H, et al. Prospects of vitamin C as an additive in plasma of stored blood. Adv Hematol. 2015; 2015: 961049.
  21. Soumya R, Carl H, Vani R. L-carnitine as a potential additive in blood storage solutions: a study on erythrocytes. Indian J Hematol Blood Transfus. 2016; 32(3): 328–334.
  22. Ravikumar S, Hsieh C, Rajashekharaiah V. Prospects of curcumin as an additive in storage solutions: a study on erythrocytes. Turk J Med Sci. 2016; 46(3): 825–833.
  23. Li JL, Wang QY, Luan HY, et al. Effects of L-carnitine against oxidative stress in human hepatocytes: involvement of peroxisome proliferator-activated receptor alpha. J Biomed Sci. 2012; 19: 32.
  24. Meister A. Glutathione-ascorbic acid antioxidant system in animals. J Biol Chem. 1994; 269(13): 9397–9400.
  25. Michels A, Frei B. Vitamin C. In: Caudill MA, Rogers M. ed. Biochemical, physiological, and molecular aspects of human nutrition. Saunders, Philadelphia 2012: 627–654.
  26. Gülçin I. Antioxidant activity of caffeic acid (3,4-dihydroxycinnamic acid). Toxicology. 2006; 217(2-3): 213–220.
  27. Chen J, Ho CT. Antioxidant activities of caffeic acid and its related hydroxycinnamic acid compounds. J Agric Food Chem. 1997; 45(7): 2374–2378.
  28. Olthof MR, Hollman PC, Katan MB. Chlorogenic acid and caffeic acid are absorbed in humans. J Nutr. 2001; 131(1): 66–71.
  29. Godycki-Cwirko M, Krol M, Krol B, et al. Uric acid but not apple polyphenols is responsible for the rise of plasma antioxidant activity after apple juice consumption in healthy subjects. J Am Coll Nutr. 2010; 29(4): 397–406.
  30. Gliszczynska-Swiglo A. Antioxidant activity of water soluble vitamins in the TEAC (trolox equivalent antioxidant capacity) and the FRAP (ferric reducing antioxidant power) assays. Food Chem. 2006; 96(1): 131–136.
  31. Ghasemzadeh A, Jaafar HZE, Rahmat A. Antioxidant activities, total phenolics and flavonoids content in two varieties of Malaysia young ginger (Zingiber officinale Roscoe). Molecules. 2010; 15(6): 4324–4333.
  32. Wang F, Zhao S, Li F, et al. Investigation of antioxidant interactions between Radix Astragali and Cimicifuga foetida and identification of synergistic antioxidant compounds. PLoS One. 2014; 9(1): e87221.
  33. Murugan R, Parimelazhagan T. Comparative evaluation of different extraction methods for antioxidant and anti-inflammatory properties from Osbeckia parvifolia Arn. – An in vitro approach. J King Saud Univ Sci. 2014; 26(4): 267–275.
  34. Rajashekharaiah V, Koshy AA, Koushik AK, et al. The efficacy of erythrocytes isolated from blood stored under blood bank conditions. Transfus Apher Sci. 2012; 47(3): 359–364.
  35. Vani R, Soumya R, Manasa K, et al. Storage lesions in blood components. Antioxid Med Sci. 2015; 4(3): 125.
  36. Dodge JT, Mitchell C, Hanahan DJ. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963; 100: 119–130.
  37. Lowry OH, Rosenberg NJ, Farr AL, et al. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1): 265–275.
  38. Ognik K. Effect of L-carnitine on the level of biochemical and antioxidant indices of blood of turkey hens. Annales UMCS, Zootechnica. 2012; 30(1).
  39. Prabhakar ER, Suchitra MM, Seshadri VR, et al. Ferric reducing ability of plasma and lipid peroxidation in hemodialysis patients: intradialytic changes. Nephro-Urology Monthly. 2010; 2(3): 414–421.
  40. Gülçin I. Antioxidant and antiradical activities of L-carnitine. Life Sci. 2006; 78(8): 803–811.
  41. Surai PF. Antioxidant action of carnitine: molecular mechanisms and practical applications. EC Vet Sci. 2015; 2: 66–84.
  42. Flanagan JL, Simmons PA, Vehige J, et al. Role of carnitine in disease. Nutr Metab (Lond). 2010; 7: 30.
  43. Hsieh C, Rajashekharaiah V. Influence of L-carnitine on stored rat blood: a study on plasma. Turk J Haematol. 2017; 34(4): 328–333.
  44. Barclay LR, Vinqvist MR, Mukai K, et al. On the antioxidant mechanism of curcumin: classical methods are needed to determine antioxidant mechanism and activity. Org Lett. 2000; 2(18): 2841–2843.
  45. Menon VP, Sudheer AR. Antioxidant and anti-inflammatory properties of curcumin. Adv Exp Med Biol. 2007; 595: 105–125.
  46. Barrita JL, Sánchez MD. Antioxidant role of ascorbic acid and his protective effects on chronic diseases and his protective effects on chronic degenerative diseases. Intech, Rejeka 2013: 449-484. https://www.intechopen.com/books/oxidative-stress-and-chronic-degenerative-diseases-a-role-for-antioxidants/antioxidant-role-of-ascorbic-acid-and-his-protective-effects-on-chronic-diseases (May 13, 2021).
  47. Halliwell B. Vitamin C: antioxidant or pro-oxidant in vivo? Free Rad Res. 1996; 25: 439–454.
  48. Elmagirbi A, Sulistyarti H, Atikah A, et al. Study of ascorbic acid as iron(III) reducing agent for spectrophotometric iron speciation. J Pure Appl Chem Res. 2012; 1(1): 11–17.
  49. Son S, Lewis BA. Free radical scavenging and antioxidative activity of caffeic acid amide and ester analogues: structure-activity relationship. J Agric Food Chem. 2002; 50(3): 468–472.