Characterization of erythrocyte quality during the refrigerated storage of whole blood containing di-(2-ethylhexyl) phthalate

Abstract

Di-(2-ethylhexyl) phthalate (DEHP) accumulates in blood brought into contact with materials utilizing this compound as a plasticizer. To determine whether this phthalate diester affects red blood cell integrity, we have compared cell morphology, plasma hemoglobin accumulation, micro-vesicle production, and the concentration of intracellular metabolites and electrolytes of erythrocytes from blood stored at 4 degrees C with and without DEHP. When sufficient emulsified DEHP was mixed with blood to give a final concentration of 300 micrograms/mL, plasma hemoglobin accumulation was reduced by an average of 70%, the percentage of cells exhibiting normal morphology was enhanced by at least 20-fold, and the volume of microvesicles released from red blood cells was reduced by 50% after 35 days of refrigerated storage compared to the values obtained from corresponding samples stored without added phthalate. Similar effects were observed regardless of whether blood was stored in nonplasticized polypropylene or tri-(2-ethylhexyl) trimellitate plasticized polyvinylchloride containers and with DEHP solubilized by a variety of emulsifiers. When 300 micrograms/mL DEHP was added to stored blood containing erythrocytes predominantly in the echinocyte conformation, many of the cells reverted to the normal discoid morphology. The addition of this quantity of DEHP to blood had no significant effect on the course of storage-induced changes in erythrocyte adenosine triphosphate (ATP), 2,3-diphosphoglycerate (2,3-DPG), sodium or potassium concentrations. The data are consistent with the hypothesis that DEHP inhibits the deterioration of the red blood cell membrane that results from the refrigerated storage of whole blood.

2‐(Diethylhexyl)phthalate in blood bags: is this a public health issue?

“…The value of DEHP in RBC storage bags is not only for increased flexibility of the bag but DEHP also improves RBC survival during storage and after transfusion 17 . DEHP affects RBC integrity by inhibiting the deterioration of the RBC membrane, which prevents hemolysis, microvesicular formation, and shape change that occur during refrigerated storage 18 . The potential mechanism for the increased RBC stability is because DEHP is associated with the RBC membrane and cystosol 2 .…”

Storage of red blood cells in a novel polyolefin blood container: a pilot in vitro study

“…Storage of RBC in blood bags manufactured of plastic materials that do not include plasticizers will be associated with new problems, different from what we generally experience in RBC storage studies. Plasticizers, in particular DEHP, have a stabilizing effect on the erythrocyte membrane as the plasticizer will be incorporated into the interior and membrane fractions of erythrocytes and thereby reduce osmotic fragility and in a second step haemolysis . For this reason, we started this evaluation of a new blood bag system manufactured of plastic materials without plasticizers with RBC storage studies with a special focus on haemolysis.…”

“…Not until in the 1970s, the migration of DEHP from the plastic into blood components was fully recognized . Ten years later, plasticizer effects on erythrocyte membranes were reported . The situation may briefly be summarized in the following way, (1) DEHP migrates into stored blood components due to its lipophilic characteristics, implicating that the presence and levels of plasma lipids will increase migration; (2) for this reason, the quantity of DEHP extracted may vary between blood from different donors; (3) DEHP has a stabilizing effect on the erythrocyte membrane as DEHP is incorporated into the interior and membrane fractions of erythrocytes thus reducing osmotic fragility and thereby haemolysis; (4) the migration of DEHP is temperature dependent, increasing at higher temperature and (5) at a fixed temperature, the migration of DEHP is essentially linear over time.…”

Biochemical and structural changes in RBCs stored with different plasticizers: the role of hexanol

“…The 3 m M concentration was calculated to result in 144.6 μg per mL in our preparations. The values reported as present or added by other investigators ranged from 146 to 270 μg per mL 17,19,39 …”